Engineered Nanoparticles For Drug Delivery in Cancer

Angewandte
. Y. Xia et al.
Reviews
DOI: 10.1002/anie.201403036
Nanomedicine
Engineered Nanoparticles for Drug Delivery in Cancer

Therapy
Tianmeng Sun, Yu Shrike Zhang, Bo Pang, Dong Choon Hyun, Miaoxin Yang,
and Younan Xia*
Keywords:
cancer therapy · controlled release ·
drug delivery · nanomedicine ·
nanoparticles
Angewandte
Chemie
&&&& 2014 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim Angew. Chem. Int. Ed. 2014, 53, 2 – 47
Ü
Ü
These are not the final page numbers!

Angewandte
Nanoparticular Drug Delivery Chemie
In medicine, nanotechnology has sparked a rapidly growing interest as From the Contents
it promises to solve a number of issues associated with conventional
1. Introduction 3
therapeutic agents, including their poor water solubility (at least, for
most anticancer drugs), lack of targeting capability, nonspecific 2. Working with Different Types of
distribution, systemic toxicity, and low therapeutic index. Over the past Anticancer Drugs 5
several decades, remarkable progress has been made in the develop-
ment and application of engineered nanoparticles to treat cancer more 3. Methods for Controlled Release 8
effectively. For example, therapeutic agents have been integrated with 4. In vitro and in vivo Delivery 12
nanoparticles engineered with optimal sizes, shapes, and surface
properties to increase their solubility, prolong their circulation half- 5. Perspectives on the Design of
life, improve their biodistribution, and reduce their immunogenicity. Nanoparticle Carriers 24
Nanoparticles and their payloads have also been favorably delivered
6. Case Studies 27
into tumors by taking advantage of the pathophysiological conditions,
such as the enhanced permeability and retention effect, and the spatial 7. Summary and Outlook 36
variations in the pH value. Additionally, targeting ligands (e.g., small
organic molecules, peptides, antibodies, and nucleic acids) have been
added to the surface of nanoparticles to specifically target cancerous Nanomedicine, the application of
cells through selective binding to the receptors overexpressed on their nanotechnology to medicine, is antici-
surface. Furthermore, it has been demonstrated that multiple types of pated to help us move toward the
therapeutic drugs and/or diagnostic agents (e.g., contrast agents) could aforementioned goals. After several
decades of technological develop-
be delivered through the same carrier to enable combination therapy
ments, drug-delivery systems based
with a potential to overcome multidrug resistance, and real-time on engineered nanoparticles have
readout on the treatment efficacy. It is anticipated that precisely engi- started to show great promise.[7] As
neered nanoparticles will emerge as the next-generation platform for shown in Figure 1, the nanoparticles
cancer therapy and many other biomedical applications. used for drug delivery can be readily
fabricated from either soft (organic
and polymeric) or hard (inorganic)
materials, with their sizes being con-
1. Introduction trolled typically in the range of 1–100 nm and compositions/
structures being engineered to load anticancer drugs in
Cancer is one of the leading causes of death, accounting a variety of configurations.[8] The physicochemical properties
for 8.2 million deaths worldwide in 2012.[1] Over the past of the nanoparticles can also be finely tuned by tailoring their
several decades, remarkable breakthroughs have been made chemical compositions, sizes, shapes, structures, morpholo-
in advancing our understanding of how cancer originates and gies, and surface properties.[2, 7a,e, 9] A number of such delivery
develops, which has in turn led to better methods for both systems have been approved for cancer therapy in the clinics,
diagnosis and treatment.[2] Although the overall mortality of with many more currently under clinical trials or preclinical
cancer is showing a declining trend for the first time in five evaluations (see Table 1 for a list). Nanoparticle-based
decades, it still remains at a high rate of 20.2 %.[3] In the therapeutics are poised to significantly improve the treatment
United States alone, for example, cancer resulted in an outcomes for oncological diseases, promising to reshape the
estimated 580 350 deaths out of 1 660 290 total diagnoses in landscape of the pharmaceutical industry.[9]
2013.[4] A major reason for this high mortality rate lies in our Compared with traditional chemotherapeutics, the deliv-
inability to deliver therapeutic agents only to the tumor sites ery of anticancer drugs through a nanoparticle-based plat-
without inducing severe adverse effects on healthy tissues and
organs.[5] In addition to surgical intervention, current cancer
treatments heavily rely on radiation and chemotherapeutic [*] Dr. T. Sun,[+] Dr. Y. S. Zhang,[+] B. Pang,[+] Dr. D. C. Hyun,[+] Prof. Y. Xia
agents, which also kill “normal” cells and cause toxicity to the The Wallace H. Coulter Department of Biomedical Engineering,
patient. Therefore, it would be desirable to develop highly Georgia Institute of Technology and Emory University
Atlanta, GA 30332 (USA)
efficient therapeutics, the so-called “magic bullets”, that can
M. Yang, Prof. Y. Xia
overcome biological barriers, distinguish between malignant
School of Chemistry and Biochemistry
and benign cells, selectively target the cancerous tissues, and Georgia Institute of Technology
“intelligently” respond to the heterogeneous and complex Atlanta, GA 30332 (USA)
microenvironment inside a tumor for on-demand release of E-mail: [email protected]
therapeutic agents in the optimal dosage range.[6] [+] These authors contributed equally to the preparation of this review
article.
Angew. Chem. Int. Ed. 2014, 53, 2 – 47 2014 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim www.angewandte.org &&&&
Ü
Ü

.
Angewandte Y. Xia et al.
Reviews
Figure 1. A summary of nanoparticles that have been explored as carriers for drug delivery in cancer therapy, together with illustrations of
biophysicochemical properties.
form offers many attractive features, including: 1) improved the highly acidic environment in the stomach or the lysosomes
delivery of drugs that are poorly soluble in water and delivery of a cell, and the high levels of proteases or other enzymes in
of a therapeutic agent into cancerous cells at a high dose; the blood stream) before they can reach the targets, leading to
2) better protection of a drug from harsh environments (e.g., an extended plasma half-life of the drug in the systemic
Younan Xia studied at the University of Yu Shrike Zhang received his B.S. in Bio-
Science and Technology of China (B.S. in medical Engineering from Southeast Univer-
1987) and UPenn (M.S. in 1993), and sity, China, in 2008. He received his Ph.D.
received his Ph.D. from Harvard in 1996 in biomedical engineering (with Prof.
(with George Whitesides). He started as Younan Xia) in 2013, and then joined Prof.
Assistant Professor at the University of Wash- Ali Khademhosseini’s group as a postdoctoral
ington (Seattle) in 1997, and was promoted fellow at Harvard Medical School, Brigham
to Associated Professor and Professor in and Women’s Hospital, and Harvard-MIT
2002 and 2004, respectively. He joined the Division of Health Sciences and Technology.
Department of Biomedical Engineering at His research interests include biomaterials,
Washington University in St. Louis in 2007 tissue engineering, regenerative medicine,
as the James M. McKelvey Professor. In early imaging, nanomedicine, and lab-on-a-chip.
2012, he moved to Georgia Tech to take the
position of Brock Family Chair and GRA Eminent Scholar in Nano-
medicine. His research interests include nanomaterials, biomaterials, nano-
medicine, and regenerative medicine.
Tianmeng Sun has been a postdoctoral Bo Pang received his B.E. in mechanical
fellow in the Xia group at Georgia Tech engineering (2010) and B.S. in chemistry
since August 2012. He received both his (2011) from Peking University, China. Cur-
B.S. (in 2006) and Ph.D. (in 2011) in life rently, he is pursuing his Ph.D. degree in
science and cell biology from the University biomedical engineering in the joint Ph.D.
of Science and Technology of China. His program of Georgia Tech/Emory/Peking
thesis focused on the development of nano- University under the supervision of Profs.
particular delivery systems for cancer therapy Qiushi Ren and Younan Xia. His research
based on siRNAs and chemotherapeutic interests focus on the use of gold nanocages
agents. His current research interests focus for cancer therapy and nanoparticle-based
on the development of nanomaterials for imaging contrast agent for PET and SPECT.
cancer and atherosclerosis treatments.
&&&& www.angewandte.org 2014 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim Angew. Chem. Int. Ed. 2014, 53, 2 – 47
Ü
Ü

Angewandte
circulation; 3) targeted delivery of drugs in a cell- or tissue- 2. Working with Different Types of Anticancer Drugs
specific manner so the treatment efficacy can be maximized
while systemic side effects are alleviated; 4) controlled Broadly speaking, cancer therapeutic agents can be
release of drugs over a manageable period of time at precise classified into two major groups, hydrophobic and hydro-
doses and even realization of on-demand release using a more philic, depending on their aqueous solubility. Alternatively,
sophisticated, stimuli-responsive system; and 5) co-delivery they can be categorized as highly charged or neutral drugs
of multiple types of drugs and/or diagnostic agents (e.g., based on their electrostatic properties (Table 2). When
contrast agents) for combination therapy (which has the choosing or designing nanoparticles to be used as the carrier
potential to overcome multidrug resistance) and real-time for a specific type of drug, it is of critical importance to know
readout on the treatment efficacy.[7h, 37] the properties and behaviors of the drug in order to achieve
In this Review, we first discuss the critical need for an optimal encapsulation efficiency and the desired release
nanoparticle carriers for the delivery of various types (hydro- profile. In this section, we use a set of examples to highlight
philic, hydrophobic, and highly charged) of cancer therapeu- the challenges faced by the application of various anticancer
tic agents. We then present a number of strategies based on drugs and then discuss strategies that can potentially over-
diffusion, erosion, and stimuli-responsive triggering for reg- come these obstacles by using engineered nanoparticles as the
ulating the release of drugs from nanoparticle carriers. carriers.
Following that, we highlight some general challenges in
targeted delivery of nanoparticle carriers under in vitro and
in vivo conditions, including endocytosis, intracellular trans- 2.1. Hydrophobic Drugs
port, biodistribution, biocompatibility, biodegradability, and
systemic clearance. We then elaborate on the design, syn- The majority of anticancer drugs currently used in the
thesis/fabrication, and functionalization of nanoparticle car- clinic are hydrophobic, including, for example, paclitaxel,
riers based on the requirements imposed by various applica- which is widely used for treating ovarian, breast, and non-
tions. Finally, we illustrate how the concept of nanomedicine small-cell lung cancers. How to effectively deliver a hydro-
has been materialized by focusing on some selected examples phobic drug to its target has always been a challenge and
of nanoparticle carriers, including protein conjugates, lip- a subject of active research. The reason lies in the fact that
osomes, dendrimers, as well as those composed of organic hydrophobic drug molecules may not be soluble enough to
polymers, hydrogels, phase-change materials, and inorganic cross the aqueous environment (e.g., the body and tissue
materials. fluids in vivo) surrounding a cell and then penetrate the cell
membrane to eventually reach intracellular targets. Addi-
tionally, their strong tendency to aggregate upon intravenous
administration can lead to complications such as embolisms
Dong Choon Hyun has been a postdoctoral
and local toxicity.[51]
fellow in the Xia group at Georgia Tech One efficient approach to overcome the poor water
since October 2012. He received his B.S. (in solubility of a hydrophobic drug is to encapsulate it in
2006) and Ph.D. (in 2011) degrees in a nanoparticle-based carrier. The primary requirement for the
materials science and engineering from carrier is a good loading capacity for the hydrophobic drug,
Yonsei University, Korea. His thesis focused which can be easily met through the use of a hydrophobic or
on the fabrication and application of poly-
amphiphilic material.[37a] A variety of carriers have thus been
mer micro- and nanostructures. His current
research interests include the design and developed for the delivery of hydrophobic drugs, including
fabrication of biodegradable polymer parti- those based on polymer micelles and polymer nanoparti-
cles and exploration of their applications in cles.[2, 52] For example, Allen and co-workers demonstrated
drug delivery. that the concentration of ML220 (a highly hydrophobic aryl-
imidazole drug) in an aqueous medium could be increased by
a factor of more than 50 000 when it was encapsulated in
a liposome-based carrier.[53] Park and co-workers found that
nanosized micelles based on amphiphilic block copolymers
could serve as a carrier for the delivery of drugs poorly soluble
Miaoxin Yang received his B.S. in pharmacy in water (such as paclitaxel), significantly increasing the drug
from Shanghai Jiaotong University in 2006,
concentration in an aqueous medium by a factor of more than
M.S. in chemistry from Nanyang Technolog-
ical University in 2009, and M.S. in biomed- 1000.[54] Similar improvement has also been observed with
ical engineering from Washington University solid nanoparticles made of biocompatible, biodegradable
in St. Louis in 2011. After working for polymers.[55]
Philips as a research scientist for two years,
he joined the Xia group in 2013 to pursue
a Ph.D. degree in chemistry and biochem-
2.2. Hydrophilic Drugs
istry at Georgia Tech. His research interests
include the development of novel nanomate-
rials for biomedical applications. Hydrophilic drugs, including biomacromolecules (e.g.,
proteins, peptides, and nucleic acids)[56] and many small
Ü
Ü

.
Reviews
Table 1: Nanoparticle-based therapeutics in clinical use and under clinical investigation.

Trade name Formulation Drug Company Application Phase of
development
Abraxane albumin-bound nanoparti- paclitaxel Abraxis Bioscience, metastatic breast cancer[10] approved
cle Inc.
Caelyx PEGylated liposome doxorubicin Schering-Plough metastatic breast and ovarian cancer, approved
Kaposi sarcoma[11]
DaunoXome liposome daunorubicin Galen Ltd Kaposi sarcoma[12] approved
DepoCyt lipodome cytarabine Pacira Pharmaceuti- lymphoma[13] approved
cals, Inc.
Doxil liposome doxorubicin Sequus Pharmaceuti- Kaposi sarcoma[14] approved
cals, Inc.
Genexol-PM polymeric micellar nano- paclitaxel Samyang Biopharma- breast cancer[15] approved
particle ceuticals
Marqibo liposome vincristine sulfate Talon Therapeutics, lymphoblastic leukemia[16] approved
Inc.
Myocet liposome doxorubicin Zeneus Pharma Ltd metastatic breast cancer[17] approved
Oncaspar PEGylated asparaginase asparaginase Enzon Pharmaceuti- acute lymphoblastic leukemia[18] approved
cals, Inc.
Zinostatin stima- poly(styrene-co-maleic neocarzinostatin Astellas Pharma, Inc. hepatocellular carcinoma[19] approved
lamer acid)-conjugated neocarzi-
nostatin
NK105 micellar nanoparticle paclitaxel Nippon Kayaku Co., breast cancer[20] phase III
Ltd
BIND-014 polymer matrix docetaxel BIND Therapeutics, prostate cancer[21] phase II
Inc.
Genexol-PM methoxy PEG-PLA paclitaxel Samyang Biopharma- ovarian and lung cancer[22] phase II
ceuticals
CRLX101 cylodextrin-PEG micelle camptothecin Cerulean Pharma, ovarian/tubal/peritoneal cancer, rectal phase I/II
Inc. cancer[23]
CYT-6091 gold nanoparticle tumor necrosis Cytimmune Sciences, pancreatic cancer, melanoma, soft- phase I/II
factor a Inc. tissue sarcoma, ovarian, and breast
cancer[24]
l-Annamycin liposome annamycin Callisto Pharmaceuti- acute lymphocytic leukemia, acute phase I/II
cals, Inc. myelogenous leukemia[25]
NL CPT-11 liposome irinotecan University of Califor- solid tumor[26] phase I/II
nia, San Francisco
Rexin-G pathotropic nanoparticle dominant negative Epeius Biotechnolo- breast cancer, osteosarcoma[27] phase I/II
cyclin G1 construct gies
Anti-EGFR immu- liposome doxorubicin University Hospital, solid tumor[28] phase I
noliposome Switzerland
AuroLase gold nanoparticle Nanospectra Biosci- lung cancer, head and neck cancer[29] phase I
ences, Inc.
BikDD nanoparti- liposome proapoptotic Bik National Cancer pancreatic cancer[30] phase I
cle gene (BikDD) Institute
CALAA-01 cyclodextrin-containing siRNA Calando Pharmaceut- solid tumor[31] phase I
polymer icals, Inc.
CRLX301 cyclodextrin-based polymer docetaxel Cerulean Pharma, solid tumor[32] phase I
Inc.
DEPTM-Docetaxel dendrimer docetaxel Starpharma Hold- breast, prostate, lung, and ovarian phase I
ings, Ltd cancer[33]
Docetaxel-PNP polymeric nanoparticle docetaxel Samyang Biopharma- advanced solid malignancies[34] phase I
ceuticals
TKM-080301 lipid nanoparticle siRNA National Institutes of liver cancer[35] phase I
Health Clinical
Center
C-dots PEG-coated SiO2 C-dots Development melanoma[36] IND
approved
molecules,[57] also play an important role in treating various gemcitabine, a nucleoside analogue, has been used to treat
types of cancers. For example, trastuzumab, a monoclonal bladder, pancreas, ovarian, breast, and non-small-cell lung
antibody that interferes with the human epidermal growth cancers. Nevertheless, successful utilization of hydrophilic
factor receptor 2 (HER2), is now routinely used to treat early- drugs has been hindered by a number of obstacles, such as
stage and metastatic breast cancer for many years; and poor uptake by cells because of their inability to cross the
Ü
Ü

Angewandte
Table 2: Anticancer drugs currently in clinical use.[a]

Drug Clinical use
Hydrophobic
adriamycin ALL, AML, Wilms’ tumor, neuroblastoma, soft-tissue and bone sarcomas, breast, ovary, urinary bladder, thyroid, gastric,
Hodgkin’s disease[38]
cisplatin metastatic testicular tumors, metastatic breast cancer, glioblastoma, lung, lymphoma, anal canal[38]
docetaxel breast, NSCLC, HRPC, gastric adenocarcinoma, SCCHN[38]
etoposide SCLC, GCT, lymphoma, Ewing’s sarcoma, ovary[39]
methotrexate gestational choriocarcinoma, ALL, breast, epidermoid cancer of the head and neck, advanced mycosis[38]
paclitaxel advanced carcinoma of the ovary, breast, lung, stomach, head and neck, GCT, urinary bladder, urothelial[40]
Hydrophilic
bevacizumab metastatic CRC, NSCLC, Metastatic breast cancer, glioblastoma, ovary, renal cell carcinoma[41]
cetuximab metastatic colorectal and non-small-cell lung carcinoma[42]
cyclophosphamide malignant lymphomas, Hodgkin’s disease, lymphocytic lymphoma, CLL, CML, ALL, AML, osteosarcoma, GCT[43]
gemcitabine breast, NSCLC, pancreas, ovary, gallbladder, lymphoma, urinary bladder[44]
ibritumomab B-cell non-Hodgkin’s lymphoma[45]
l-asparaginase ALL[46]
panitumumab metastatic colorectal carcinoma[47]
rituximab CD20-positive B-cell non-Hodgkin’s lymphoma[48]
tositumomab CD20-positive B-cell non-Hodgkin’s lymphoma[49]
transtuzumab metastatic breast cancer[50]
Highly charged
DNA no clinical use
siRNA/miRNA no clinical use
[a] ALL (acute lymphoblastic leukemia), AML (acute myeloid leukemia), CLL (chronic lymphocytic leukemia), CML (chronic myelogenous leukemia),
CRC (colorectal cancer), GCT (granulosa cell tumor), HRPC (hormone refractory prostate cancer), NHL (hon-Hodgkin lymphoma), NSCLC (non-
small-cell lung cancer), SCCHN (squamous cell carcinoma of the head and neck), SCLC (small-cell lung cancer).
lipid-rich, hydrophobic cell membranes, low bioavailability phase.[62] For some of these modifications, the poor dispersion
arising from their poor stability against proteolytic and of a hydrophilic drug in nanoparticles, which often results in
hydrolytic degradation, and short half-life in the circulatory rapid release of the drug, can also be largely addressed.[63]
system.[56, 58]
To circumvent these hurdles, nanoparticles have been
actively explored as carriers to encapsulate and deliver 2.3. Highly Charged Drugs
hydrophilic drugs. Similar to hydrophobic drugs, loading
efficiency is also one of the major issues to consider because Gene therapeutics based on DNA, siRNA, and micro-
the overall dosage has to be increased when nanoparticles RNA represent a special class of hydrophilic drugs with high
with low drug contents are administered.[59] Considering the densities of charges. They were developed over the past few
hydrophobic nature of most materials used for fabricating the decades and are expected to serve as a powerful molecular
carriers, loading of a hydrophilic drug into such a delivery therapy for the treatment of various diseases.[64] These drugs
system is not always straightforward, owing to the poor have all the characteristics of a hydrophilic drug, such as poor
miscibility between these two phases. To this end, a number of cellular uptake and fast degradation in the physiological
approaches have been developed to improve the loading milieu. Additionally, systemic administration of these ther-
efficiency of a hydrophilic drug. For instance, Hall and co- apeutics is impeded by barriers such as rapid clearance by the
workers replaced 5-fluorouracil (5-FU, a hydrophilic pyrimi- mononuclear phagocyte system (MPS) and kidney filtration.
dine analogue for cancer treatment) with 1-alkylcarbonylox- It should be pointed out that there has been no clinical success
ymethyl (an amphiphilic prodrug of 5-FU) to significantly in gene therapy to date. In a sense, efficient delivery of these
increase the drug loading efficiency from 3.68 % to 47.23 %.[60] highly charged agents through proper carriers remains
Fattal and co-workers discovered that adjusting the pH value a major obstacle for achieving their therapeutic benefits.[65]
of the external aqueous phase to the isoelectric point of Currently, loading and delivery of gene drugs rely on the
a protein drug could increase the drug loading.[61] Xu and co- electrostatic interactions between the highly charged nucleic
workers demonstrated that the electrostatic and hydrophobic acids and nanoparticle carriers. As the nucleic acids are
interactions between lipidoids and a protein drug could be usually negatively charged under physiological conditions, it
enhanced to facilitate the formation of protein–lipidoid is reasonable to rely on the use of positively charged carriers,
complexes and thus intracellular delivery.[59c] Furthermore, such as liposomes and polymer nanoparticles consisting of
McGinity and co-workers found that a less hydrophilic cationic building blocks, to improve the loading efficiency.[66]
organic solvent in the oil phase could prevent the encapsu- To this end, Wang and co-workers developed a positively
lated hydrophilic drugs from releasing into the outer water charged micelle system composed of amphiphilic and cationic
Ü
Ü

.
Reviews
triblock copolymers, which could effectively immobilize

siRNA for intracellular delivery and release.[67] Unfortu-
nately, some of these nanoparticles can cause severe problems
associated with toxicity and immune or inflammatory
responses,[68] which need to be taken into consideration
when designing the next-generation carriers. Direct conjuga-
tion of a gene drug to the surface of nanoparticle carrier has
been demonstrated as another effective approach.[69] By
immobilizing nucleic acids on the surface, the complexity of
a loading process may be reduced relative to the encapsula-
tion method. In one example, Mirkin and co-workers initially
coated Au nanoparticles with a layer of short DNA chains
containing specially designed sequences. Using these DNA-
coated Au nanoparticles as the carrier, they were able to
further immobilize a target DNA or siRNA of interest for
efficient delivery of the therapeutics.[69]
3. Methods for Controlled Release Figure 2. Illustration of three major mechanisms for achieving sus-
tained drug release: a,b) diffusion through an insoluble polymer shell
Placing drug molecules inside or on the surface of or matrix, and c) erosion of a polymer matrix. Reproduced with
permission from Ref. [71b], copyright 2010 Springer.
a nanoparticle carrier allows for controlled release, which
offers multiple benefits compared to the conventional dosing
forms based on free drugs. For example, it can improve the 3.1.1. Diffusion-Controlled Release
temporal and spatial presentations of a drug in the body,
protect the drug from physiological degradation or elimina- In diffusion-controlled release, the drug molecules pre-
tion, reduce toxicity to the healthy tissues and organs, and loaded inside a nanoparticle are restricted from entering the
increase patient compliance and convenience. Great strides aqueous environment by a barrier provided by an insoluble
have been made in the design and development of nano- material (typically, an organic polymer). In general, diffusion-
particle-based systems for controlled release,[70] and their controlled release can be realized using a reservoir- or matrix-
operation modes can be broadly classified into two major based system.[73] By its name, a reservoir-based system
categories: sustained and stimuli-responsive (i.e., smart) consists of a core reservoir that contains the drug and
release. Here we only discuss the general principles of a membrane surrounding the reservoir (Figure 2 a). The
controlled release that can be readily applied to nanoparti- concept of this release system can be easily extended to the
cle-based carriers. nanosized carriers by switching to colloidal hollow parti-
cles.[74] Typically, the drug molecules initially loaded in the
reservoir can only diffuse out through the membrane. The
3.1. Sustained Release drug release rate is determined by the physicochemical
properties of the membrane and the loaded drug, as well as
Sustained release aims to deliver a drug at a predetermined the thickness of the membrane. In spite of their simplicity in
rate over an extended period of time. This mode of release is release mechanism and their capability to achieve a steady-
critical for drugs that are rapidly metabolized and eliminated state release, polymer-membrane-based systems have a crit-
from the body after administration. The sustained release can ical drawback because of the undesired dose from pinholes
maintain the concentration of the drug at a constant level in and cracks that may form in the membrane.[75] Recently,
the plasma or target tissue by matching the rate of drug hollow nanoparticles made of inorganic materials have been
release with the rate of drug elimination. In the case of cancer developed for drug delivery in cancer therapy.[76] For example,
therapy, maintaining the concentration of a drug within the Fe3O4 hollow nanoparticles with pores of around 3 nm in size
therapeutic window is beneficial to the patient. could release cisplatin through the pores by a diffusion-
Once dissolved in the aqueous body fluid, most drugs can controlled, slow process. The porous shell was mechanically
be freely transported with the fluid to quickly reach the target and physiologically stable.[77]
receptors. One approach to achieve sustained release is to In matrix-based systems (Figure 2 b), which have been
prevent the drug molecules from entering the aqueous most extensively explored, the drug molecules are uniformly
environment for a controllable period of time. As shown in dispersed in the carrier made of a water-insoluble polymer,
Figure 2, the prevention can be realized by controlling the such as polyurethane (PUA) and poly(methyl methacrylate)
diffusion of drug molecules through an insoluble polymer (PMMA). These systems typically display a noticeable release
shell or matrix, or by simply controlling the degradation rate at the initial point (i.e., the so-called burst release) owing to
of a carrier.[71] The release mechanisms and the corresponding the desorption of drug molecules adsorbed on the surface of
mathematical models have been extensively studied and the nanoparticles.[78] In the following steps, the release will be
reviewed.[72] retarded because it takes time for the drug molecules inside
Ü
Ü

Angewandte
the polymer matrix to diffuse to the surface. This retardation intermediate rate when the polymer gradually degrades. In
effect is more prominent for spherical carriers, as the number the last stage, the trapped drug is rapidly released upon
of available drug molecules decreases with the distance from complete destruction of the polymer matrix.
the surface.[71b, 75] In one study, it was shown that PUA In practice, the release of drug molecules from nano-
nanoparticles only released 40 % of the preloaded doxorubi- particles made of an erodible polymer is much more complex
cin during six days of incubation.[79] Similarly, only 35 % of the than those based on diffusion because various release
encapsulated docetaxel (Dtxl) was released from the PCL- mechanisms can be simultaneously involved. For nanoparticle
Tween 80 nanoparticles over a period of 28 days.[80] formulated with large specific surface areas, bulk and surface
erosions have been shown to occur concurrently. A further
3.1.2. Erosion-Controlled Release discussion on this system in cancer therapy can be found in
Section 6.3.
Nanoparticle carriers made of erodible or degradable
polymers (Figure 2 c) have attracted much attention in recent
years because they do not require retrieval or further 3.2. Stimuli-Responsive Release
manipulation after the drug is fully released.[80] The pattern
of drug release can be controlled by tailoring the erosion There are many clinical situations that require treatments
kinetics of nanoparticles through careful selection of poly- beyond sustained, continuous release of drugs.[87] Studies in
mers and encapsulation techniques.[81] chronopharmacology indicate that the onsets of certain
A number of biodegradable polymers, both synthetic and diseases, such as tumorigenesis and progression of cancer,
natural, have been used for formulating erodible polymer exhibit strong circadian dependence.[88] Treatment of such
nanoparticles. Synthetic polymers have the advantage of diseases requires smart control over the drug release patterns
sustained release of preloaded drugs over periods of days to and profiles in response to in vivo physiological conditions or
several weeks. Representative examples include polyesters external stimuli.[89] With respect to the biological system, the
such as poly(lactic acid) (PLA), poly(glycolic acid) (PGA), stimuli used to trigger the release of a drug can be broadly
and poly(lactic-co-glycolic acid) (PLGA). These polymers are classified as either internal (e.g., variation in pH value or
degraded through hydrolytic cleavage of the ester bond concentrations of ions, small molecules, and enzymes)[70e, 90] or
between lactic and glycolic acid, and thus can be easily external (e.g., light, ultrasound, electric field, magnetic field,
metabolized in the body and eliminated as carbon dioxide and and heating, which are also commonly referred to as physical
water.[82] During the hydrolytic process, the accessibility of stimuli).[70e, 90d, 91] In principle, the fast response of a drug-
water molecules to polymer matrices (i.e., the hydrophilicity delivery system to a stimulus can be employed for real-time
of polymers) determines the erosion rate. Furthermore, the manipulation of drug dosage and further achievement of on-
hydrolysis is dependent on the local concentrations of proton demand drug delivery. A wide spectrum of stimuli-responsive
donors and acceptors. As the degraded monomers of certain materials and their detailed mechanisms in controlling release
polymers (e.g., PLA, PGA, and PLGA) provide acidic have been extensively reviewed elsewhere.[70d, 92] In the
protons, their degradation rate may be self-expedited upon following sections, we only focus on several representative
accumulation of these acidic products.83] The addition of stimuli-responsive release systems used in cancer therapy.
external acidic or basic excipients can also regulate the rate of
polymer erosion. 3.2.1. pH-Sensitive Release
Polymer erosion can proceed through a surface or bulk
mechanism.[84] Surface erosion takes place when the rate of The variation in pH value associated with a pathological
erosion is faster than the rate of water permeation into the situation such as cancer or inflammation has been extensively
bulk of the polymer. This is considered to be a preferred used to trigger the release of a drug into a specific organ (e.g.,
mechanism of erosion for drug delivery because a steady-state gastrointestinal tract or vagina) or intracellular compartment
release of the drug can be reproducibly achieved using a very (e.g., endosome or lysosome).[70e] Many anticancer drug-
thin polymer or by keeping the surface area of the carrier delivery systems have exploited the difference in pH values
constant. One good example of surface-eroding polymers is existing between healthy tissues (ca. 7.4) and the extracellular
polyanhydride,[85] whose exceptional hydrophobicity retards environment of solid tumors (6.5–6.8). One approach is to use
water permeation while its highly labile groups lead to rapid polymers with functional groups that can alter the density of
hydrolysis when encountering water molecules. charges in response to pH variation as the nanoparticle
Bulk erosion occurs if water molecules can imbibe into the carriers. Notable examples include poly(acryl amide)
polymer more rapidly than erosion takes place.[71] In this case, (PAAm), poly(acrylic acid) (PAA), poly(methacrylic acid)
chain scission occurs throughout the matrix, leading to a very (PMAA), poly(methyl acrylate) (PMA), poly(diethylami-
complex degradation/erosion process for the polymer. Poly- noethyl methacrylate) (PDEAEMA), and poly(dimethylami-
esters, the most commonly used biodegradable polymers for noethyl methacrylate) (PDMAEMA). With the use of these
controlled release, work by bulk erosion. The release of drug polymers, the structure and hydrophobicity of the nano-
from a bulk-eroding polymer typically undergoes three particle carriers can change as a result of protonation or
stages.[86] In the first stage, drug is released from the surface deprotonation.[93]
or from pores that are connected to the surface. During the Figure 3 schematically illustrates how the polymer chains
second stage, the remaining drug is released at a slow to either extend or collapse in response to the variation in
Ü
Ü

.
Reviews
end, Rhov and co-workers syn-

thesized N-(2-hydroxypropyl)
methacrylamide (HPMA), a poly-
mer conjugated with hydrazone
groups for the attachment of dox-
orubicin. The drug was released
from the conjugates at pH 5.[99]
Similarly, doxorubicin conjugated
to a hydrazone-linked dendrimer
was released in a pH-sensitive
manner. The release was rapidly
completed at pH 5, reaching 100 %
release within 48 h.[100] Kratz and
co-workers developed an acid-
cleavable doxorubicin prodrug
Figure 3. Schematic illustration of drug release from a polymer micelle containing polycarboxylates, derived from dendritic polygly-
a liposome, and a hydrogel nanosphere initiated by a variation in the pH value. a) Protonation (left)
cerol. Use of the hydrazone linker
or deprotonation (right) results in destruction of the polymer micelle. b) Protonation induces collapse
led to a dramatic change in drug
of the polyanion chains, making the liposomal shell leaky and thus promoting efflux of the drug from
the liposome. c) Deprotonation leads to swelling of the hydrogel matrix, triggering drug release from release between pH 5 and 6.[101] A
the nanosphere. Reproduced with permission from Ref. [98a], copyright 2012 Elsevier. polymer containing an acid-degrad-
pH value and thereby cause changes to the

electrostatic interactions, allowing for pH-
dependent release of drug molecules.[93d,e]
For example, micelles derived from block
co-polymers can release the preloaded drug
when the pH value is varied (Figure 3 a).
One way to trigger the release is to reduce
electrostatic interactions between a posi-
tively charged drug (such as doxorubicin)
and an oppositely charged block copolymer
by protonating the carboxylate groups of the
block copolymer.[94] Polymer chains sensitive
to the pH value can also be incorporated into
liposome to make the permeability of the
phospholipid bilayers dependent on
pH value (Figure 3 b).[93e] For example, the
protonation of poly-2-vinylpyridine (P2VP)
blocks at pH 4.9 induced rupture of the
membrane of PEG-b-P2VP polymersomes, Figure 4. Cleavable linkers that have been used for stimuli-responsive drug release. The
triggering the release of the encapsulated dotted line in each molecule indicates the bond that will be broken upon activation by the
corresponding stimulus (indicated in parentheses). Reproduced with permission from
drug.[95] Hydrogel nanoparticles can be
Ref. [70d], copyright 2012 Elsevier.
employed for pH-sensitive release as well,
where changes to the degree of protonation/
deprotonation lead to swelling and/or shrinking, triggering able backbone was obtained through terpolymerization of
the release of drug molecules (Figure 3 c).[96] For example, PEG, divinyl ether, and serinol. The pendant amino groups
Bae and co-workers demonstrated the synthesis of a self- can be used for the conjugation of doxorubicin to the polymer
assembled, pH-responsive hydrogel. When loaded with backbone.[102] In another demonstration, acid-degradable
doxorubicin, this system showed an enhancement in toxicity linkers such as the pH-labile hydrazone bond was exploited
at pH 6.8, which is similar to the pH level inside the tumor to trigger the release of a drug. Such a linker allows the
tissue.[97] Another approach is to incorporate cleavable bonds attachment of doxorubicin to the hydrophobic block of an
into the nanoparticle carrier. The cleavable bonds can be amphiphilic polymer.[103]
broken to directly release the drug molecules conjugated to or
encapsulated in the carrier.[69e, 98] Figure 4 shows a partial list 3.2.2. Enzyme-Sensitive Release
of cleavable bonds that can be incorporated into polymer
carriers. In the case of polymer–drug conjugates, pH-sensitive Enzymes perform a vast array of important functions
linkages such as hydrazone, hydrazide, and acetal have been inside our body. For example, they can covalently link
used to directly attach drug molecules to polymers. To this polymer chains together to generate self-assembled struc-
Ü
Ü

Angewandte
tures. They can also break certain bonds, causing disassembly infection). Thermoresponsive release relies on a sharp change
or destruction of the structures.[104] The latter enzymatic to the physical properties of a temperature-sensitive material.
reactions can be utilized to trigger the release of a drug. More Such a sharp response can trigger the release of drug in the
significantly, the altered expression of a specific enzyme event of variation to the local temperature around the carrier.
associated with a pathological condition can be employed to The range of temperatures, within which the drug-delivery
achieve enzyme-mediated drug release at the desired biolog- system is activated, should be kept between 37 and 42 8C,
ical target only.[69e] because temperatures beyond this range will cause protein
An enzyme-sensitive release system includes either denaturation or function disruption.[69d, 115]
a structure scaffold that is susceptible to the degradation by The thermoresponsive drug-delivery systems are usually
a specific enzyme or a linker between the drug and the carrier based on liposomes or nanoparticles composed of thermo-
as the product of an enzymatic reaction.[69d] Hydrolases are sensitive polymers. For liposomes, thermoresponsiveness
the most widely used enzymes for such an application, which usually arises from the conformational/structural changes
can break covalent bonds or modify certain chemical groups associated with the constituent lipids to induce variations to
by altering the balance between electrostatic, hydrophobic, the permeability of the lipid bilayers.[116] Thermosensitive
and van der Waals forces, p–p interactions, or hydrogen liposomes (TSLs) represent an advanced system for drug-
bonding.[104b, 105] For example, proteases can induce the release delivery applications related to cancer therapy. Doxorubicin-
of a drug linked to a carrier through a peptide bond; loaded TSLs (ThermoDox, Celsion Corporation) are under
glycosidases can trigger the release from a polysaccharide- investigation in phase II trial for the treatment of breast
based carrier; lipases can facilitate the drug release by cancer and colorectal liver metastasis, and have reached
hydrolyzing the phospholipid building blocks in a liposome; phase III trial for the treatment of hepatocellular carcino-
and hydrolases can be used to maneuver the assembly and ma.[69e] More recently, advanced liposomal formulations have
disassembly of inorganic nanoparticles, as well as the been demonstrated to release their payloads at the onset of
degradation of a gatekeeping material that blocks the pores hyperthermia (ca. 40–45 8C).[117] The thermoresponsive,
of a carrier.[104b, 106] In addition, kinases and phosphatases have bubble-generating liposomal system is a promising exam-
been used to reversibly break/form covalent bonds, achieving ple.[117c–e] This system uses the generation of CO2 bubbles
the release of a drug in an “on–off” manner.[107] through quick decomposition of ammonium bicarbonate
Enzyme-responsive drug-delivery systems have been (NH4HCO3) upon heating to 40 8C. The generated CO2
designed and fabricated in the form of vesicles (micelles bubbles produce a disruptive force to percolate the lipid-
and liposomes), hydrogel nanoparticles, and porous silica bilayer membranes and trigger release of the encapsulated
nanoparticles with an enzyme-sensitive polymer coating as payloads such as proteolytic enzymes[117e] and doxorubi-
the gatekeeper.[105a, 106, 108] All of them have shown promise for cin.[117c,d] On the other hand, thermosensitive polymers
attaining specific release at the inflammation site and inside experience coil-to-globule transition at either a lower critical
a tumor cell. In recent studies, short peptides with sequences solution temperature (LCST) or an upper critical solution
cleavable by matrix metalloproteinases (MMPs) have been temperature (UCST) as a result of change to the efficiency of
used as linkers between poly(ethylene glycol) (PEG) chains hydrogen bonding between the polymer chains and water
and liposomes,[109] polymeric nanoparticles,[110] or iron oxide molecules.[118] The LCST and UCST refer to critical temper-
nanoparticles.[111] Cleavage of the PEG shell in the tumor atures below and above which the polymer is completely
microenvironment led to the exposure of a surface bioactive miscible with the solvent, respectively. When the temperature
ligand, which enhanced intracellular penetration of the is above the LCST, the polymer will become hydrophobic and
nanosized carrier. This approach allowed for the systemic change its conformation from the expanded (soluble) to the
administration of siRNA-loaded nanoparticles at a 70 % globular (insoluble) state.[118] If the polymer is used as
gene-silencing efficiency in tumor-bearing mice.[112] Mesopo- a chemically cross-linked network, a thermally reversible
rous silica nanoparticles (MSNs) grafted with polysaccharide swelling/shrinking of the network will lead to on-demand
derivatives have been demonstrated for the specific delivery release of the encapsulated drug as a consequence of
of doxorubicin through lysosome-mediated cleavage of the controlled changes to the porosity.[119] Thermosensitive poly-
glycoside bonds and reduction of the polysaccharide chain mers can be integrated with other nanosized carriers such as
lengths.[69e, 113] Gu and co-workers recently fabricated nano- liposomes and inorganic nanoparticles to introduce or
particles from peptide dendrimers, which were conjugated enhance the thermoresponsiveness (Figure 5). Examples of
with doxorubicin through an enzyme-responsive tetra-pep- the commonly used thermosensitive polymers include:
tide linker, Gly-Phe-Leu-Gly (GFLG). The nanoparticles poly(N-isopropylacrylamide) (PNIPAAm), poly(N,N-
showed better in vivo antitumor efficacy over free doxorubi- diethylacrylamide) (PDEAAm), poly(methyl vinylether)
cin at equal dose.[114] (PMVE), poly(N-vinylcaprolactam) (PVCL), and poly(ethy-
lene oxide)-poly(propylene oxide)-poly(ethylene oxide)
3.2.3. Thermoresponsive Release (PEO-PPO-PEO, also known as Pluronics).[104b, 120]
Thermoresponsive release is among the most extensively 3.2.4. Photosensitive Release

investigated strategies for smart drug delivery because it can
take advantage of the local temperature increase caused by The use of light as a stimulus to trigger drug release has
the pathological condition (e.g., tumor, inflammation, or been actively explored owing to its convenience for remote
Ü
Ü

.
Reviews
compound,[124] by generating an azobenzene-based molecular

valve at the entrance of each pore,[125] and by inducing light-
controlled host–guest recognition between the cavity in
cyclodextrin and an azobenzene derivative.[126] The hydro-
phobic–hydrophilic transition as a result of trans–cis switching
has also been used for this purpose.[127] Besides these systems
involving conformational/structural changes, UV light has
also been employed to trigger the release of biological
effectors by breaking chemical bonds in a system widely
known as caged compounds.[128] Despite many reports on this
and related systems, the shallow penetration depth
(< 200 mm) caused by the strong scattering of soft tissues
has limited the use of UV-triggered systems for in vivo
applications.[69f] Even for in vitro applications, the UV light
Figure 5. Drug release from two different types of thermosensitive may also cause damages to the cells.
carriers: a) a liposome containing a thermosensitive polymer and b) a Near-infrared (NIR, 700–900 nm) irradiation can pene-
nanoparticle coated with a thermosensitive block copolymer. Upon trate more deeply into soft tissues than UV and visible light
heating, the thermoresponsive component undergoes conformational
without significantly damaging/heating the area of applica-
change, initiating or accelerating the drug release. Panel (a) is
reproduced with permission from Ref. [116], copyright 2013 Elsevier.
tion. The capability of plasmonic nanoparticles to transduce
Panel (b) is reproduced with permission from Ref. [121d], copyright the absorbed NIR light into heat has been used to trigger the
2013 Royal Society of Chemistry. release of drug molecules.[129] For example, doxorubicin-
loaded Au nanocages could be used for controlled release by
irradiation at 808 nm, improving anticancer activity and
potentially reducing systemic toxicity.[129a] A brief discussion
on this system can be found in Section 6.8. The photothermal
effect of Au nanorods upon exposure to NIR irradiation can
also cause rapid rise in local temperature, which has been
exploited to induce dehybridization of DNA helices con-
jugated to the surface and thereby release doxorubicin
molecules bound to consecutive cytosine–guanine base
pairs.[129b] However, inorganic nanoparticles are usually not
biodegradable and raise concerns in their potential long-term
toxicity.[130] Recently, organic transducers that can convert
absorbed NIR light into heat have attracted significant
attention.[131] Among them, indocyanine green (ICG), an
FDA-approved fluorescent dye, is the most widely used
molecule for cancer therapy.[132] For example, PLGA-based
nanoparticles loaded with doxorubicin and ICG showed
strong temperature responsiveness and faster doxorubicin
release under NIR irradiation.[133]
Figure 6. Photosensitive liposomes constructed through incorporation
of light-responsive units into the lipid bilayers with an aim to control
the drug release with optical irradiation. The release can be achieved
through a) photoisomerization, b) photocleavage, and c) photopolyme-
4. In vitro and in vivo Delivery
rization. Reproduced with permission from Ref. [129d], copyright 2012
Ivyspring. Nanoparticles, when empowered with either passive or
active targeting capability, can enhance the concentration of
drugs inside a tumor, while reducing systemic toxicity in
control and its potential of high spatiotemporal resolution healthy tissues. Although the use of nanoparticles as a drug-
(Figure 6).[69d,e, 121] Photosensitivity is often introduced to delivery system offers many advantages, a number of issues
nanoparticles through functional groups that can change remain to be addressed, including their instability during
their conformations or other properties upon irradiation by blood circulation, low renal clearance, limited accumulation
light with a proper wavelength.[122] Notable examples include in cancer tissues, and inadequate uptake by cancerous cells.
azobenzene, pyrene, nitrobenzene, cinnamoyl, and spiroben- To find solutions to these problems, it is of great importance
zopyran.[104b, 123] The reversible molecular switching of the to have a comprehensive understanding of the responses to
azobenzene group (and its derivatives), from trans to cis when nanoparticles by biological systems at the levels of cell, tissue,
irradiated by UV light (300–380 nm) and from cis to trans by organ, and body.
visible light, allows for photoregulated control of drug
release.[69e] This has been realized by functionalizing the
interior pores of MSNs with an azobenzene-containing
Ü
Ü

Angewandte
4.1. In vitro Delivery hydrophobic or electrostatic interactions. Depending on the

size and surface properties of the cargos to be internalized,
Understanding the in vitro delivery of nanoparticles is four major pathways have been identified:[134] 1) clathrin-
a critical initial step toward their successful applications. On mediated endocytosis (also known as receptor-mediated
one hand, the nanoparticles to serve as a drug carrier must be endocytosis, or RME), which involves the inward budding
first evaluated in vitro at the cellular level before they are from the plasma membrane of vesicles (ca. 100 nm in size)
further tested in vivo at the tissue, organ, and body levels. On containing receptors specific to the cargos being internalized;
the other hand, only with sufficient knowledge of nano- such vesicles possess a crystalline coating made of a protein
particle–cell interactions can one start to engineer the complex associated with the cytosolic protein clathrin that is
properties of nanoparticles for optimal delivery in vivo and recruited to assist the internalization process; 2) caveolae-
effective cancer therapy. mediated endocytosis, where extracellular molecules are
Whenever a nanoparticle encounters a cell, it will be internalized upon binding to specific receptors in caveolae,
quickly internalized into the cell through endocytosis. After- the flask-shaped pits (ca. 50 nm in size) in the plasma
wards, it will be transferred to various organelles, including membrane that exist on the surface of many, but not all
endosomes, lysosomes, Golgi apparatus, mitochondria, endo- types of cells; 3) pinocytosis (also known as cell drinking),
plasmic reticulum (ER), and nucleus. During the intracellular a process that begins with the formation of a pocket through
transport process, the nanoparticle has to be degraded or the invagination of the cell membrane in a highly ruffled
disassembled to allow for quick release of its payload. Here region; the pocket is subsequently pinched off into the cell to
we only focus on the major steps involved in the in vitro generate a vesicle (0.5–5 mm in size) that is nonspecifically
delivery of a nanoparticle. At the end of this section, we also filled with a large volume of extracellular fluid, together with
briefly elaborate on the issue of multidrug resistance. substances and small particles present in the fluid; and
4) phagocytosis, a process by which cells actively bind to and
4.1.1. Endocytosis internalize particles larger than about 250 nm in diameter,
such as small dust particles, cell debris, microorganisms, and
In order to deliver the drug to a subcellular target and even apoptotic cells. The last two processes involve the uptake
execute therapeutic functions, the nanoparticle carrier must of much larger areas of cell membrane than the clathrin- and
first cross the plasma membrane of a cell through endocytosis, caveolae-mediated pathways. While pinocytosis can occur in
an energy-dependent process defined as the internalization of all types of cells, phagocytosis can only be performed by
cargo into a cell by engulfment. The endocytosis process can a small set of specialized mammalian cells, such as macro-
be initiated by either highly selective binding between the phages, monocytes, and neutrophils.[135]
ligand attached to the nanoparticle and the receptor present Figure 7 shows a summary of the endocytosis pathways
on the cell membrane, or nonselective binding based on that have been reported for a number of nanoparticles. For
Figure 7. Pathways for the cellular internalization of different types of nanoparticles. The pathway is mainly determined by the size and surface
properties of the nanoparticles, as well as the type (e.g., macrophages vs. endothelial cells) and activation status of the cells. Despite the
significant progress in recent years, the details of uptake routes for some nanoparticles remain elusive. CNT, carbon nanotube; MSN, mesoporous
silica nanoparticle; SPION, superparamagnetic iron oxide nanoparticle. Modified reproduction with permission from Ref. [134a], copyright 2011
Elsevier.
Ü
Ü

.
Reviews
a specific cell type, the internalization pathway of nano- dependent endocytosis pathway, whose mechanism is yet to
particles is largely determined by their size and surface be elucidated.[141a]
properties (hydrophobicity vs. hydrophilicity, the sign/density The endocytosis of nanoparticles is also affected by the
of charge, and the type/density of a ligand). While large type and physiological condition of the cells. For example,
particles are internalized through phagocytosis and pinocy- Chan and co-workers compared the uptake of transferrin-
tosis, small particles have to rely on clathrin- and caveolae- coated Au nanoparticles by three different cell lines: STO
mediated (or occasionally, independent of clathrin and mouse embryonic cancer fibroblasts, HeLa human cervical
caveolae) pathways.[136] The geometry or aspect ratio of cancer cells, and SNB19 human astrocytoma cells. They found
nanoparticles is another factor affecting the mode of cellular that HeLa cells and STO cells exhibited the fastest and
uptake.[137] For example, Lehr and co-workers found that slowest uptake rates, respectively, for the Au nanoparticles,
clathrin-mediated endocytosis is the primary uptake mecha- and the uptake of all Au nanoparticles occurred through
nism for spherical nanoparticles. In comparison, nanoparti- clathrin-mediated endocytosis, regardless of the size of the
cles with a high aspect ratio may have one dimension fall nanoparticles.[142] Finally, the internalization of nanoparticles
within the clathrin limit but the other dimension within the was known to be temperature dependent.[143] When incubat-
limits of pinocytosis and phagocytosis, and therefore the ing PLGA nanoparticles with cells at 4 8C, a condition under
uptake mechanism of these nanoparticles would actually which energy-dependent endocytosis was halted, the uptake
depend on their orientation on the surface of the cells.[137b] of the nanoparticles by the cells decreased dramatically when
Interestingly, simulation further showed that the endocytic compared with the regular cultures performed at 37 8C.
rate of spherical nanoparticles is dependent on their size,
whereas the endocytosis of spherocylindrical nanoparticles 4.1.2. Intracellular Transport
may proceed in a sequence of laying-down-then-standing-up
if the particles are docked on the membrane plane in an initial After internalization, the nanoparticles enveloped by
upright position.[137c] vesicles will be transported along the endolysosomal network
Using superparamagnetic iron oxide nanoparticles to other organelles or sometimes even be exocytosed
(SPIONs) as a model system, Gupta and co-workers demon- (excretion from the cell). As shown in Figure 8, the intra-
strated that the endocytic pathways
of these magnetic nanoparticles
were strongly correlated with their
surface properties.[138] For example,
they investigated the internalization
of SPIONs and pullulan-coated
SPIONs (Pn-SPIONs) by cells
using transmission electron micros-
copy (TEM) and found that fewer
Pn-SPIONs entered the cells as
compared to SPIONs. This reduc-
tion in uptake for Pn-SPIONs can
be attributed to the hydrophilicity
of pullulan, which prevented Pn-
SPIONs from interacting with the
cell membranes.[138, 139] Mirkin and
co-workers reported a scavenger-
receptor-mediated endocytosis
pathway for the cellular uptake of
DNA-coated Au nanoparticles.[140]
They found that serum proteins
hampered the cellular uptake of
DNA-coated Au nanoparticles and
the highest uptake was achieved for
Au nanoparticles under serum-free
cultures. The cellular uptake of
MSNs was also dependent on the
particle size and surface charge.[141]
While MSNs with a low density of
positive charges on the surface were Figure 8. Intracellular transport of nanoparticles. After internalization, the nanoparticle is trafficked
along the endolysosomal network in vesicles with the aid of motor proteins and cytoskeletal
internalized through clathrin-medi-
structures. Note the difference in pH values between different intracellular compartments. ER,
ated endocytosis, particles with endoplastic reticulum; ERC, endocytic recycling compartment; MTOC, microtubule-organizing
a high density of positive charges center; MVB, multivesicular bodies. Modified reproduction with permission from Ref. [143b], copy-
on the surface underwent a charge- right 2011 the Royal Society of Chemistry.
Ü
Ü

Angewandte
cellular trafficking of nanoparticles is a very complex process endolysosome. As an example, QDs with a surface coating
that involves motor proteins shuttling the nanoparticle- of PEG-grafted polyethylenimine (PEI-g-PEG) were capable
loaded vesicles along cytoskeletal structures (e.g., micro- of penetrating cell membranes and then disrupting endolyso-
tubules) within a cell.[143b] somal organelles, owing to the highly positive surface charges
In a recent study, Saltzman and co-workers used rhod- provided by the multiple amine groups.[148] Labhasetwar and
amine-loaded PLGA nanoparticles to explore the intracellu- co-workers fabricated PLGA nanoparticles that were respon-
lar transport processes in three different types of epithelial sive to pH change and studied the internalization of these
cells (HBE bronchial epithelial cells, Caco-2 epithelial color- nanoparticles using vascular smooth muscle cells (VSMCs).
ectal adenocarcinoma cells, and OK kidney epithelial Once internalized, the PLGA nanoparticles underwent rapid
cells).[143a] After internalization, the PLGA nanoparticles endolysosomal escape because of the reversal of surface
could escape endolysosomal degradation, and be sequentially charges on the nanoparticles from anionic to cationic under
transported to the Golgi apparatus and ERs. Based on these the low pH value in endolysosomes and the subsequent
results, an endocytosis–exocytosis pathway was proposed, by induction of the “proton-sponge effect”.[149]
which the PLGA nanoparticles encounter endosomes first Rapid escape/release of nanoparticles from endolyso-
and then escape from the compartment, followed by inter- somes can also be induced when their surfaces are modified
actions with exocytic organelles (i.e., ER, Golgi apparatus, with a pH-sensitive peptide capable of physically interacting
and other secretory vesicles) in the cell. with endolysosomal membranes. GALA, a pH-sensitive
Nie and co-workers used Tat-peptide-conjugated quan- fusion peptide composed of 30 amino acids with repeating
tum dots (Tat-QDs) to examine the intracellular transport of units of glutamic acid-alanine-leucine-alanine, could perturb
nanoparticles in HeLa cells. They took advantage of dynamic the lipid bilayer and facilitate nanoparticles to escape from
confocal imaging and found that Tat-QDs were internalized endosomes at low pH values.[150] When the pH value
through pinocytosis, followed by entrapment in the cytoplas- decreased from 6 to 5 in the endosome, the negative charges
mic organelles. The vesicles loaded with Tat-QDs were found on GALA decreased, causing a conformational change from
to be actively transported along microtubule tracks by random coil to amphipathic a-helix. This change allowed
molecular machines, and finally to the microtubule-organiz- GALA to bind to the endosomal membrane, causing
ing center (MTOC), which is located outside the cell membrane disruption.
nucleus.[144] In many cases, nanoparticles in the endolysosomes or the
Interestingly, the surface of nanoparticles can be func- cytoplasm must be destructed to a certain extent to allow for
tionalized with a ligand to target a specific organelle in the proper release of the payloads. For polymer nanoparticles,
cell. For example, adding a nuclear localization signal (NLS) several strategies can be utilized to improve the efficiency of
peptide motif to the surface of nanoparticles could lead to disassembly, including the use of enzyme-active linkers, acid-
effective nuclear targeting. As reported by Mao and co- labile cross-linkers, pH-sensitive detergent, thermal-sensitive
workers, liposome protamine/DNA complexes termed lip- liposomes, and disulfide cross-linkers that are sensitive to
oplexes (LPDs) were accumulated in the nuclei of cells after a reducing environment.[151] In one example, Ithakissios and
their internalization when the surface of the particle was co-workers studied the use of nanoparticles based on PLGA-
derivatized with NLS peptides.[145] Compared to LPDs with PEG copolymers for the delivery of cisplatin.[152] They found
no nucleus-targeting ligand on the surface, the gene expres- that the intracellular degradation of PLGA-PEG nanoparti-
sion level was significantly elevated when DNA was delivered cles was dependent on their composition. With higher PEG
into the nuclei of the cells. content, the degradation rate of nanoparticles increased,
resulting in a faster release of the encapsulated cisplatin. In
4.1.3. Intracellular Escape and Degradation of Nanoparticles another example, Tasciotti and co-workers reported a drug-
delivery system based on porous silicon nanoparticles, the so-
For successful delivery of therapeutic agents, the nano- called multistage nanovectors (MSVs).[153] According to their
particles also need to be designed with an ability to escape results, the decomposition of MSVs was largely determined
from the endolysosomal network and enter the cytosol, which by the pore size, and MSVs with larger pore sizes were
is the typical working site for most drugs (Figure 8). To degraded more rapidly. Along with the tunable drug-loading
achieve this goal, several strategies have been explored. For capacity of MSVs, the controlled release of a drug could be
example, a type of virus-like nanoparticles was reported, realized through engineering of the pore size of MSVs.
which were capable of fusing with endosomal membranes and
transporting drugs from endosomes to the cytoplasm.[146] 4.1.4. Multidrug Resistance
Alternatively, nanoparticles were coated with a polymer
(typically with amine groups) that has a buffering capacity Multidrug resistance (MDR) is a major problem encoun-
between pH 5.2–7.0 to enable endosomal escape through the tered in chemotherapy that negatively impacts the treatment
“proton-sponge effect”.[147] Once these cationic nanoparticles efficacy of chemotherapeutics.[154] A number of mechanisms
were engulfed into an acidic endolysosomal compartment, the have been reported for MDR, including increased efflux
amino groups could continuously sequester protons pumped pumping of drugs by the overexpressed ATP-binding cassette
inwards by the v-ATPase (i.e., the proton pump), leading to (ABC) transporters, reduced intracellular accumulation of
accumulation of water molecules inside the compartment.[147] drugs by non-ABC drug transporters, blocked apoptosis,
Eventually the swelling resulted in the rupture of the repair of drug-induced DNA damage, metabolic modifica-
Ü
Ü

.
Reviews
MDR.[157] In their study, DOX was used as a chemo-

therapeutic drug and cetyltrimethylammonium
bromide (CTAB), a surfactant commonly used in
the synthesis of Au nanorods, as an MDR inhibitor.
The MSNs co-loaded with DOX and CTAB showed
high therapeutic efficacy against both drug-resist-
ant MCF-7/ADR cells and drug-sensitive MCF-7
cells. The MDR-evading mechanism was shown to
involve a synergistic effect between the cell cycle
arrest and the apoptosis-inducing effect from the
chemosensitization of CTAB.
Liposomes represent another class of delivery
vehicles commonly used for overcoming MDR in
cancer therapy. Galactosylated pluronic P123 (Gal-
P123) modified liposomes loaded with mitoxan-
trone have been used to treat hepatocellular
carcinoma (HCC). An improved therapeutic effi-
cacy against HCC cells was reported, which can be
attributed to the ability of the liposome to reverse
BCRP-mediated MDR.[158] Propylene glycol lip-
osomes loaded with epirubicin (EPI-PG-lipo-
somes) were also demonstrated with a potential
to overcome MDR in breast cancers.[159] Never-
theless, it is interesting to mention that EPI-PG-
Figure 9. Mechanisms of multiple-drug resistance in cancer cells: drug efflux
liposomes were not observed to interfere with P-gp,
caused by multidrug resistance protein (MRP) (e.g., P-glycoprotein or P-gp), down- indicating that this type of nanoparticles might be
regulation of the sensitivity to drug by the tumor suppressor protein p53, reduction able to overcome MDR using a mechanism that
in sensitivity to methotrexate and fluorouracil by phosphorylation of the retinoblas- does not involve the inhibition of P-gp.
toma protein (Rb), and production of resistance to camptothecins through down-
regulation expression or mutations in topoisomerases (Topo).
4.2. In vivo Delivery
tion, and detoxification by drug-metabolizing enzymes While drug delivery in vitro is mainly con-
(Figure 9).[155] Among these mechanisms, the overexpression cerned about nanoparticle–cell interactions, the application
of plasma membrane P-glycoprotein (P-gp, or ABCB1), in vivo emphasizes more on how to send the nanoparticle
a member of the ABC superfamily, is one of the most carrier to the target lesion from the site of administration.
common causes of MDR. P-gp is capable of extruding Upon introduction into the body, the carrier needs to reach
a number of positively charged xenobiotics out of the cell, the target lesion and be accumulated there before any
including some of the commonly used anticancer drugs.[154] treatment can take place. As a result, one has to deal with
The overexpression of other ABC transporters, such as MDR many additional issues related to the transport of nano-
proteins and breast cancer resistance protein (BCRP), has particles, as well as immune response, selectivity and effi-
also been identified as a primary cause of MDR. ciency in targeting, biodistribution, biodegradation, clear-
To suppress MDR of cancerous cells and maximize the ance, and toxicity at the organ and system levels. Ideally, the
cytotoxic efficacy of anticancer drugs, a general strategy is to nanoparticles that serve as the carrier of a drug-delivery
co-administrate one drug (e.g., a gene) to inhibit ABC system should have the following attributes: 1) a good
transporters and promote apoptosis together with another targeting efficiency to ensure selective deposition of drug in
anticancer drug for the actual treatment. To this end, nano- the target lesion while maintaining low concentrations in
particle carriers based on liposomes and polymers have been healthy tissues/organs; 2) consisting of biocompatible and/or
utilized to encapsulate the dual components and at the same biodegradable materials only; and 3) clearance from the body
time ensure precise delivery to the targeted sites.[154] In one within a predetermined time frame.[160] In reality, however, it
example, Amiji and co-workers demonstrated the use of is almost impossible to satisfy all these requirements.
PEO-modified poly(e-caprolactone) (PCL) nanoparticles as With the advances in nanotechnology, many new materi-
a multi-drug-delivery system for the apoptosis modulator als and techniques have emerged to help us realize the
ceramide and the chemotherapeutic drug paclitaxel.[156] Their aforementioned goals. For example, the physicochemical
results indicate that the dual-drug-delivery system could properties of nanoparticles, including composition, size,
greatly improve chemosensitivity of ovarian cancer cells shape, morphology, surface charge, and surface coating, can
exhibiting MDR by bypassing P-gp drug efflux. Alternatively, all be tailored to improve their performance in vivo.[161] An
Shi and co-workers reported a pH-responsive multi-drug- interesting example can be found in the use of QDs for cancer
delivery system based on MSNs that could overcome diagnostics and therapeutics . The long-term toxicity caused
Ü
Ü

Angewandte
by the elements (e.g., cadmium, selenium, and tellurium) blood half-life of neutral nanoparticles is the longest. Altering
commonly used for the synthesis of QDs is difficult to avoid the surface chemistry of a nanoparticle will cause changes to
because these elements are essential to the optical properties its hydrodynamic size and surface charge, as well as reactivity
of most QDs.[162] However, through surface modifications (e.g., binding affinity). Although complete clearance is
such as PEGylation and conjugation with targeting agents, it eventually desired when the treatment is completed, the
is feasible to reduce their accumulation in and toxicity to nanoparticles must be able to avoid rapid clearance in order
major organs to an acceptable level by increasing their to achieve the desired targeting efficiency. In general, the
circulation half-life and reducing their accumulation in organs circulation half-life of nanoparticles should be prolonged to
in a less nonspecific manner.[163] allow them to pass by a lesion multiple times, giving them
Intravenous injection represents the most commonly used increased opportunities to accumulate in the lesion.[170]
route for the administration of nanoparticle-based therapeu- To understand the opportunities and challenges in drug
tics as it bypasses the barriers in the epithelial absorption delivery in vivo, we discuss this subject from two different
process by directly entering the circulatory system.[164] Upon angles: how do nanoparticles reach the targeted lesion and
injection, the nanoparticles are immediately subjected to how are they cleared from the body. After analyzing several
clearance through a joint force of the MPS,[165] the renal biological barriers for the in vivo delivery of nanoparticles, we
system,[166] and the immune system.[167] During circulation, the focus on how to optimize their pharmacokinetics and
size, shape, and surface properties of the nanoparticles can all biodistribution.
strongly affect their behaviors/performance with respect to
targeting and clearance.[136, 168] Figure 10 shows how the 4.2.1. The EPR Effect and Passive Tumor Targeting
behavior and fate of nanoparticles in the body are dependent
on their size, surface charge, and hydrophobicity. In general, Nanoparticle therapeutics directly administrated into the
nanoparticles with a size smaller than 6 nm will be rapidly circulatory system[164] need to extravasate through the vascu-
filtered out and cleared by the kidneys. Nanoparticles larger lar walls into the target lesion and then release the payload.
than 8 nm cannot undergo glomerular filtration; instead, they Unlike small molecules, nanoparticles cannot go through the
will either accumulate in a lesion or be cleared by the tight junctions between endothelial cells on normal vascular
MPS.[165, 169] A positive surface charge on the nanoparticles linings, owing to their relatively large sizes (Figure 11, left
will lead to high systemic toxicity because of complications panel). However, the vessels inside a tumor region are well-
such as hemolysis and platelet aggregation, and the nano- known for their leaky walls (Figure 11, right panel), allowing
particles tend to be quickly cleared from the blood by the nanoparticles with the right sizes to pass through effi-
MPS.[169] Negatively charged nanoparticles have longer circu- ciently.[171] As the lymphatic system inside a tumor is largely
lation half-life than their positive counterparts while the absent or dysfunctional, the insufficient drainage facilitate
Figure 10. The surface charge (zeta potential), size, and surface hydrophobicity of nanoparticle can affect their cytotoxicity (surface reactivity),
recognition by the mononuclear phagocyte system (MPS), clearance (renal or biliary), and enhanced permeability and retention (EPR) effect in
tumor targeting. Modified reproduction with permission from Ref. [201], copyright 2012 American Association for Cancer Research.
Ü
Ü

.
Reviews
Figure 11. Transport of nanoparticles with different sizes and small molecules through normal (left) and cancerous (right) tissues. The enhanced
permeability and retention (EPR) effect is a unique feature of most tumors, allowing nanoparticles of appropriate sizes to accumulate more in
cancerous tissues than in normal tissues.
accumulation of nanoparticles in the tumor tissue.[171, 172] This tumor type, the openings in the tumor vasculature are
phenomenon has been widely known as the EPR effect of typically in the size range of 100–800 nm.[177] A functional
nanoparticles, which is the basis for passive tumor targeting. pore size of 1200–2000 nm has also been reported in MCa-IV
The EPR effect was first noticed by Maeda and co- mouse mammary carcinoma, which is likely at the high end of
workers in studying the inflammation induced by microbial size range for tumor vessel leakage.[175]
infections.[173] In 1986, Matsumura and Maeda further pro- Among various parameters, the size of a nanoparticle
vided experimental evidences to support the concept of the plays the most important role in EPR-based tumor targeting.
EPR effect in tumor targeting for the first time.[37a] Because of As shown in Figure 11, only nanoparticles with a size smaller
the EPR effect, both macromolecular drugs and nanoparticle than the gap between adjacent endothelial cells can extrav-
therapeutics can target tumors more efficiently than small- asate from the vasculature. As described above, the cutoff size
molecule drugs, as the extravasation of nanosized objects typically varies from 100 to 800 nm, depending on the type
occurs in a tumor-selective manner. Over the past several and stage of a tumor. Particles smaller than this cutoff size can
decades, utilization of the high permeability of tumor tissues extravasate from the blood vessels into the tumor intersti-
for nanoparticle delivery has become an important strategy tium. When liposomes of different mean sizes were tested, it
for the design and development of new therapeutics for was shown that the cutoff size for extravasation into tumors
cancer treatment. was approximately 400 nm,[178] whereas particles with diam-
The EPR effect is one of the most important features and eters smaller than 200 nm were found to work most effec-
results of tumor angiogenesis. Solid tumors rely on rapid tively.[5] This is because particles larger than 200 nm were
angiogenesis to maintain sufficient supplies of nutrients and cleared from the blood stream more rapidly by the spleen (see
oxygen.[174] The rapid proliferation of endothelial cells during Section 4.2.3). Once extravasated, the penetration of the
angiogenesis usually results in a reduced density of endothe- nanoparticles into the tumor tissues is a diffusion-mediated
lial cells and thus loss of tight junctions and formation of large process, which is inversely correlated with the particle size.
gaps between the cells. The presence of large gaps between On the other hand, the nanoparticles extravasated into the
the endothelial cells on the tumor vascular walls has been tumor interstitium can also travel back into the blood vessels
confirmed by direct visualization through optical and electron through the gaps in vascular walls and then get cleared by the
microscopy.[175] The underlying basement membrane of the MPS or kidneys.[5] For nanoparticles relatively small in size,
blood vessels is mostly abnormal or missing.[176] In addition, an equilibrium in particle distribution across the vasculature
tumor blood vessels lack pericytes and smooth muscle cell can be rapidly established. In general, nanoparticles with sizes
layers, making them more vulnerable to the high interstitial in the range of 30–200 nm show better retention by the tissue
pressure and rapidly shifting blood flow.[177] Depending on the resistance, shifting the equilibrium toward extravasation and
Ü
Ü

Angewandte
Figure 12. Biodistribution and clearance of nanoparticles from the human body. Tissue defects as well as the size, targeting ligand, and stealth
properties of the nanoparticles are some of the major factors that affect the biodistribution and clearance of nanoparticles.
leading to enhanced accumulation. Taken together, nano- half-life and “stealth” capability, they generally have less
particles with sizes between 30–200 nm are believed to be protein adsorption and thus improved tumor accumulation as
optimal for passive targeting of most types of solid tumors by a result of significant reduction in clearance by the MPS.[170]
capitalizing on the EPR effect (Figure 12).[179] To achieve a maximum therapeutic effect in treating solid
The tumor interstitium also plays an important role in tumors, nanoparticles need to uniformly penetrate deeply
determining the passive targeting efficiency. The tumor into the tumors and then release their payloads. Cellular
interstitium is composed of an elastic network of collagen uptake of nanoparticles is also needed to increase the drug-
fibers filled with hydrophilic fluid.[180] Unlike the normal delivery efficiency while reducing nonspecific accumulation
tissue, there exists a high interstitial pressure in the tumor and the associated issues such as multidrug resistance
interstitium, especially in the central portion of the tumor, (discussed in Section 4.1). Despite its widespread use in the
which tends to work against the extravasation of nano- clinic, the passive targeting strategy has many limitations as
particles. In general, the transport of nanoparticles into the the vessels formed through angiogenesis are not evenly
interstitium is driven by a net force between the extravasation distributed in a solid tumor and the permeability may not be
and interstitial pressure, as well as the gradient in concen- homogeneous throughout the tumor. For a small tumor or
tration.[177, 181] Interestingly, the shape of the nanoparticles was metastatic lesion that does not exhibit strong angiogenesis,
also found to play a role in EPR-based tumor targeting.[182] the passive targeting efficiency based on the EPR effect will
Both simulation and experimental results have shown that be rather limited.[5] Active targeting will help address some of
nanoparticles with a spherical shape tend to follow a laminar these issues.
flow pattern so that only those particles that move near the
surface of the vascular wall will be able to extravasate into the 4.2.2. Active Tumor Targeting
tumor.[7c, 183] In contrast, rod- and bar-shaped nanoparticles
are hydrodynamically more unstable and sometimes fail to To better utilize the biochemical properties of cells to be
follow the flow pattern as they travel in the blood stream.[184] targeted, ligands such as small molecules, peptides, antibodies
These hydrodynamic features provide more opportunities to and antibody fragments, and nucleic acids (e.g., aptamers)
finely tune the geometrical parameters of nanoparticles and have been added to the surface of nanoparticles to improve
thereby enhance their chance to cross the gaps on the vascular their targeting efficiency. This new targeting mode that
wall.[184] involves molecular recognition is known as active targeting,
The surface properties (including functional groups and in which ligand–receptor binding allows the nanoparticle to
charges) may affect the efficiency of extravasation and selectively and strongly bind to the surface of a specific type
retention as they affect the hydrodynamic radius, plasma of cells. This strategy has proven to be effective in vitro and to
reactivity, circulation half-life, and “stealth” capability of the a certain extent in vivo.[5 185] For example, when conjugated
nanoparticles. For nanoparticles with prolonged circulation with a targeting ligand, the nanoparticles often show
Ü
Ü

.
Reviews
enhancement in internalization as this process is dominated effect depends on the properties of both the tumor (e.g., the
by receptor-mediated endocytosis.[186] The conjugated ligand type and degree of angiogenesis and degree of tumor
will increase the affinity of binding and thereby induce vascularization) and the nanoparticles (e.g., the size, shape,
receptor-mediated endocytosis more effectively. For targeting charge, and surface chemistry).[160a] The toxicity profiles and
solid tumors in vivo, the nanoparticles with an active targeting therapeutic effects of most nanoscale therapeutics still need
ligand on the surface still need to rely on the EPR effect to to be improved to meet the minimum requirements for
pass through the gaps in vascular walls. clinical applications.
Currently, the improvement in tumor accumulation ach-
ieved through the introduction of an active targeting ligand is 4.2.3. Clearance by the MPS
still under debate.[187] It is believed that the accumulation of
nanoparticles in tumor tissue is dominated by the passive The clearance of a foreign substance or object from the
process, which is time dependent and requires a long circu- body may involve organs such as liver, spleen, and kidneys, as
lation half-life. As discussed in the previous section, the well as the immune and complement systems (Figure 12). It is
efficiency of this accumulation process is largely determined a natural process that helps maintain our body in a healthy
by the physicochemical properties of nanoparticles other than state. The original targets of these systems for clearance are
the active targeting ligand. Even without using any targeting mostly pathogens such as bacteria and viruses, but they also
ligand, it is always possible to increase the accumulation of have the capability to handle other exogenous particles. The
nanoparticles in a tumor by engineering their size, shape, and force for clearance is so strong that most nanoparticles
surface chemistry.[188] In the presence of a targeting ligand, the administrated intravenously will be removed from the blood
retention and uptake of nanoparticles by cancer cells can in as little as a few minutes to hours. Upon injection, the
indeed be augmented as a result of receptor-mediated clearing process will immediately “compete” with the target-
endocytosis, but only after the nanoparticles have extrava- ing process (either passive or active) for the nanoparticles,
sated from the vasculature.[189] In this way, active targeting can resulting in an unfavorable distribution between the tumor
help achieve a higher intracellular drug concentration, even site and organs such as the liver. The rapid clearance can
though there is only a modest improvement in tumor dramatically offset the desired targeting effect and present
accumulation.[187] The escalation in intracellular drug concen- a major barrier for the development of effective nanomedi-
tration can drastically increase cellular cytotoxicity and cine.[160a]
improve the therapeutic efficacy of drugs that work with The MPS and renal clearances represent two major routes
intracellular targets.[189, 190] Some experimental results suggest for the removal of nanoparticles from our body. One of the
that active targeting would increase both tumor accumulation physiological functions of the MPS is to actively capture and
and cell uptake,[191] but these results are likely caused by the eliminate viruses and other relatively small objects.[193] MPS
discrepancy in surface properties of the particles, non- involves organs such as the liver, spleen, and bone marrow
uniformity of tumor models, and the variance of targeting that are rich in phagocytic cells such as macrophages, Kupffer
ligands.[160a] On the other hand, it is argued that active cells, and monocytes. These cells are able to engulf and digest
targeting may anchor nanoparticles to tumor cells next to the nanoparticles. Studies have shown that the MPS is responsible
leaky vessels, decreasing the efficiency of diffusion, depth of for the clearance of most nanoparticles larger than 10 nm,
penetration, and uniformity of distribution.[7h] To overcome regardless of their shape and surface chemistry.[165]
these potential drawbacks, the density of the targeting ligand When nanoparticles enter the plasma, opsonization (i.e.,
presented on the nanoparticles surface needs to be carefully the adsorption of serum proteins) will occur immediately on
tuned to optimize the balance between penetration depth and their surfaces.[5, 160a] Through opsonization, foreign organisms
binding affinity. or particles will be coated with nonspecific proteins known as
Active vascular targeting has been demonstrated as opsonins to generate a corona and make the particles more
a promising alternative for tumor targeting. By targeting visible to the phagocytic cells in the MPS. Opsonins typically
and killing the endothelial cells of the tumor vessels, nano- contain complement proteins and immunoglobins (usually
medicine can be used to eradicate tumor cells by cutting off IgG) along with albumins, fibronectins, fibrinogens, and
their supplies of oxygen and nutrients.[192] In the case of active apolipoproteins.[194] Studies have shown that the corona has
vascular targeting, the targeting ligand is of critical impor- a layered architecture. It starts with an inner layer of proteins
tance as tumor accumulation is no longer determined by the that strongly adsorb onto the surface, with Kd 10 6 to 10 8 m,
EPR effect, but by the binding affinity to blood vessels. to form the hard corona, which is then surrounded by a layer
Nanoparticles have been developed for targeting various of soft corona formed by weak interactions.[169, 195] The
moieties on the vascular walls, including fibronectin extra- primary driving forces for opsonization are based on hydro-
domain B, large tenascin-C isoforms, integrins, annexin A1, as phobic and electrostatic interactions, together with entropic
well as vascular endothelial growth factors (VEGFs) and their and conformational changes for the adsorbed proteins.[196]
receptors.[192] Depending on the charge and hydrophobicity of the nano-
Currently, the targeting of solid tumors remains a chal- particles, opsonization can occur within minutes. Experimen-
lenging task and represents a bottleneck for the development tal results suggest that a charged surface tends to be covered
of future cancer therapeutics. Even those “successful” drug- by proteins more rapidly than their counterparts with a neutral
delivery systems based on nanoparticles still show significant surface.[160a]
accumulation in major organs. In general, the therapeutic
Ü
Ü

Angewandte
Upon opsonization, the nanoparticles will be rapidly targeting efficacy. If possible, the surface of the nanoparticles
internalized by phagocytic cells in MPS organs. As a result, should be kept neutral, as a neutral surface attracts the least
the fate of nanoparticles is determined by their surface amount of serum proteins. The presence of negative charges
properties in conjunction with opsonization.[197] Phagocytic on the surface will induce opsonization and thus MPS uptake,
cells have ciliated borders and stellate branches, which but at a reduced rate relative to a positively charged
provide them with an efficient mechanism for trapping and surface.[201] Surface modifications such as PEGylation are
removing foreign particles.[198] These cells possess numerous widely used to reduce opsonization and consequently increase
membrane receptors, including those for the complement the circulation half-life of nanoparticles. Known as a “stealth”
proteins and for the Fc portion of IgG that is deposited on the property, PEGylated nanoparticles are less recognized by
nanoparticles.[198] After binding of the nanoparticles to the phagocytic cells as a result of lower opsonization and tend to
receptors on phagocytic cells, receptor-mediated endocytosis accumulate in tumors more effectively.
will be initiated, followed by enzymatic degradation. Seg- Recent studies have demonstrated that the opsonization
ments that remain after enzyme breakdown will be retained in process could be much more complicated than simple protein
the cells and be accumulated in organs such as the liver and adsorption. Nanoparticles could be actively targeted by the
spleen (which is typically the situation for noble-metal immune system through antibodies. It has been shown that up
nanoparticles).[198] to 25 % of patients developed anti-PEG antibodies after
The liver plays the most important role in cleaning treatment with PEGylated nanoparticles.[167] This finding is
nanoparticles that do not undergo renal clearance, as it critically important as PEGylation is heavily relied upon to
contains numerous Kupffer cells whose natural function is to provide the nanoparticles with a “stealth” property. Further
eliminate foreign substances through phagocytosis. Hepato- studies indicate that the balance between Th1-Th2 cytokines
cytes can also take up and process nanoparticles, providing and M1-M2 macrophages also affects the rate and amount of
the critical function of biliary excretion, which can expunge nanoparticle clearance, because Th1-prone mice cleared
certain nanoparticles permanently from the body.[198] All nanoparticles at a slower rate than Th2-prone mice.[202] All
nanoparticles excreted through the biliary system must be these results suggest that the immune system may actually
broken down by hepatocytes before secretion, so clearance affect the uptake and clearance of nanoparticles in a more
from the biliary system is considered to be an active process, profound way than what is currently understood.
even though hepatocytes are not a part of the MPS. It is worth
mentioning that the processing of nanoparticles by hepato- 4.2.4. Renal Clearance
cytes (i.e., the hepatic process), and the biliary excretion
process are relatively slow, although hepatic uptake of Renal clearance is based on physical filtration (dialysis)
nanoparticles always occurs quickly.[193] Different from the rather than cell uptake. Unlike the clearance of nanoparticles
hepatic process, nanoparticles processed by Kupffer cells and from circulation by the MPS system, the renal system removes
phagocytic cells will always stay in the MPS organs. In the nanoparticles from the body through the urine rather than
general, there is always a large number of nanoparticles having them accumulated in related organs. As the removal of
accumulated in the liver and other MPS organs, which could nanoparticles by bile is a relatively long process, renal
potentially induce long-term side effects. clearance is an optimal method for expelling nanoparticles
Intrinsically a cleaning organ, the spleen has a blood from the body with minimal side effects.
filtration system composed of a tight reticular mesh (ca. Renal clearance of nanoparticles is a passive process,
200 nm wide) made of interendothelial cells.[199] Nanoparticles involving glomerular filtration and tubular secretion. After
larger than 200 nm are preferentially cleared by spleen. By entering the glomerular capillary bed, nanoparticles are either
choosing an appropriate size and surface chemistry, rigid filtered out through the glomerular capillary or remained
nanoparticles with long circulation half-life can accumulate in within the vasculature, depending on their properties
the spleen at a high percentage.[200] Experiments showed that (Figure 13). During extraction, the nanoparticles need to be
about 50 % of polystyrene nanoparticles with a diameter of filtered through the fenestrated endothelium, the glomerular
250 nm and coated with poloxamine 908 accumulate in the basement membrane (GBM), and the glomerular epithelial
spleen within 24 h after injection because of the physical cells.[203] Although these layers of cells are known to have
filtration effect.[199, 200] Such a feature mainly exists in nano- pores with sizes up to 43 nm, the functional or physiologic
particles with long plasma half-life, as other types of nano- pore size is only around 5 nm if we take into consideration the
particles tend to be captured by the MPS in the liver because combined effect of all layers in the glomerular capillary
of their high phagocytic activity after opsonization. In wall.[203, 204] As a passive process, the filtration of particles is
addition to the active capture by the MPS, clearance of highly dependent on their sizes.[203] Typically, nanoparticles
large nanoparticles (> 150 nm) also utilizes physical filtration with a hydrodynamic diameter of less than 6 nm are rapidly
to trap particles in the spleen or liver,[199] making clearance by filtered out, whereas those with hydrodynamic diameters of
the major MPS organs a hybrid of active and passive 6–8 nm are removed less efficiently, and particles larger than
processes. 8 nm generally cannot escape.
To avoid clearance by the MPS, the surface of nano- The difference in particle properties can often result in
particles needs to be carefully engineered to prevent or at distinct renal handling. Several studies using QDs and Au
least mitigate opsonization. This will help prolong their nanoparticles have been conducted to evaluate the effect of
plasma half-life, reduce MPS clearance and enhance the these characteristics on the size threshold for filtration.[160b, 166]
Ü
Ü

.
Reviews
through, while both positively and negatively

charged nanoparticles adsorbed more serum pro-
teins,[166] increasing their apparent hydrodynamic
diameters and dramatically reducing their ability
to go through renal filtration.[166] PEGylation has
been proven effective in preventing protein
adsorption, but the PEGylation process itself
increases the hydrodynamic diameter, thus nega-
tively impacting its usefulness in renal filtration.[166]
For nanoparticles with a size of around 6–8 nm, the
surface charge and surface chemistry are especially
important, as these particles are no longer small
enough for charge-independent filtration.[203]
Before draining into urine, nanoparticles may
still have a chance to be resorbed from the tubular
fluid. Studies have shown that some nanoparticles
based on polyamine dendrimers may undergo
proximal tubule resorption.[165] Further studies
are still needed to evaluate the resorption of
nanoparticles as the capability of nanoparticles to
be resorbed is still an open question.
It is reasonable to conclude that nanoparticles
with hydrodynamic diameters smaller than 8 nm
Figure 13. Renal handling of nanoparticles with different sizes and charges. a) The
can be cleared through the renal pathway when the
glomerular capillary wall contains a filtration slit, which has a physiologic pore size surface chemistry and charge of the nanoparticle
of 4.5–5 nm. Nanoparticles < 6 nm (red) are small enough to be freely filtered, are optimized. Nanoparticles larger than 8 nm can
irrespectively of surface charge. However, positively charged nanoparticles of 6– also undergo renal clearance if they can be some-
8 nm (purple) are more readily filtered than equally sized but negatively charged how broken down into fragments smaller than
nanoparticles. Nanoparticles > 8 nm cannot undergo glomerular filtration. b) As the 6 nm after drug release.
brush border of the proximal tubule epithelial cells is negatively charged, positively
charged nanoparticles are more readily resorbed than negatively charged nano-
particles with comparable sizes. Modified reproduction with permission from 4.2.5. Pharmacokinetics and Biodistribution
Ref. [165], copyright 2008 Future Medicine.
As many anticancer drugs are highly toxic, it is
desirable to have the drug deployed to the tumor
Clearance studies with QDs showed efficient renal excretion site only and then purged within a certain period of time to
for particles with hydrodynamic diameters less than 6 nm minimize the side effects. For nanoparticle-based therapeu-
(4.36–5.52 nm).[166] QDs with a hydrodynamic diameter larger tics, the pharmacokinetics and biodistribution are the results
than 8 nm (8.65 nm) did not go through renal filtration, but of multiple entangling factors, including the targeting effi-
instead were cleared from the circulation by the MPS.[166] ciency and clearance of the nanosized carriers, as well as the
Other studies indicate that Au nanoparticles with a spherical release profile and related properties of the loaded drug. In
shape and a size of 2 nm were rapidly extracted, with only general, the pharmacokinetics and biodistribution of such
3.7 % accumulation in the liver. In comparison, 4.0 % and a therapeutic agent are largely determined by the nano-
27.1 % of Au nanoparticles of 6 nm in size were found in the particle carrier itself up until the drug molecules are
urine and liver, respectively, whereas 0.5 % and 40.5 % of Au released.[205] This makes it more complicated to manipulate
nanoparticles of 13 nm in size ended up in the urine and liver, the pharmacokinetics and biodistribution of nanomedicine,
respectively.[160b] Polyamidoamine (PAMAM) dendrimers because both the distribution of nanoparticles and the release
with a near-spherical shape were also used to evaluate the profile of the drug need to be taken into consideration.[201] For
threshold, and dendrimers of 5.4 nm in size demonstrated intravenously administrated nanoparticles, their distribution
effective glomerular extraction.[165] These experimental in the body is largely controlled by a limited set of organs and
results indicate that the size threshold for renal filtration is the diseased lesion (Figure 12). To a certain extent, the
comparable to that of most macromolecules. pharmacokinetics and biodistribution of a nanosized thera-
In addition to the size, the surface charge is another factor peutic agent would be determined by the amount of nano-
that can influence renal handling of nanoparticles. As the particles distributed among these sites as a function of time.
surface charge of a particle increases, the adsorption of serum Although guidelines have been established for maximizing or
proteins will likewise increase, resulting in a larger in vivo minimizing the accumulation of nanoparticles at these sites, it
hydrodynamic diameter.[166] Moreover, the surface charges on remains a grand challenge to achieve the optimal conditions
the nanoparticle can interact with the charges on the for the accumulation/clearance at all these sites. At the
glomerular capillary wall.[203] As a result, QDs with a neutral current stage of development, the majority of the adminis-
surface charge were shown to have the highest chance to pass tered nanoparticles end up in healthy organs and tissues, even
Ü
Ü

Angewandte
with the assistance of effective targeting,[201, 206] and the off- anatomical features may react differently to the presence of
target toxicity is still unacceptable for most nanoparticle- the same nanoparticles.[212] Nanoparticles engineered for
based therapeutics.[201] certain applications in specific tissues may not be suited for
Animal models can affect pharmacokinetic studies in other applications involving other types of tissues. 2) The
many ways. For different kinds of laboratory animals, the intrinsic properties associated with the material of a nano-
primary MPS organs of nanoparticle sequestration are species particle are not necessarily a major factor in determining the
dependent.[201] For different animal xenograft models, the biocompatibility of the nanoparticles.[212] 3) When assessing
functional pore sizes of tumor vasculature can also vary the biocompatibility, it is necessary to gauge benefits versus
significantly.[175] As the EPR-based targeting process is highly risks. Hazards such as inflammation may not damage
dependent on the vascular pore size, it is important that the neighboring tissues and will heal over time.[212]
feature of tumor vasculature should resemble the clinical Among biocompatibility issues, toxicity is the most
cases as much as possible.[178] In general, the pharmacokinetics important one and rightfully garners the most attention.
and biodistribution of nanomedicine must be systematically The toxicity of various types of nanoparticles, including those
studied case by case. It has been found that formulations that made of polymers, magnetic materials, noble metals, and
were successful in multiple xenograft models were generally semiconductors, has been summarized and reviewed else-
found to have better therapeutic effects clinically.[205] where.[213] The in vivo toxicity profiles that have been inves-
It is worth noting that several patient-related factors can tigated include skin sensitization, dermal toxicity, ocular
also affect the pharmacokinetics and biodistribution, thus toxicity, inhalation toxicity, oral toxicity, neurotoxicity, devel-
causing pharmacokinetic variability. For example, clinical opmental toxicity, reproduction toxicity, and genotoxicity.[214]
studies show that the age, body composition, gender, and The intrinsic toxicity of materials, including those of the
presence/absence of a tumor in the liver can all alter the nanoparticle and its payload, the corresponding responses
pharmacokinetics of PEGylated liposomal agents.[201] A study from the body, and any reactive species generated may all
involving PEGylated liposomal doxorubicin (Doxil) and play important roles in determining the toxicity in vivo. It is
PEGylated liposomal CKD-602 (S-CKD602) indicates that worth noting that long-term studies (months to years) still
the clearance was dramatically lower in patients over 60 years need to be conducted for most nanoparticles.[160a] It is possible
old,[207] and female patients tended to show a lower clearance that some nanoparticles considered to be non- or minimally
of drugs encapsulated in PEGylated liposomes. Population toxic may induce a higher degree of toxicity during their
pharmacokinetic studies further suggested that patients with degradation or processing by the organism.[160a]
primary or metastatic tumors in their livers, along with It is also worth pointing out that the toxicity can be caused
refractory solid tumors, had a higher clearance for S- by chemical residues left from a synthesis. Complete removal
CKD602.[208] of these residues is often difficult and sometimes impossible.
For instance, CTAB is widely used in the synthesis of Au
4.2.6. Biocompatibility and Biodegradation nanoparticles, in particular, nanorods. Complete removal of
CTAB is difficult and may lead to aggregation of the
Biocompatibility has been a subject of extensive research nanoparticles if completely removed. The positive charge of
ever since the first foreign material was implanted into the CTAB attached to the surface of nanoparticles can induce
human body. As nanoparticles and bulk materials have cytotoxicity and rapid opsonization, followed by MPS clear-
different properties, the responses of the body can be ance.[215] For this reason, new protocols of synthesis involving
drastically different even for those with the same composi- reagents extracted from natural sources as stabilizers and
tion, resulting in different levels of toxicity. For example, Au reducing agents have been developed to produce Au and Ag
has been extensively used in dentistry for centuries, and it is nanoparticles, as well as CdSe QDs.[216] Although the results
widely known to be bioinert and biocompatible.[209] Con- are promising, greater effort is needed for such syntheses to
versely, Au nanoparticles may exhibit toxicity because of their better control the size, shape, and uniformity of the nano-
ability to induce the generation of reactive oxygen species particles.
(ROS).[210] Biodegradability is another important issue in nanomedi-
According to Kohane and Langer, the biocompatibility of cine. In general, it is desirable for the nanoparticles to be
a material can be generally considered as “an expression of completely broken down and expunged from the body after
the benignity of the relation between a material and its the payload has been released. To this end, nanoparticles have
biological environment”.[211] Top issues concerning biocom- been developed from a number of biodegradable polymers
patibility include acute and long-term toxicity, and also the such as PLA, PGA, PLGA, PCL, poly(alkyl cyanoacrylates)
response from the whole body and the functionality of the (PAC), chitosan, and gelatin.[217] A large array of anticancer
tissues/organs involved. In general, biologically inert nano- drugs, including 9-nitrocamptothecin, paclitaxel, cisplatin,
particles that do not induce unacceptable toxic, immunogenic, xanthones, Rose Bengal, triptorelin, and dexamethasone,
thrombogenic, and carcinogenic responses tend to show have all been encapsulated in those biodegradable nano-
a high level of biocompatibility. particles.[59a] To demonstrate biodegradability and acquire the
Several factors must be kept in mind when the biocom- drug release profile while evaluating any therapeutic
patibility issue is evaluated: 1) The biocompatibility of nano- improvement, several types of tests have been designed and
particles is strongly correlated with the type and anatomical conducted both in vivo and in vitro for these systems.[213b] It
features of the surrounding tissue. Tissues with different was shown that these systems could indeed be broken down to
Ü
Ü

.
Reviews
small molecules, which could then be processed by catabolic drawbacks, including the lack of dedicated optical/electric/
mechanisms in the body.[165] Although made of biodegradable magnetic properties, lot-to-lot variability, immunogenicity,
materials, these nanoparticles still exhibited toxicity to inadequate biomechanical properties, and structural com-
a certain extent because of nonspecific accumulation and plexity.[225]
thus release of their toxic payloads in healthy tissues.[218] As a major advantage, natural materials can be readily
Gold nanoparticles have been actively explored for metabolized by and cleared from a biological system through
biomedical applications, including drug delivery, imaging enzymatic or hydrolytic degradation.[225] While this may not
contrast enhancement, and cancer treatment.[219] These nano- be desirable for permanent and long-term implants, such as
particles are normally considered not to be biodegradable as hip replacements, it is a clear advantage when timed
the nanoparticles cannot be readily digested and the resultant biological resorption is desired. Natural materials can be
metal ions can be highly toxic.[220] However, Sokolov and co- chemically modified and cross-linked to adjust degradation
workers recently demonstrated the synthesis of Au nano- rates for specific drug-delivery applications.[226] The most
particles through the assembly of Au clusters, and such frequent concern about natural materials is the immunogenic
nanoparticles are potentially biodegradable in vivo.[221] The response, which can rapidly occur upon introduction into the
Au clusters used in this synthesis had an average diameter of body. This response stems from the fact that the introduced
4 nm, which fulfills the requirement for rapid renal clearance. materials, although similar to endogenous host extracellular
By carefully tuning the ratio of Au clusters to a polymeric matrix components, may not be identical and in fact often
stabilizer, the product with a size of 83 nm showed good contain antigenic contaminants. This issue occurs most
biodegradability in vitro.[221] commonly among protein-derived materials and is typically
In summary, the biocompatibility of nanoparticles less severe for polysaccharides such as chitosan.[225] This
depends on their structure and surface properties, and many immunogenic effect can be reduced through either chemical
other factors. At the current stage of development, long-term modification or purification to remove the immunogenic
assessment (months to years) are missing for most samples. components. However, the complex structures of natural
Therefore, the long-term fate and toxicity of nanoparticles are materials can complicate the modification processes that are
essentially unknown.[222] This is an especially important issue relatively simple to perform with synthetic materials. Despite
for nanoparticles made of non-biodegradable materials.[223] In this, many groups have demonstrated successful procedures
addition, the evaluation of biocompatibility must be carried for the modification (and purification) of natural materials.
out case by case with a systematic methodology and a long- Another common issue arising from naturally derived mate-
term mindset. rials is lot-to-lot variations in molecular structure as a result of
different animal sources. The inconsistency arises not only
from interspecies variations, but also at the tissue level, which
5. Perspectives on the Design of Nanoparticle can complicate processing and quantification of these materi-
Carriers als.[225] Recently, bacterial recombinant techniques have been
used to produce several natural materials, including hyalur-
The first attempted development of a nanoparticle-based onic acid and collagens, effectively addressing the variability
therapeutic can be traced back to the synthesis of a polymer– and immunogenicity issues associated with these materials.
drug conjugate in the 1950s.[224] Ever since, nanoparticle Currently, drug delivery is dominated by nanoparticles
carriers have been prepared and tested using a wide variety of based on synthetic materials because they offer precise
materials, including proteins, polysaccharides, synthetic poly- control over the physicochemical properties of the formula-
mers, metals, and many other organic/inorganic materials. As tions. To deliver anticancer drugs to the tumor sites in vivo,
a major requirement for the design of nanoparticle carriers the nanoparticles must be stable, biologically inert, and
for drug-delivery applications, the composition, size, shape, nontoxic. Concurrently, they must remain in the bloodstream
surface properties, biocompatibility, and degradation profile for a sufficiently long period of time to reach the target site
all need to be precisely engineered and optimized to achieve and even pass by the target site multiple times. The nano-
site-specific release of drugs at therapeutically optimal rates particles can induce the formation of a corona of serum
and dose regimes. proteins around the surface, so highly charged nanoparticles
are phagocytosed by the MPS more quickly than neutral
particles.[195, 223b, 227] Using synthetic materials, the surface
5.1. Natural versus Synthetic Materials charges and hydrophobicity of nanoparticles can be conven-
iently adjusted and optimized to increase their circulation
The use of natural materials is attractive because of their half-life. In addition, their surface functionality can be readily
abundance, good biocompatibility, and the potential to be engineered to maximize their affinity toward the targeted
modified through chemical/biochemical reactions.[175] Natu- receptors.
rally occurring materials offer many advantages over their
synthetic counterparts. For example, the biological system can
easily recognize and metabolically process natural materials 5.2. Size and Shape
through established pathways, while synthetic materials may
induce toxicity, chronic inflammation, and clearance issues. The size and shape, as well as the uniformity, are two
However, natural materials are also plagued by a number of important parameters of a drug-delivery system based on
Ü
Ü

Angewandte
nanoparticles, as they determine the in vivo distribution, particles from block copolymers containing both hydrophilic
toxicity, and targeting ability.[228] Additionally, they can and hydrophobic segments.[238] Studies by Elsabahy and
influence drug loading, drug release, and in vitro and in vivo Wooley suggested that the surface chemistry of nanoparticles
stability. For example, smaller particles have a greater risk of can greatly impact their toxicity, immunogenicity, and biodis-
aggregation during storage and incubation in vitro, but tribution; excess positive charges tend to result in rapid
typically have a longer circulation half-life in vivo. The opsonization and clearance.[239]
degradation of polymer nanoparticles can be strongly affected
by their size as a result of water availability and removal of 5.3.1. Surface Charges
degradation products.
Many studies have demonstrated that nanoparticles have The zeta potential of a nanoparticle is commonly used to
a number of advantages over their micrometer-sized counter- characterize its surface charge.[240] This variable reflects the
parts with sizes in the range of 0.1–100 mm for drug-delivery electrostatic potential of a particle and is influenced by the
applications.[229] Generally, nanoparticles have relatively composition of the particle as well as the medium in which the
higher intracellular uptake and broader availability to nanoparticle is suspended. Nanoparticles with a zeta potential
a range of biological targets owing to their small sizes and above 30 mV (either positive or negative) have been shown to
increased mobility. For example, Amidon and co-workers be stable in suspensions, as repulsion forces originating from
found that nanoparticles of 100 nm in size had a 2.5 times the surface charges can prevent the particles from aggrega-
greater uptake than microparticles of 1 mm in size and 6 times tion. Additionally, the inner surface of blood vessels and the
greater uptake than microparticles of 10 mm in size for Caco-2 surfaces of cells contain various types of negatively charged
human epithelial colorectal adenocarcinoma cells.[230] In species, which repel negatively charged nanoparticles. When
a subsequent study, nanoparticles were found to penetrate the surface charge of nanoparticles becomes higher (either
through the submucosal layers in a rat in situ intestinal loop positive or negative), they will become more easily scavenged
model, whereas micrometer-sized particles were largely by macrophages, resulting in greater clearance by the MPS.
localized in the epithelial lining.[231] It was also reported that Therefore, control over the surface charge can help minimize
nanoparticles can cross the blood–brain barrier (BBB) by the nonspecific interactions between nanoparticles and the
passing the openings of tight junctions treated with a hyper- MPS, preventing the loss of nanoparticles in undesired
osmotic mannitol solution. Nanoparticles coated with Tween locations.[241] Complete exclusion of nonspecific interactions,
80 have also been shown to cross the BBB.[232] As such, however, is currently unattainable.
nanoparticles may provide sustained delivery of therapeutic
agents for difficult-to-treat diseases such as brain tumors.[233] 5.3.2. PEGylation
In fact, some cell lines were found to only take up nano-
particles, while rejecting larger ones.[234] In order to increase the tumor targeting efficiency, it is
The shape of nanoparticles is of equal importance as their necessary to prolong the circulation of nanoparticles in the
size in drug delivery. While spherical nanoparticles are good bloodstream by minimizing opsonization. The most com-
candidates for drug delivery, anisotropic structures can some- monly used approach to achieve this goal is to coat the surface
times provide higher efficiencies because of their larger ratios of nanoparticles with a hydrophilic brush made of PEG
of surface area to volume, as illustrated in Figure 1. The chains.[228]
anisotropy in structure may allow the carrier to take a more Studies have shown that the conformation of PEG on the
favorable configuration for binding with the cell, although the nanoparticles surface is of the utmost importance in repelling
sharp edges and corners can potentially induce injuries to opsonins. While PEG coatings with a brush-like configuration
blood vessels.[235] The mechanisms by which nanoparticles reduce phagocytosis and complement activation, those in
cross the cell membranes has been a subject of extensive a mushroom-like configuration are potent complement acti-
research in recent years, because an understanding and vators to induce phagocytosis.[237] Since the initial use of PEG
control of cellular uptake is important for the development in extending the circulation half-life of a protein,[242] PEGy-
of more effective nanomedicine.[235, 236] For a more detailed lation has been widely adopted to protect nanoparticles such
discussion, please refer to Section 4.1. as liposomes,[243] polymer nanoparticles,[63a] and micelles[244]
from premature clearance during circulation. The PEG chains
form a hydrated shell that allows the nanoparticle to evade
5.3. Surface Properties opsonization and subsequent phagocytosis.[245] However, this
protective shell can interfere with the interactions between
In addition to both size and shape, the surface character- a nanoparticle and the target cell.[246] For example, PEGylated
istics of nanoparticles represent another critical parameter in liposomal doxorubicin showed a prolonged plasma half-life,
determining their drug-loading efficiency and release profile, which is believed to correlate with better therapeutic efficacy.
circulation half-life, tumor targeting, and clearance from the However, the formulation resulted in lower tumor accumu-
body. Ideally, the nanoparticles should have a hydrophilic lation than the same liposomes with no PEG coating,
surface to resist the adsorption of plasma proteins and thus indicating a counterproductive effect of PEGylation.[247] A
escape the uptake by macrophages.[237] This can be achieved in recent publication also noted that PEGylated, multifunc-
two ways: coating the surface of nanoparticles with a hydro- tional envelope-type nanodevices were less effective in
philic polymer such as PEG, or directly fabricating nano- delivering genes to liver cells in vivo than those with no
Ü
Ü

.
Reviews
PEG coating because of the ineffective uptake of the polysaccharides are still unclear. A subtle difference in
PEGylated nanodevice by hepatocytes.[248] Using a computa- molecular weight, the arrangement of monomers, and the
tional model, Bunker and co-workers further demonstrated degree of branching can all result in significant differences in
that some targeting moieties could lose their functionality biological activities. A complete understanding of the mech-
because of steric hindrance from the PEG layer.[249] The anisms of biological effects is the prerequisite for successful
increased stability of nanoparticles by PEGylation can also introduction of polysaccharides into nanomedicine applica-
hinder the endosomal escape, a critical step for effective tions.
intracellular delivery of gene drugs and other therapeutics.[250]
These obstacles have prompted a search for new strategies 5.3.4. Conjugation with Targeting Ligands
and compounds to disguise the nanoparticles. Examples of
recent efforts include the use of different polymers of Many techniques and tools are currently available to
synthetic or natural origin, biomimetic coatings, and the armor nanoparticles for active targeting of cancerous cells.
conditional removal of PEG layers.[251] Traditionally, monoclonal antibodies have been used to target
epitopes on the surface of cells, but the extensive screening of
5.3.3. Polysaccharides peptide and aptamer libraries has greatly expanded the
repertoire of ligands available for targeted delivery.[6] The
As a major class of natural polymers, polysaccharides currently used targeting ligands include antibodies, antibody
have been widely used in drug delivery and tissue engineering fragments, peptides (e.g., RGD for avb3 integrin), aptamers
because of their good biocompatibility, availability, and easy (e.g., those for prostate-specific membrane antigen and
modification.[252] As a result of their capability to avoid the VEGF), oligosaccharides, and even small molecules (folate
complement system and opsonization, some polysaccharides, and SV-119),[185c,d] as long as they can specifically recognize
such as dextran and heparin, have also been recognized as and bind to an overexpressed target on the cell surface. Here
stealth-coating materials.[253] Some studies have shown that we only provide a brief discussion on these ligands:
the polysaccharides, such as chitosan and hyaluronic acid, 1) Monoclonal antibodies (MAbs) are macromolecules
even display certain ligand activities of their own. Nano- widely used as targeting ligands because of their immediate
particles coated with these polysaccharides show more availability and their high affinity and specificity for molec-
efficient cellular uptake than other nanoparticles because of ular targets. These ligands usually possess a molecular weight
specific interactions with various receptors on the surface of of approximately 150 kDa and exhibit high binding affinities.
target cells.[254] Therefore, polysaccharides have gained To date, MAbs have been conjugated to essentially all
increasing interest in the development of nanomedicine as different types of nanoparticles, such as SPIONs,[255]
an effective surface modification strategy. QDs,[256] liposome,[257] and Au nanocages,[258] to give them
Nanoparticles incorporating polysaccharides can be pre- site-specific targeting ability. However, the bulky size and
pared using many different methods, which have been redundant constant region may cause some major issues in the
extensively reviewed.[253a, 255] Polysaccharides can be applied use of MAbs as targeting ligands because of their immuno-
as surface coatings on nanoparticles through electrostatic genicity and size increase (i.e., the overall size of nano-
interactions or directly incorporated into the nanoparticles particles will dramatically increase). The use of antibody
during synthesis. Alternatively, hydrophilic polysaccharides fragments, affibodies, and peptides may help overcome this
can be grafted to hydrophobic molecules, such as cholesterol, shortcoming.
and then used to form nanoparticles through self-assembly, 2) Single-chain variable fragments (scFv) are fusion
which can also encapsulate hydrophobic drugs in the core. proteins of the variable regions of the heavy and light
Furthermore, nanoparticles can be prepared through conju- chains of an antibody (VH and VL) connected with a short
gation of polysaccharides to synthetic polymers. For example, linker peptide of 10–25 amino acids. The molecular weight of
grafting polysaccharides to the side chains of a linear hydro- an scFv is about 27 kDa. By engineering the MAbs to cut
phobic polymer can generate a branched copolymer, and down the redundant parts of the scFv, the size and immuno-
conjugation of the polysaccharide terminus to a linear hydro- genicity of the original antibody can be largely reduced.
phobic polymer can generate a linear diblock copolymer. 3) Affibodies are small, stable Z-domain scaffolds con-
Several hurdles must be surpassed before polysaccharides sisting of 58 amino acids and derived from the IgG binding
can be effectively applied as drug carriers. First, most domain of staphylococcal protein A. The binding pocket is
polysaccharides are of natural origin, and there is a high composed of 13 amino acids and is able to bind to a variety of
degree of variability with respect to the molecular weight and targets, depending on the randomization of the amino acids.
structure depending on the source. These properties critically In contrast with IgGs, the small size (6–15 kDa) of affibodies
determine the biological activities of polysaccharides, and enables penetration into tumor tissue. Affibodies possess
alternative methods need to be established to produce a high receptor affinity, which mimics the active portion of the
polysaccharides with consistent properties. Second, the Fab region of the corresponding antibody. Their short plasma
desired effect of the polysaccharides may be counteracted half-life makes them good candidates as tumor imaging
by the biologically active contaminants of polysaccharide, probes, but less ideal for tumor targeting, where long
such as endotoxins and pathogens. More effective methods circulation half-life is required.[259]
for purifying polysaccharides are urgently needed.[225] Third, 4) Peptides represent a viable targeting moiety with
the exact mechanisms of the biological actions of most several advantageous characteristics, including low molecular
Ü
Ü

Angewandte
weight (ca. 1 kDa), tissue penetration capability, lack of Many recent studies have examined different techniques
immunogenicity, ease of production, and relative flexibility in for the fabrication of polymer nanoparticles, including
chemical conjugation processes.[259] Various peptides that can polyelectrolyte complex formation, double emulsion and
recognize cancer-specific epitopes overexpressed on tumor solvent evaporation, and emulsion polymerization.[268] An
cells and vasculature have been used as targeting moieties for oppositely charged polymer can be used to entrap drugs in the
drugs and drug carriers. For example, RGD peptides showed polymeric matrix of a nanoparticle, which then releases the
a high affinity in binding toward integrin,[260] which are drug through a combination of drug diffusion and polymer
typically overexpressed by the endothelium during tumor degradation. The double emulsion and solvent evaporation
angiogenesis. By conjugating RGD to the surface of SPIONs, techniques involve dissolution of the polymer and drug in an
the nanoparticles showed superior targeting affinity and organic solvent, followed by emulsification in an aqueous
specificity.[261] One possible disadvantage is that peptides solution. The organic solvent diffuses from the polymer phase
sometimes exhibit a lower binding affinity to receptors as to the aqueous phase, and evaporates from the aqueous phase,
compared to MAbs, but this can be compensated by increas- leaving behind drug-loaded polymer nanoparticles. The draw-
ing the coverage density of peptides. back of this method lies in the poor uniformity of the
5) Aptamers are short, single-stranded, synthetic nucleic nanoparticles that are produced. By contrast, the emulsion
acid oligomers, DNA or RNA, that can form complex three- polymerization approach is able to generate uniform, nano-
dimensional structures with a capability to bind to surface sized particles based on the polymerization of monomers in
markers with high affinity and specificity.[262] Advantages of emulsified droplets. However, only a few kinds of materials
aptamers include availability, ease of chemical synthesis, low can be used for the fabrication of nanoparticles using this
molecular weight, and lack of immunogenicity. Many publi- method.
cations have reported the conjugation of aptamers to polymer
nanoparticles as targeting ligands.[190b, 262]
6) Endogenous ligands, such as folic acid, epidermal 6. Case Studies
growth factor (EGF), and transferrin, are attractive for tumor
targeting because they can bind to their respective receptors Since the concept of nanoparticle therapeutics was
with low immunogenicity and high affinity. Several protocols conceived in 1955, many different types of carrier systems
have been reported to conjugate folic acid,[263] EGF,[264] and have been demonstrated or developed. While most of them
transferrin[265] to various types of nanoparticles. are still limited to benchtop investigations, some of them have
In summary, the choice of a targeting ligand revolves either entered into the market[269] or are currently undergoing
around numerous considerations, including availability, easi- different phases of clinical trials (Table 1). In this section, we
ness of production, diversity, affinity, protocols for conjuga- use a set of selected examples to highlight a number of such
tion, immunogenicity, and cost. All of these parameters carriers, including those based on protein–drug conjugates,
should be carefully thought over when designing nanoparti- liposomes, dendrimers, hydrogels, as well as nanoparticles
cles with a maximum targeting capacity while minimizing the made of biodegradable polymers, phase-change materials,
cost. and various inorganic materials.
5.4. Drug Loading 6.1. Protein–Drug Conjugates
Theoretically, a successful drug-delivery system based on This class of drug-delivery system is based on the direct
nanoparticles should have a high drug-loading capacity to conjugation of drug molecules to proteins for targeted drug
minimize the quantity of materials needed for administration. delivery. When an antibody is used, the system is also known
Loading of drug molecules into the nanoparticles can be as antibody–drug conjugate (ADC).[270] The linker between
achieved in two different ways: 1) incorporation at the time of the protein and the drug is often biodegradable, capable of
nanoparticle formation, and 2) absorption (as well as adsorp- setting both parts free upon appropriate stimulation. The
tion) of the drug after the formation of nanoparticles by simplicity of this system lends both pros and cons to itself. The
incubating them with a highly concentrated drug solution. biggest advantage arises from the small size (ca. 10 nm) of
The efficiency of drug loading and entrapment in a nano- such conjugates, which gives them relatively long circulation
particle is determined by the properties of both the drug half-lives,[271] and makes their extravasation into tumor sites
molecules and the carrier material. The properties of the much easier compared to nanoparticles with larger sizes, even
material include its molecular weight, polymer composition, for conjugates lacking ligands for active targeting. As for the
drug–polymer interaction, and the functional groups (e.g., shortcomings, not all drugs can be readily conjugated to
carboxy or ester) at both ends of each polymer chain.[266] A proteins, as the structural sensitivity of certain drugs may
macromolecule or protein has the greatest loading efficiency exclude them from any chemical modifications. In addition,
when the drug loading is performed at or near its isoelectric the stability of drug–protein linkers can be a matter of
point, which gives it the minimum solubility and maximum concern as the linkers tend to be rapidly degraded by
absorption. For small molecules, the use of electrostatic proteases and redox-altering agents during plasma circula-
interactions between the drug and matrix material is an tion.[271, 272] It is generally believed that protease-cleavable
effective way to increase the drug-loading efficiency.[267] linkers are more stable than disulfides or other linkers,
Ü
Ü

.
Reviews
although the stability of all of them can be tailored.[271, 273]

Furthermore, the ability to induce multiple functionality to
the conjugates is somewhat limited. Despite these disadvan-
tages, technological advancement in the development of new
protein carriers (e.g., monoclonal antibodies), drugs, and
especially linkers has greatly improved the capability of
ADCs as potent therapeutics. Newer generations of products
have been shown with greatly improved systemic stability,
where the linkers can be held in place until the conjugates
arrive at the targeted site, allowing for more precise and
controllable delivery of cytotoxic agents.[270c, 274]
One example of these advancements involves two drugs:
tumor-necrosis-factor-related apoptosis-inducing ligand
(TRAIL) and monomethyl auristatin E (MMAE, Fig-
ure 14 a).[275] TRAIL can induce programmed cell death by
interacting with the death receptors DR4 and DR5.[276]
Although this drug has been proven effective, many tumor
cells, especially breast tumors and melanomas, are resistant to
TRAIL treatment.[277] Interestingly, upon binding to tumor
cells, TRAIL is always internalized through endocytosis,
although this internalization process is independent of
TRAIL-induced apoptosis. By taking advantage of this
feature, Chen and co-workers added MMAE, a synthetic
antimitotic agent that can inhibit cell division by blocking the
polymerization of tubulin,[278] to TRAIL through a linker
consisting of valine and citrulline (vc; Figure 14 b). The linker
is highly stable in body fluids, but cleavable by cathepsin (a
protease overexpressed in several types of human cancers)[279]
upon entering tumor cells. The authors found that a conjugate
of N109C (an active mutant of TRAIL) and vcMMAE had
a greatly reduced IC50 (the half maximal inhibitory concen-
tration of a therapeutic agent) of 62.5 nm toward TRAIL-
resistant MCF-7 breast tumor cells. In comparison, the IC50
values for other controls all exceeded 1000 nm, including
S96C–vcMMAE (where S96C is an inactive mutant of
TRAIL), and the fully active N109C and TRAIL by
themselves. They further tested the in vivo biodistribution
of N109C–vcMMAE in nude mice bearing NCI-H460 human
lung tumors. Intriguingly, unlike the control based on bovine
serum albumin (BSA), which tended to accumulate in several
organs, the N109C–vcMMAE that remained in the body of
a mouse 96 h after injection stayed almost entirely in the
tumor region (Figure 14 c,d). This strong targeting capability
of N109C–vcMMAE conjugates, even in the absence of an
antibody or other ligand, can be attributed to their small size
and the EPR effect at the tumor site, and the targeting
efficiency can be further improved by conjugation with an
antibody.
Figure 14. a) Delivery of MMAE to cytoplasm through a TRAIL–
vcMMAE conjugate by targeting to the death receptor on tumor cell.
6.2. Liposomes b) Chemical structure of TRAIL-vcMMAE. The arrow indicates the
cathepsin cleavage site of the linker. c) The Cy5-labeled N109C-
Liposomes refer to spherical vesicles (typically, 50– vcMMAE and Cy5-BSA were injected into NCI-H460 tumor-bearing
500 nm in diameter) consisting of a lipid bilayer, which are nude mice through the tail vein. Saline was used as a negative control.
formed when lipids are emulsified in an aqueous medium.[280] The mice were monitored for fluorescence every 24 h using a Maestro
in vivo imaging system. d) At 96 h, the mice were sacrificed and the
The formation of liposomes is a spontaneous process enabled
organs were harvested and imaged. The pseudocolor image represents
by the interactions between water molecules and amphiphilic the spatial distribution of Cy5-labeled N109C-vcMMAE or Cy5-BSA in
lipid molecules. Upon completion, an aqueous volume the organs. Reproduced with permission from Ref. [275], copyright
becomes trapped within the core of each liposome. This 2013 Wiley-VCH.
Ü
Ü

Angewandte
gives liposomes the capability to selectively sequester solutes certain intracellular compartments (e.g., endosomes and
for encapsulation, forming the basis for drug delivery. This lysosomes), or the bilayer can be fused with the membranes
drug-delivery system was first demonstrated in the 1960s,[281] of intracellular compartments. The fusion process may not
and it represents one of the few systems that have been occur if the liposomes are functionalized with certain com-
successfully translated into the clinic.[7h] pounds to eliminate a direct contact between the bilayer of
The lipids commonly used to form liposomes include a liposome and that of a cell, prompting the need for
phosphatidylcholine-enriched phospholipids, either natural additional endosomal escape mechanisms.
(e.g., cholestoral and egg phosphatidylcholine) or synthetic In a recent study, Ping and co-workers devised a novel
(e.g., 1,2-dioleoyl-sn-glycero-3-phosphocholine, DOPC). The type of multistage pH-responsive liposomes (known as
properties of a liposome, such as permeability, surface charge HHG2C18-L) for anticancer drug delivery.[286] The key com-
and hydrodynamics, are mainly determined by the phospho- ponent was 1,5-dioctadecyl-l-glutamyl 2-histidyl-hexahydro-
lipid compositions of the bilayer. The initially devised and benzoic acid (HHG2C18), a zwitterionic oligopeptide lipid.
most commonly used method of drug loading is based on the The synthetic lipid HHG2C18 can be mixed with soy
involuntary encapsulation of hydrophilic drugs dissolved in phosphatidylcholine (SPC) and cholesterol to generate the
the aqueous medium during a fabrication process. Other smart liposomes. Under physiological pH values (7.2–8.0),
methods have also been demonstrated depending on the HHG2C18-L had a strongly negative surface charge
properties of the drugs. For example, lipophilic/amphiphilic ( 22.8 mV), which reduced opsonization and prolonged
drugs can be directly mixed with the lipids prior to liposome circulation time. Upon entering the tumor where the
fabrication, leading to the trapping of the drugs within the pH value was slightly reduced to about 6.5, the zeta potential
lipid bilayers.[282] The loading of these drugs can also be changed sharply to + 6.3 mV owing to the presence of
achieved through the use of an exchange mechanism involv- hexahydrobenzoic acid (HBA). Interestingly, a second-stage
ing organic solvents, after fabrication of the liposomes.[283] pH response could occur when the liposomes entered endo-
However, unlike hydrophilic solutes, which cannot easily pass somes and/or lysosomes (pH value in endolysosomes: 5.5–
through the lipid bilayer once encapsulated, lipophilic/ 4.5) after endocytosis. This endocytosis process led to two
amphiphilic drugs may not be efficiently retained in a lip- outcomes: 1) The imidazole group of histidine in HHG2C18
osome because they can easily diffuse
across the lipid bilayer. In some cases,
the different pH value in the interior
of liposomes can effectively proton-
ate/deprotonate a neutral drug,
making the bilayer membrane no
longer permeable to the drug.[284]
Using this approach, drugs such as
doxorubicin have been successfully
encapsulated in preformed liposomes
with high loading efficiency.[285]
Liposomes can be stabilized steri-
cally by reinforcing the bilayer with
an amphiphilic, long-chain polymer
containing PEG at one end, which can
concurrently reduce opsonization and
prolong the plasma circulation time.
Polymers with proper end groups for
conjugation with antibodies or ligands
can also be inserted into the lipid
bilayer, thus making targeted delivery
possible. Despite the apparent sim-
plicity in the functionalization, the
most interesting feature of liposomes,
which differentiates them from other
nanoparticle-based drug-delivery sys-
tems, is their mechanism of intracel-
lular delivery. As the bilayers of lip-
osomes closely mimic those of cells, Figure 15. a) Intracellular delivery of the smart liposomes (HHG2C18-L) encapsulating coumarin 6
they can be directly fused with the (C6, a green fluorescent dye) in A498 cells at different time points observed by confocal
microscopy. The late endosomes and lysosomes were stained by LysoTracker Red. b) C6 content in
plasma membrane. If they are inter-
mitochondria isolated from A498 cells incubated with C6/HHG2C18-L and C6/SPC-L for 12 h and
nalized by cells through endocytosis, 24 h. *P < 0.05. c) Antitumor efficacy against Renca xenograft tumor after intravenous administra-
the lipid bilayer will be disrupted tion of different formulations of CCI-779 (10 mg kg 1). *P < 0.05, **P < 0.01. Reproduced with
because of the acidic environment of permission from Ref. [286], copyright 2012 Wiley-VCH.
Ü
Ü

.
Reviews
facilitated proton influx to endolysosomes, resulting in an files, the use of these carriers can result in a much better
even more positive surface charge (+ 15–25 mV) and endo- control over the release of drugs compared with other
lysosomal bursting, and 2) HBA was cleaved in the environ- nanoparticle systems.[291]
ment with the low pH value, preventing charge reversion As described in one report, it is possible to attain
from positive to negative when HHG2C18-L escaped from the simultaneous delivery of siRNA and paclitaxel by polymer
endolysosomes to the cytoplasm. In addition, HHG2C18-L has nanoparticles based on a biodegradable triblock copolymer,
the ability to target mitochondria through electrostatic PEG-b-PCL-b-poly(2-aminoethylethylene phosphate)
interactions because it carries a strong, positive surface (mPEG45-b-PCL80-b-PPEEA10), for synergistic tumor sup-
charge. pression (Figure 16 a).[292] The triblock copolymer is amphi-
The ability of HHG2C18-L to respond in multiple stages philic and can self-assemble into nanoparticles with PCL as
was subsequently verified experimentally (Figure 15 a). The the hydrophobic core, PPEEA as the cationic shell, and PEG
uptake amount of HHG2C18-L by A498 human renal as the hydrophilic sheath. As a result, the hydrophobic
carcinoma cells was significantly higher at pH 6.5 than at anticancer drug paclitaxel can be readily encapsulated in the
pH 7.4 (Figure 15 b, while control liposomes without pH res- core during the formation of nanoparticles, and negatively
ponsiveness (SPC-L) did not show any apparent differences charged siRNA can be entangled with the cationic PPEEA
between the two conditions. After endocytosis by the cells, molecules in the shell. Simultaneous delivery of siRNA and
HHG2C18-L could quickly escaped from endolysosomes paclitaxel was successfully demonstrated by a high degree of
within a couple of hours, then gradually accumulated in intracellular colocalization as shown by confocal microscopy
mitochondria. In contrast, most SPC-Ls were trapped in (Figure 16 b). When loaded with polo-like kinase 1 (Plk1)-
endolysosomes, and only a small amount was accumulated in specific siRNA (siPlk1), which knocks down the overexpres-
mitochondria, even at 8 h after cellular uptake. The authors sion of the mitosis-related gene Plk1 in tumor cells, the
then encapsulated CCI-779, an inhibitor of cell proliferation, nanoparticles efficiently inhibited the expression of Plk1
within the cavities of HHG2C18-L, and used these liposomes mRNA in MDA-MB-435s human melanocytes (this cell line
carrying an anticancer drug for both in vitro and in vivo was previously described as ductal carcinomas until recent
testing. As expected, CCI-779/HHG2C18-L showed a much genetic verification[292]) in vitro in a dose-dependent manner
higher efficiency in reducing the viability of A498 cells at (Figure 16 c). These “two-in-one” nanoparticles also showed
pH 6.5 than at pH 7.4. When injected in vivo into rats bearing improvement in antitumor effect compared with those carry-
Renca (kidney renal adenocarcinoma) tumors, CCI-779/ ing only one agent and the blank control (Figure 16 d).
HHG2C18-L also exhibited better inhibition of tumor The ability to control the physical properties of polymer
growth in comparison with CCI-779/SPC-L, torisel (the nanoparticles has provided researchers with a great capacity
commercial formulation of CCI-779), or a saline control to probe the effects of different factors on the cellular delivery
(Figure 15 c). of nanoparticles. It is worth pointing out that most of the
polymer nanoparticles fabricated using emulsion and related
techniques are limited to the spherical shape, and often
6.3. Polymer Nanoparticles plagued by polydispersity in size. Recently, a technique
known as particle replication in nonwetting templates
Polymer nanoparticles are probably some of the most (PRINT)[293] was introduced to fabricate polymer nanoparti-
extensively investigated carrier systems for drug delivery. cles with uniform, controllable sizes, shapes, aspect ratios, and
Here we only discuss synthetic polymers that are hydrophobic elasticity properties. All these parameters were found to
and biodegradable (e.g., PLA, PGA, PLGA, and PCL), as affect the amount, rate, and/or pathway of cellular uptake.[294]
well as their copolymers.[7b, 59b,c, 61b, 287] The greatest advantage More systematic investigations are still needed to establish
of these synthetic polymers is that their properties, such as a comprehensive understanding of the drug-delivery system
molecular weight, hydrophobicity, biodegradability, can all be based on polymeric nanoparticles.
varied in a controllable fashion to allow for further function-
alization. The fabrication of nanoparticles from these poly-
mers is straightforward because a large number of techniques 6.4. Polymer–Lipid Hybrid Nanoparticles
have been demonstrated, including emulsification, coacerva-
tion, nanoprecipitation, and electrospray.[59b,c, 287a, 288] This While liposomes and polymer nanoparticles have been
versatility makes the encapsulation of drugs relatively easy successfully used separately, a novel class of hybrid nano-
and highly efficient. Hydrophobic drugs can be directly particles taking advantage of both systems has also been
dissolved in the solvent together with the polymer prior to developed. These hybrid nanoparticles possess high drug-
nanoparticle formation,[289] and hydrophilic drugs can be encapsulation yields, precisely controlled drug-release pro-
encapsulated using multiple methods: 1) in the same manner files, and excellent targeting capabilities. To this end, Far-
as for hydrophobic drugs, but suspended in the solvent; okhzad and co-workers reported a platform for fabricating
2) with techniques such as double emulsions to produce sub-100 nm targeted polymer–lipid hybrid nanoparticles
hollow nanoparticles with hydrophilic drugs trapped in the through a combination of self-assembly and precipitation.[295]
core;[290] and 3) being loaded onto the surface of nanoparticles As shown in Figure 17 a,b, the nanoparticles were comprised
after fabrication. Because polymer nanoparticles usually of 1) a biodegradable, hydrophobic polymer core that can be
contain dense matrices with well-defined degradation pro- loaded with water-insoluble drugs with sustained release
Ü
Ü

Angewandte
to polymer, thus the viscosity of the

PLGA solution (Figure 17 c). The
PLGA-lipid-PEG nanoparticles
showed a better sustained drug-
release profile than the cases where
the drugs were encapsulated in either
PLGA-PEG or PLGA nanoparticles
(Figure 17 d), thus indicating a high
efficacy of the lipid monolayer in
reducing the diffusivity of the drug.
These hybrid nanoparticles also
showed significantly improved tar-
geting efficiency toward prostate
cancer cells that overexpress pros-
tate-specific membrane antigen
(PSMA) after the surface of the
nanoparticles had been functional-
ized with an A10 RNA aptamer. At
the current stage of development, it
is still difficult to encapsulate large,
hydrophilic protein drugs for this
system based on hybrid nanoparti-
cles.
6.5. Dendrimers
Dendrimers are tree-like spheri-

cal macromolecules with many
branches emanating from a central
core.[296] Dendrimers are synthesized
in a shell-by-shell fashion starting
either from initiator cores (the most
commonly used divergence
approach) or from the periphery
(the less used convergence
approach), resulting in narrow poly-
Figure 16. a) Chemical structure of mPEG45-b-PCL80-b-PPEEA10 and schematic illustration of the
dispersity, together with a high level
formation of micellar nanoparticles and the loading with paclitaxel and siRNA. b) Confocal
microscopy image of intracellular distribution of Rho-paclitaxel-micelleplex-FAM-siRNA in MDA-MB- of control over both size and degree
[296b, 297]
435s cell after incubation for 2 h. c) Expression of Plk1 mRNA determined by quantitative real-time of branching. The latter fea-
PCR. *P < 0.007. d) Inhibition of MDA-MB-435s xenograft tumor growth by paclitaxel-micelleplex- ture, the control over the degree of
siPlk1 in comparison with various other formulations. Reproduced with permission from Ref. [291], branching, distinguishes dendrimers
copyright 2011 American Chemical Society. from other polymers that display
irregular, uncontrollable polymer
networks. As a result of the unique
rates, 2) a hydrophilic stealth coating on the surface to step-by-step synthesis procedure, the choice over the central
improve circulation half-life, and 3) a lipid monolayer at the core and the repeated units is of tremendous importance
interface between the core and the shell to retard the escape because it determines the molecular weight, size, branch
of drugs from the polymer core as well as the influx of water, density, flexibility, water solubility, and versatility in the final
thus prolonging the total period of release. Specifically, functionalization of dendrimers.[298] Dendrimers were first
PLGA and anti-cancer drug Dtxl were dissolved in acetoni- reported in the 1980s,[299] but not until recently have they
trile, while lecithin and 1,2-distearoyl-snglycero-3-phosphoe- begun to show promise in the field of drug delivery.[296b,c]
thanolamine-N-carboxy(PEG)2000 (DSPE-PEG) were Host–guest interactions (e.g., drug loading) can take place
codissolved in an aqueous solution of ethanol. By dropwise either in the interior or on the periphery of dendrimers to
addition of the PLGA/Dtxl solution to the preheated lipid result in different release profiles.
solution under gentle stirring, PLGA precipitated out as In one example, Minko and co-workers utilized poly(-
nanoparticles containing Dtxl, and the lipids self-assembled propylenimine tetrahexacontaamine) dendrimer generation 5
on the nanoparticles. The size and surface charge of the (PPIG5) as starting materials to condense with siRNA against
nanoparticles could be controlled by varying the ratio of lipid B-cell lymphoma (BCL, anti-apoptotic) mRNA.[300] The
Ü
Ü

.
Reviews
include chitosan, hya-

luronic acid, dextran,
alginate, collagen, and
gelatin; synthetic
polymers include
poly(2-hydroxyethyl
methacrylate)
(PHEMA), poly(2-
hydroxypropyl meth-
acrylate) (PHPMA),
PAAm, poly(vinyl
alcohol) (PVA), and
poly(ethylene oxide)
(PEO).[301b,c, 302] These
water-soluble poly-
mers are rendered
insoluble by physi-
cally or chemically
induced cross-linking.
For instance, while
chitosan is soluble in
acidic solutions, it
becomes insoluble at
Figure 17. a) Schematic illustration of the formation of polymer–lipid hybrid nanoparticles. The nanoparticles
comprised a hydrophobic poly(lactic-co-glycolic acid) (PLGA) core, a hydrophilic poly(ethylene glycol) (PEG) shell, neutral pH values;[303]
and a lipid (lecithin) monolayer at the interface of the hydrophobic core and the hydrophilic shell. b) Transmission alginate can be physi-
electron microscopy (TEM) image of the hybrid nanoparticles. c) Drug-encapsulation yield of PLGA-lipid-PEG hybrid cally cross-linked in
nanoparticles in comparison with PLGA-PEG and PLGA nanoparticles. d) Drug-release profiles for the PLGA-lipid- the presence of diva-
PEG, PLGA-PEG, and PLGA nanoparticles, respectively. Reproduced with permission from Ref. [295], copyright 2008 lent cations such as
American Chemical Society. Ca2+ and Ba2+;[304]
most natural biopoly-
mers are cross-linka-
PPIG5-siRNA complexes were coated with PEG for steric ble by carbodiimide reactions.[305] Compared with other
stabilization and then a synthetic analogue of luteinizing formulations, hydrogels possess several unique advantages.
hormone-releasing hormone (LHRH) peptide as the target- The high water content of hydrogels makes them resemble
ing motif. The PEG chains were linked to the surface of the biological tissues, thus reducing interfacial tension with
dendrimers by dimethyl-3-3’-dithiobispropionimidate-HCl biological fluids and promoting biocompatibility.[301b,c] The
(DTBP), a redox-responsive disulfide linkage, to facilitate porosity of a hydrogel can be tuned by controlling the density
intracellular release of siRNA once the PEG chains under- of cross-linking in the gel matrix, which strongly affects drug
went cleavage in the presence of reducing agents such as loading and subsequent release rates. Although most syn-
glutathione (Figure 18 a). As shown in Figure 18 b, targeted thetic hydrogels are not biodegradable in their original
LHRH-PEG-DTBP-PPIG5-siRNA showed extensive cellu- compositions, enzymatic, hydrolytic, and stimuli-responsive
lar uptake for LHRH-positive A2780 human ovarian carci- moieties can be incorporated into the networks to render
noma cells, but not for LHRH-negative SKOV-3 cells. them degradable under appropriate conditions.[306] Hydrogels
Nontargeted nanoparticles did not show significant intra- can be prepared as nanoparticles, and are accordingly termed
cellular accumulation in either of the two cell types. A similar nanogels.[307]
trend for silencing efficiency toward BCL mRNA was Wang and co-workers have developed charge-converting
observed (Figure 18 c,d). In vivo investigations further dem- nanogels that could be activated for drug-delivery applica-
onstrated that LHRH-PEG-DTBP-PPIG5-siRNA had tions by the acidity in tumors.[308] They first fabricated the
a higher tumor-targeting efficiency (for both dendrimers parent nanogel based on poly(2-aminoethyl methacrylate
and siRNA) and lower clearance by the liver and kidneys (for hydrochloride) (PAMA) with PEG-diacrylate (PEGDA) as
siRNA) than nontargeted nanoparticles (Figure 18 e,f). a cross-linker. The nanogel exhibited a uniform size of 100 nm
in water and a positive zeta potential of + 30 mV. The positive
charges on the surface of nanogels were advantageous for cell
6.6. Hydrogels uptake because they interacted strongly with the negatively
charged cell membranes. However, such charges also induce
Hydrogels are three-dimensional, cross-linked networks strong interactions with serum proteins, causing them to
of hydrophilic (water-soluble) polymers, capable of retaining aggregate and become rapidly cleared from circulation. To
water or physiological fluids in large quantities.[301] Typical solve this issue, the authors devised a similar method to the
examples of naturally occurring polymers used for hydrogels example we showed for liposomes: they added a layer of 2,3-
Ü
Ü

Angewandte
with succinic anhydride (SA), they

were not able to undergo effective
charge conversion and the zeta poten-
tial remained negative. The charge-
converting PSMA-DMMA nanogels
elicited more accumulation in tumor
cells (MDA-MB-435s) in vitro at
pH 6.8 than at pH 7.4 (Figure 19 b,c).
When loaded with doxorubicin,
PAMA-DMMA nanogels also caused
higher mortality for MDA-MA-435s
cells in vitro in a pH-dependent
manner compared with the PAMA-
SA nanogels (Figure 19 d).
6.7. Phase-Change Materials
Stimuli-responsiveness is a promis-
ing strategy for realizing on-demand
release of drugs or bioactive mole-
cules. Among various types of stimuli,
temperature variation has often been
employed to initiate drug release
because local body temperatures can
vary in response to ambient conditions
and in some cases, diseases.[309] One of
the best-known systems is based on
poly(N-isopropylacrylamide) (pNI-
PAAm) and its derivatives.[310] For
nanoparticles (or bulk gels) compris-
ing of cross-linked PNIPAAm chains,
they shrink upon elevation of the
temperature, forcing the encapsulated
drugs to enter the surrounding
medium. There are two intrinsic short-
comings associated with a pNIPAAm-
based delivery system: noticeable
cytotoxicity[310] and the existence in
an “on” state because of their inability
to completely inhibit drug diffusion.
Figure 18. a) Preparation of stable, tumor-targeted nanoparticles for siRNA delivery. b) Confocal To better use temperature varia-
microscopy images of LHRH receptor-positive A2780 and LHRH receptor-negative SKOV-3 human
tion as a stimulus for triggering drug
ovarian cancer cells incubated with fluorescent labeled, nontargeted PEG-DTBP-PPI G5-siRNA-6-
FAM Green particles and targeted LHRH-PEG-DTBP-PPI G5-siRNA-6-FAM Green particles. release, Xia and co-workers intro-
c,d) Suppression of BCL2 mRNA by different nanoparticles containing BCL2 targeted siRNA. duced phase-change materials
c) 1. control (fresh media), 2. PPI G5-siRNA nanoparticles, 3. PEG-DTBP-PPI G5-siRNA nano- (PCMs), which are capable of under-
particles, and 4. PEG-DTBP-siRNA-PPI G5 nanoparticles incubated with LHRH-positive A549 going reversible solid–liquid phase
cancer cells. d) 1. control (fresh media), A549 cancer cells; 2.–4. targeted siRNA nanoparticles transitions in response to changes in
incubated with LHRH-positive 2. A2780 and 3. A549 cancer cells, and LHRH-negative 4. SKOV-3
temperature.[311] In the solid state,
cancer cells. *P < 0.05. e,f) Average concentration per gram of organ weight of labeled e) den-
drimers or f) siRNA. *P < 0.05. Reproduced with permission from Ref. [300], copyright 2009
PCMs can effectively prevent any
Elsevier. leakage of encapsulated drugs at tem-
peratures below their melting points.
However, when heated beyond their
dimethylmaleic anhydride (DMMA) to the surface of the melting points, they exhibit a rapid phase change to the liquid
nanogels, which changed the zeta potential of the gels to state, thus releasing the payload. For PCMs based on fatty
17 mV. After incubation in an acidic environment (pH 6.8), alcohols and fatty acids, they are particularly well-suited for
the DMMA groups were gradually cleaved to convert the drug-delivery applications in vivo because of their excellent
surface charge of the PAMA nanogels from negative to biocompatibility. Notable examples include 1-tetradecanol
positive (Figure 19 a). When the PAMA nanogels were coated (melting point: 38–39 8C), tridecanoic acid (41–42 8C), and
Ü
Ü

.
Reviews
microbeads, which were then encapsulated in a 1-

tetradecanol block (or beads). The block was then
attached to the bottom of a container and immersed
in phosphate-buffered saline (PBS). Upon increas-
ing the temperature, the block started to melt and
released gelatin beads within 60 s. The PCM com-
pletely disappeared within 120 s, releasing all of the
gelatin microbeads. The release of FITC-dextran
ensued, as continuous heating resulted in acceler-
ated dissolution of gelatin microbeads. The rate of
FITC-dextran release could be modulated by judi-
cially selecting the materials used for PCM and
microbeads. As shown in Figure 20 b, gelatin
microbeads could be used to achieve a fast rate of
drug release as a result of the temperature respon-
siveness and hydrophilic nature of gelatin. In con-
trast, the release of FITC-dextran encapsulated in
chitosan microbeads showed a more sustained
profile because of the insolubility of chitosan in an
aqueous medium at neutral pH values. When PLGA
microbeads were used, the release rate was further
reduced as a result of the strong hydrophobicity of
the polymer. It was also demonstrated that dual
temperature-regulated drug release could be ach-
ieved by incorporating an additional block of
dodecanoic acid, which has a higher melting point
than 1-tetradecanol. The release of FITC-dextran
from gelatin microbeads within the diblock showed
a stepwise profile, dependent on the specific temper-
ature the block was exposed to (Figure 20 c). It
should be pointed out that although the examples
given here are on the microscale, PCMs can be
potentially scaled down to the nanoscale suited for
various drug-delivery applications.[311]
6.8. Inorganic Nanoparticles
Inorganic nanoparticles comprise a very impor-

tant category of drug-delivery systems because of
their rich variety, precision in size/shape control,
Figure 19. a) Comparison of drug release from pH-responsive PAMA-DMMA excellent physicochemical properties, and multi-
nanogels and PAMA-SA nanogels nonsensitive to pH change. b) Confocal micros- functionality, although their inability to degrade
copy images of MDA-MB-435s cells after incubation with PAMA-DMMA nanogels
has somewhat limited their scope of application. The
at pH 6.8 (left) and pH 7.4 (right) for 2 h. PAMA-DMMA was labeled with
fluorescein isothiocyanate (FITC; green); the F-actin and nuclei of the cells were most widely used inorganic nanoparticles include
stained with rhodamine phalloidin (red) and 4’,6-diamidino-2-phenylindole (DAPI; MSNs, carbon-based nanostructures (fullerenes,
blue), respectively. c) Comparison of cellular uptake of FITC-labeled PAMA-DMMA carbon nanotubes or CNTs, and graphene), noble-
at pH 7.4 and 6.8 by flow cytometry. d) Cell viability of MDA-MB-435 s cells after metal (typically Au) nanostructures, porous silicon,
incubation with doxorubicin (DOX)-loaded PAMA-DMMA, DOX-loaded PAMA-SA, and hydroxyapatite. Here we only focus on the first
and free DOX at the same DOX concentration of 16 mg mL 1. *P < 0.005.
three types of materials.
Reproduced with permission from Ref. [308a], copyright 2010 Wiley-VCH.
MSNs were developed in early 1990s[312] and
have since become some of the most successful
inorganic systems for drug delivery. MSNs are silica
dodecanoic acid (43–46 8C). In practice, the melting points of nanoparticles with sizes in the range of approximately 50–
PCMs can be precisely tuned in the range of 38–46 8C by using 300 nm, containing hundreds of empty channels (i.e., meso-
binary or tertiary mixtures of these compounds at appropriate pores) in a honeycomb-like arrangement. These nanoparticles
ratios. possess a high surface area and pore volume, a stable
As illustrated in Figure 20 a, a model drug, fluorescein mesostructure, a tunable pore diameter (ca. 2–10 nm), and
isothiocyanate (FITC)-dextran, was loaded into gelatin easily functionalizable surfaces (including the channels).
Ü
Ü

Angewandte
endocytosis.[317] The extremely high specific surface areas of

CNTs allow for efficient drug loading. CNTs also possess
excellent optical properties:[318] the absorption of NIR light by
CNTs can be used for photothermal destruction of tumor
cells,[319] while their NIR photoluminescence enables direct
imaging of the target cells and tissues.[320] Furthermore, CNTs
present distinctive resonance-enhanced Raman signatures,
unique for Raman detection with large scattering cross-
sections.[321] In one demonstration, Dai and co-workers
conjugated paclitaxel, a widely used anticancer drug, to
branched PEG chains anchored on single-walled CNTs
(SWCNTs) (Figure 21 a).[314b] When tested in vivo with mice
Figure 21. a) Schematic illustration of conjugation of paclitaxel (PTX)

Figure 20. a) Time-lapse fluorescence micrographs showing the release to single-walled carbon nanotubes (SWNT) functionalized by phospho-
of FITC-dextran from gelatin microbeads encapsulated in a 1-tetradeca- lipids with branched PEG chains. b) The administration of nanotube–
nol block. The temperature was gradually increased by adding warm PTX conjugates were shown to suppress tumor growth in a 4T1 breast
water (60 8C) under gentle stirring. b) Release profiles at 37 and 39 8C cancer mice model. Reproduced with permission from Ref. [314b],
for FITC-dextran from gelatin, chitosan, and PLGA microbeads encap- copyright 2008 American Association for Cancer Research.
sulated in the 1-tetradecanol blocks. c) Release profiles at 37, 39, and
42 8C for FITC-dextran from gelatin microbeads encapsulated in two
blocks made of 1-tetradecanol and dodecanoic acid, respectively. bearing 4T1 breast tumors (Figure 21 b), the SWCNTs-
Reproduced with permission from Ref. [309a], copyright 2010 Wiley- paclitaxel conjugate showed prolonged blood circulation,
VCH. 10 times higher drug uptake by the tumor, and higher efficacy
in suppressing tumor growth than free paclitaxel (clinical
taxol).
Additionally, MSNs exhibit good biocompatibility and little Gold nanostructures offer a relatively new system for drug
immunogenicity.[313] MSNs have been successfully utilized in delivery. Very similar to CNTs, Au nanostructures also
applications to deliver various theranostic agents, such as possess pronounced photothermal properties for direct
drugs, diagnostic probes, enzymes, and oligonucleotides.[313a, b] cancer therapy without involving an anticancer drug, in
CNTs have been used to deliver various types of addition to a variety of optical properties (e.g., fluorescence
therapeutic agents, including drugs,[314] peptides/proteins,[315] for Au nanoclusters,[322] multiphoton luminescence for Au
plasmid DNA,[315b, 316] and RNA, into target cells through nanorods and nanocages,[323] and strong optical absorptions
Ü
Ü

.
Reviews
for all of them) for diagnostics. While drugs can be conjugated Raman scattering.[328] Here we only focus on the applications
to the surface of solid Au nanostructures (e.g., nanospheres of Au nanocages in drug delivery and cancer therapeutics.
and nanorods), Au structures with hollow interiors (e.g., Figure 22 b shows the capability to encapsulate a drug within
nanoshells[324] and nanocages)[217] allow for a much more the cavities of Au nanocages.[121c] To control the release of the
efficient encapsulation of drugs within their cavities. drug, a thin layer of a thermoresponsive polymer based on
Gold nanocages represent a novel class of Au nano- PNIPAAm was attached to the surface of the nanocages
structures with thin, porous walls, and hollow interiors through the Au S linkage.[121c] Initial drug loading was
(Figure 22 a) and can be readily prepared using a galvanic accomplished by diffusion at an elevated temperature, at
replacement reaction between Ag templates and Au3+ in an which the polymer chains shrank to expose the pores in the
aqueous solution. The edge length and wall thickness of Au walls of the nanocages. Upon cooling to physiological
nanocages can be separately adjusted to tune an array of temperature, the polymer chains relaxed back to the extended
optical properties, including multiphoton luminescence and conformation and efficiently sealed the pores, preventing the
optical scattering/absorption, to the desirable wavelengths. drug molecules from escaping into the medium. When drug
Owing to these properties, Au nanocages have been actively release was desired, a laser was applied to the nanocages to
explored as contrast agents for imaging based on optical heat up the polymer coating by taking advantage of their
coherence tomography,[325] multiphoton microscopy,[323b,c, 326] strong photothermal effect. As a result, the pores on the walls
photoacoustic tomography,[326, 327] and surface-enhanced of the nanocages were opened, allowing for faster diffusion of
the encapsulated drugs (Figure 22 c).
It is worth noting that Au nanocages
themselves can be directly used for
cancer therapy without encapsulation
of any drug. Figure 22 d,e,f demon-
strates the photothermal treatment
effect of PEGylated Au nanocages in
mice bearing U87MGwtEGFR
tumors.[329] After 24 h exposure to
a diode laser, at a power density of
0.7 W cm 2 for 10 min, the metabolic
activity of the treated tumor dropped
significantly (as measured by 18FDG/
PET imaging) in comparison to
a saline control.
7. Summary and Outlook
The utilization of nanoparticles as

drug carriers promises a significant
improvement in cancer treatment.
Targeted delivery can reduce the
systemic side effects that patients
must endure under traditional chemo-
therapy by ensuring that pronounced
cytotoxic levels of the drugs are only
present at the tumor sites. Besides
targeting, nanoparticles have been
Figure 22. a) TEM images of Au nanocages whose surfaces were covered by a polymer brush of designed to release their payloads in
pNIPAAm-co-pAAm with a low critical solution temperature of 39 8C. The inset shows a magnified response to a variety of different
TEM image of the corner of such a nanocage. b) Schematic illustration showing the release of
stimuli, either those specific to the
a drug controlled by light. c) Absorption spectra of alizarin-PEG released from the copolymer-
covered Au nanocages when exposed to a pulsed NIR laser at a power density of 10 mWcm 2 for tumor microenvironment, such as
1, 2, 4, 8, and 16 min. d) Thermographs of tumor-bearing mice after the injection of Au nanocages acidic pH values and elevated secre-
(left) and saline (right) for 10 min. e) 18F-FDG PET/CT co-registered images of tumor-bearing mice tion of certain enzymes (e.g., matrix
intravenously administrated with either Au nanocages or saline, followed by exposure to a CW NIR metalloproteinases, MMPs),[330] or
laser: 1,2) nanocage-injected mouse before and after laser irradiation; and 3,4) saline-injected external ones, such as light exposure
mouse before and after laser irradiation. The white arrows indicated the tumors exposed to the and heating, among others. Nanopar-
laser at a power density of 0.7 W cm 2 for 10 min. f) A plot showing the ratios of 18F-FDG
ticles also offer multifunctionality,
standardized uptake values (SUV, *P < 0.001) for laser-treated tumor (right tumor) and nontreated
tumor (left tumor). Panels (a–c) are reproduced with permission from Ref. [121c], copyright 2009 combining both diagnostic (i.e.,
Nature Publishing Group. Panels (d–f) are reproduced with permission from Ref. [329], copyright image contrast enhancement or
2010 Wiley-VCH. molecular recognition capability)
Ü
Ü

Angewandte
and therapeutic features, a combination known as theranos- linkers.[336] The initially administered large particles can
tics. improve the targeting efficiency that is facilitated by the
In recent years, nanoparticle-based delivery systems have EPR effect of the tumor vasculature, but as soon as they enter
also started to show promise for cancer immunotherapy the tumor regions, the linkers will be degraded by tumor-
because they allow the (co)delivery of tumor-associated specific enzymes that disassemble them into small particles
immunomodulatory agents and/or antigens to dendritic with a size of less than 6 nm, which can be subsequently
cells. The activated dendritic cells would relay the “danger” eliminated from the body by kidney filtration.
information to T cells, which then initiate the continuous and Besides the efforts on nanoparticles themselves, it is of
long-term elimination of targeted tumor cells.[331] This intri- tremendous importance to develop realistic in vitro testing
guing research theme is expected to be actively and inten- platforms that can effectively evaluate the performance of
sively explored to greatly expand the versatility and potency nanoparticle-based drug-delivery systems. To date, the major-
of nanoparticles as cancer therapeutics. ity of the delivery systems work well in vitro, but fail when
Despite the impressive progress, a number of challenges they are tested in the much more complicated in vivo
still remain to be addressed on the route toward widespread microenvironment. Pronounced differences lie between in vi-
applications of nanomedicine. For example, a fundamental tro tumor models and preclinical models (i.e., small or large
hurdle lies in the development of methods for optimal drug laboratory animals), and between animals and human bodies.
loading into and release from nanoparticles. Precise engineer- Over the past decade, three-dimensional (3D) culture systems
ing of the physiochemical properties of a carrier must be based on porous scaffolds or hydrogels have gradually
performed to ensure that it can stabilize the encapsulated replaced the conventional two-dimensional (2D) cultures on
drug molecules during inert periods (e.g., shelf storage, blood plastic tissue culture plates (TCPs) in an effort to better mimic
circulation), but become activated to release the drug once the in vivo organization of tissues. More comfortingly, a novel
they have entered the tumor site. In principle, a nanoparticle- and exciting concept termed “organ-on-a-chip” was proposed
based delivery system should seamlessly integrate high drug- a couple of years ago by Ingber and co-workers based on the
loading capacity, long circulation half-life, effective targeting pioneering work conducted by their own and Schulers
capability, releasing programmability, stimuli responsiveness, group.[337] In such an approach, 3D miniaturized in vitro
and diagnostic features. human tissues/organs (e.g., liver, lung, heart, kidney, and
Oftentimes the targeting and therapeutic capabilities of blood vessels) are created from perfusion cultures on micro-
nanoparticles are rather limited because of the highly fluidic chips, and connected to each other to form a multi-
heterogeneous and complex tumor microenvironment that organ, human-mimicry platform that can be used for testing
contains a mixture of several subpopulations, including drugs and nanoparticles. Using this “organ-on-a-chip” plat-
primary cancer cells, cancer stem cells, mutated variants, form, one can conduct a more effective evaluation of the
and tumor-associated stromal cells.[332] On the other hand, nanoparticles as drug-delivery systems and thus predict their
many tumors share some characteristics of normal tissues, in vivo behaviors. Importantly, because of the “humanized”
such as secretion of MMPs and expression of folate receptors feature of the platform, it is expected that preclinical models
that also occur at many sites other than the tumors, albeit at might be eventually eliminated.
lower concentrations.[333] To tackle this issue, two schemes Given recent technical advancements along with knowl-
have been devised, which involve multiple delivery/targeting edge accumulated over the past decades, we believe that
mechanisms. Instead of aiming at only one target in the smart, targeted nanoparticles as drug carriers will revolution-
tumor, nanoparticles can be designed to simultaneously carry ize the field of cancer therapy by significantly improving both
two or more drugs with programmed release profiles, or the the quality and duration of a patients life. We hope to see, in
drug itself can be modified in such a way that it targets the near future, the development of personalized cancer
multiple antitumor moieties at the same time.[334] Similarly, therapeutics based on nanoparticles with increasingly sophis-
multiple antibodies specific to a tumor type (e.g., HER-2 and ticated designs and integrations.
MMP antibodies for some breast tumors)[335] can be con-
jugated to the surface of nanoparticle carriers to maximize the Received: March 5, 2014
probability and accuracy of their tumor recognition. Published online: && &&, &&&&
The in vivo clearance of nanoparticles is another critical
issue for consideration during their design. While most
[1] Globocan 2012, IARC 2013.
polymer-based nanoparticles can be designed to degrade (at
[2] S. D. Steichen, M. Caldorera-Moore, N. A. Peppas, Eur. J.
least to a certain degree) once they have accomplished their Pharm. Sci. 2013, 48, 416 – 427.
tasks as drug-delivery vehicles, many other systems (e.g., [3] Latest world cancer statistics, IARC 2013.
inorganic nanoparticles) cannot be readily degraded, and yet [4] American Cancer Society 2013.
they are too potent to be left aside. In these cases, it is [5] U. Aruna, R. Rajalakshmi, Y. I. Muzib, V. Vinesha, M. Sushma,
necessary to incorporate a higher level of complexity into the K. R. Vandana, N. V. Kumar, Inter. J. Inn. Pharm. Res. 2013, 4,
design of the delivery system based on its physiological 318 – 324.
[6] D. Peer, J. M. Karp, S. Hong, O. C. Farokhzad, R. Margalit, R.
relevance to both cancerous and normal tissues. One potential
Langer, Nat. Nanotechnol. 2007, 2, 751 – 760.
solution resides in multistep programmed systems in which [7] a) G. M. Whitesides, Nat. Biotechnol. 2003, 21, 1161 – 1165;
large particles (ca. 50 nm) are assembled from a number of b) D. A. LaVan, T. McGuire, R. Langer, Nat. Biotechnol. 2003,
small particles (< 6 nm in size) through the use of degradable 21, 1184 – 1191; c) M. Ferrari, Nat. Rev. Cancer 2005, 5, 161 –
Ü
Ü

.
Reviews
171; d) O. C. Farokhzad, J. M. Karp, R. Langer, Expert Opin. [23] a) G. J. Weiss, J. Chao, J. D. Neidhart, R. K. Ramanathan, D.
Drug Delivery 2006, 3, 311 – 324; e) L. Zhang, F. X. Gu, J. M. Bassett, J. A. Neidhart, C. H. Choi, W. Chow, V. Chung, S. J.
Chan, A. Z. Wang, R. Langer, O. C. Farokhzad, Clin. Pharma- Forman, E. Garmey, J. Hwang, D. L. Kalinoski, M. Koczywas, J.
col. Ther. 2008, 83, 761 – 769; f) R. Langer, Nature 1998, 392, 5 – Longmate, R. J. Melton, R. Morgan, J. Oliver, J. J. Peterkin,
10; g) R. Langer, Science 1990, 249, 1527 – 1533; h) O. C. J. L. Ryan, T. Schluep, T. W. Synold, P. Twardowski, M. E.
Farokhzad, R. Langer, ACS Nano 2009, 3, 16 – 20. Davis, Y. Yen, Invest. New Drugs 2013, 31, 986 – 1000; b) T.
[8] a) M. E. Lobatto, V. Fuster, Z. A. Fayad, W. J. M. Mulder, Nat. Schluep, J. Hwang, J. Cheng, J. D. Heidel, D. W. Bartlett, B.
Rev. Drug Discovery 2011, 10, 835 – 852; b) C. R. Thomas, D. P. Hollister, M. E. Davis, Clin. Cancer Res. 2006, 12, 1606 – 1614;
Ferris, J. H. Lee, E. Choi, M. H. Cho, E. S. Kim, J. F. Stoddart, c) T. Schluep, J. Cheng, K. T. Khin, M. E. Davis, Cancer
J. S. Shin, J. Cheon, J. I. Zink, J. Am. Chem. Soc. 2010, 132, Chemother. Pharmacol. 2006, 57, 654 – 662; d) J. Cheng, K. T.
10623 – 10625; c) J. H. Gao, B. Xu, Nano Today 2009, 4, 37 – 51. Khin, M. E. Davis, Mol. Pharm. 2004, 1, 183 – 193; e) J. J.
[9] A. Z. Wang, R. Langer, O. C. Farokhzad, Annu. Rev. Med. Cheng, K. T. Khin, G. S. Jensen, A. J. Liu, M. E. Davis,
2012, 63, 185 – 198. Bioconjugate Chem. 2003, 14, 1007 – 1017.
[10] A. J. Montero, B. Adams, C. M. Diaz-Montero, S. Gluck, Expert [24] S. K. Libutti, G. F. Paciotti, A. A. Byrnes, H. R. Alexander, Jr.,
Rev. Clin. Pharmacol. 2011, 4, 329 – 334. W. E. Gannon, M. Walker, G. D. Seidel, N. Yuldasheva, L.
[11] M. E. R. OBrien, N. Wigler, M. Inbar, R. Rosso, E. Grischke, Tamarkin, Clin. Cancer Res. 2010, 16, 6139 – 6149.
A. Santoro, R. Catane, D. G. Kieback, P. Tomczak, S. P. [25] T. M. Allena, P. R. Cullisb, Adv. Drug Delivery Rev. 2013, 65,
Ackland, F. Orlandi, L. Mellars, L. Alland, C. Tendler, Ann. 36 – 48.
Oncol. 2004, 15, 440 – 449. [26] Y. Iwase, Y. Maitani, Mol. Pharmaceutics 2011, 8, 330 – 337.
[12] a) C. E. Petre, D. P. Dittmer, Int. J. Nanomed. 2007, 2, 277 – 288; [27] S. P. Chawla, V. S. Chua, L. Fernandez, D. Quon, W. C.
b) “Kaposis sarcoma: DaunoXome approved”: AIDS Treat Blackwelder, E. M. Gordon, F. L. Hall, Mol. Ther. 2010, 18,
News. 1996, 246, 3 – 4. 435 – 441.
[13] M. S. Angst, D. R. Drover, Clin. Pharmacokinet. 2006, 45, [28] C. Mamot, R. Ritschard, A. Wicki, G. Stehle, T. Dieterle, L.
1153 – 1176. Bubendorf, C. Hilker, S. Deuster, R. Herrmann, C. Rochlitz,
[14] Y. Barenholz, J. Controlled Release 2012, 160, 117 – 134. Lancet Oncol. 2012, 13, 1234 – 1241.
[15] C. Oerlemans, W. Bult, M. Bos, G. Storm, J. F. Nijsen, W. E. [29] C. Potera, Genet. Eng. Biotechnol. News 2011, 31, 45 – 47.
Hennink, Pharm. Res. 2010, 27, 2569 – 2589. [30] S. P. Egusquiaguirre, M. Igartua, R. M. Hernndez, J. L. Pedraz,
[16] a) “FDA approves liposomal vincristine (Marqibo) for rare Clin. Transl. Oncol. 2012, 14, 83 – 93.
leukemia”: Oncology 2012, 26, 841; b) J. A. Silverman, S. R. [31] a) M. E. Davis, Mol. Pharmaceutics 2009, 6, 659 – 668; b) M. E.
Deitcher, Cancer Chemother. Pharmacol. 2013, 71, 555 – 564. Davis, J. E. Zuckerman, C. H. Choi, D. Seligson, A. Tolcher,
[17] J. Lao, J. Madani, T. Puertolas, M. lvarez, A. Hernndez, R. C. A. Alabi, Y. Yen, J. D. Heidel, A. Ribas, Nature 2010, 464,
Pazo-Cid, . Artal, A. A. Torres, J. Drug Delivery 2013, 1067 – 1070.
456409. [32] D. Lazarus, S. Kabir, S. Eliasof, Cancer Res. 2012, 72, 5643.
[18] P. A. Dinndorf, J. Gootenberg, M. H. Cohen, P. Keegan, R. [33] P. X. Zhao, D. Astruc, ChemMedChem 2012, 7, 952 – 997.
Pazdur, Oncologist 2007, 12, 991 – 998. [34] K. H. Jung, K. P. Kim, D. H. Yoon, Y. S. Hong, C. M. Choi, J. H.
[19] a) T. Okusaka, S. Okada, H. Ueno, M. Ikeda, R. Iwata, H. Ahn, D. H. Lee, J. L. Lee, M. H. Ryu, B. Y. Ryoo, H. M. Chang,
Furukawa, K. Takayasu, N. Moriyama, T. Sato, K. Sato, T. W. Kim, S. B. Kim, S. W. Kim, C. Suh, Y. K. Kang, J. Lee,
Oncology 2002, 62, 228 – 233; b) K. Greish, J. Fang, T. Inutsuka, K. S. Bae, Y. M. Kim, J. Clin. Oncol. 2012, 30, e13104.
A. Nagamitsu, H. Maeda, Clin. Pharmacokinet. 2003, 42, 1089 – [35] H. Shen, T. Sun, M. Ferrari, Cancer Gene Ther. 2012, 19, 367 –
1105; c) H. Ishii, J. Furuse, M. Nagase, Y. Maru, M. Yoshino, T. 373.
Hayashi, Jpn. J. Clin. Oncol. 2003, 33, 570 – 573. [36] R. Friedman, J. Natl. Cancer Inst. 2011, 103, 1428 – 1429.
[20] K. Kato, K. Chin, T. Yoshikawa, K. Yamaguchi, Y. Tsuji, T. [37] a) Y. Matsumura, H. Maeda, Cancer Res. 1986, 46, 6387 – 6392;
Esaki, K. Sakai, M. Kimura, T. Hamaguchi, Y. Shimada, Y. b) T. Minko, P. Kopeckova, J. Kopecek, Pharm. Res. 1999, 16,
Matsumura, R. Ikeda, Invest. New Drugs 2012, 30, 1621 – 1627. 986 – 996; c) P. Couvreur, Adv. Drug Delivery Rev. 2013, 65, 21 –
[21] a) J. Hrkach, D. Von Hoff, M. M. Ali, E. Andrianova, J. Auer, 23; d) G. Bao, S. Mitragotri, S. Tong, Annu. Rev. Biomed. Eng.
T. Campbell, D. De Witt, M. Figa, M. Figueiredo, A. Horhota, 2013, 15, 253 – 282.
S. Low, K. McDonnell, E. Peeke, B. Retnarajan, A. Sabnis, E. [38] P. Singh, A. Singh, J. Cancer Ther. Res. 2012, 1, 5.
Schnipper, J. J. Song, Y. H. Song, J. Summa, D. Tompsett, G. [39] L. M. Hilliard, R. L. Berkow, J. Watterson, E. A. Ballard, G. K.
Troiano, T. Van Geen Hoven, J. Wright, P. LoRusso, P. W. Balzer, C. L. Moertel, Med. Pediatr. Oncol. 1997, 28, 310 – 313.
Kantoff, N. H. Bander, C. Sweeney, O. C. Farokhzad, R. [40] B. Damascelli, G. Cant, F. Mattavelli, P. Tamplenizza, P.
Langer, S. Zale, Sci. Transl. Med. 2012, 4, 128ra139; b) P. R. Bidoli, E. Leo, F. Dosio, A. M. Cerrotta, G. Di Tolla, L. F.
Gil, D. Huhn, L. L. del Mercato, D. Sasse, W. J. Parak, Frigerio, F. Garbagnati, R. Lanocita, A. Marchian, G. Patelli,
Pharmacol. Res. 2010, 62, 115 – 125; c) J. M. Chan, L. F. C. Spreafico, V. Tich, V. Vespro, F. Zunino, Cancer 2001, 92,
Zhang, R. Tong, D. Ghosh, W. W. Gao, G. Liao, K. P. Yuet, D. 2592 – 2602.
Gray, J. W. Rhee, J. J. Cheng, G. Golomb, P. Libby, R. Langer, [41] N. Ferrara, K. J. Hillan, H.-P. Gerber, W. Novotny, Nat. Rev.
O. C. Farokhzad, Proc. Natl. Acad. Sci. USA 2010, 107, 2213 – Drug Discovery 2004, 3, 391 – 400.
2218; d) S. Dhar, F. X. Gu, R. Langer, O. C. Farokhzad, S. J. [42] C. G. Foerster, C. Cursiefen, F. E. Kruse, Cornea 2008, 27, 612 –
Lippard, Proc. Natl. Acad. Sci. USA 2008, 105, 17356 – 17361; 614.
e) J. Cheng, B. A. Teply, I. Sherifi, J. Sung, G. Luther, F. X. Gu, [43] G. Xu, H. L. McLeod, Clin. Cancer Res. 2001, 7, 3314 – 3324.
E. Levy-Nissenbaum, A. F. Radovic-Moreno, R. Langer, O. C. [44] L. Toschi, G. Finocchiaro, S. Bartolini, V. Gioia, F. Cappuzzo,
Farokhzad, Biomaterials 2007, 28, 869 – 876. Future Oncol. 2005, 1, 7 – 17.
[22] a) J. L. Lee, J. H. Ahn, S. H. Park, H. Y. Lim, J. H. Kwon, S. [45] A. Otte, C. van de Wiele, R. A. Dierckx, Nucl. Med. Commun.
Ahn, C. Song, J. H. Hong, C. S. Kim, H. Ahn, Invest. New Drugs 2009, 30, 5 – 15.
2012, 30, 1984 – 1990; b) D. W. Kim, S. Y. Kim, H. K. Kim, S. W. [46] U. K. Narta, S. S. Kanwar, W. Azmi, Crit. Rev. Oncol. Hematol.
Kim, S. W. Shin, J. S. Kim, K. Park, M. Y. Lee, D. S. Heo, Ann. 2007, 61, 208 – 221.
Oncol. 2007, 18, 2009 – 2014; c) M. W. Saif, JOP 2007, 8, 166 – [47] L. Saltz, C. Easley, P. Kirkpatrick, Nat. Rev. Drug Discovery
176. 2006, 5, 987 – 988.
Ü
Ü

Angewandte
[48] T. E. Witzig, A. M. Vukov, T. M. Habermann, S. Geyer, P. J. [67] T. M. Sun, J. Z. Du, L. F. Yan, H. Q. Mao, J. Wang, Biomaterials
Kurtin, W. R. Friedenberg, W. L. White, H. I. Chalchal, P. J. 2008, 29, 4348 – 4355.
Flynn, T. R. Fitch, D. A. Welker, J. Clin. Oncol. 2005, 23, 1103 – [68] W. H. De Jong, P. J. A. Borm, Int. J. Nanomed. 2008, 3, 133 –
1108. 149.
[49] J. M. Vose, Oncologist 2004, 9, 160 – 172. [69] a) W. S. Seferos, D. A. Giljohann, H. D. Hill, A. E. Prigodich,
[50] R. Nahta, F. J. Esteva, Cancer Lett. 2006, 232, 123 – 138. C. A. Mirkin, J. Am. Chem. Soc. 2007, 129, 15477 – 15479;
[51] S. C. Owen, A. K. Doak, P. Wassam, M. S. Shoichet, B. K. b) J. I. Cutler, K. Zhang, D. Zheng, E. Auyeung, A. E.
Shoichet, ACS Chem. Biol. 2012, 7, 1429 – 1435. Prigodich, C. A. Mirkin, J. Am. Chem. Soc. 2011, 133, 9254 –
[52] A. Kumari, S. K. Yadav, S. C. Yadav, Colloids Surf. B 2010, 75, 9257; c) J. I. Culter, E. Auyeung, C. A. Mirkin, J. Am. Chem.
1 – 18. Soc. 2012, 134, 1376 – 1391.
[53] J. Liu, H. Lee, M. Huesca, A. Young, C. Allen, Cancer [70] a) R. Duncan, Nat. Rev. Drug Discovery 2003, 2, 347 – 360;
Chemother. Pharmacol. 2006, 58, 306 – 318. b) R. Duncan, Nat. Rev. Cancer 2006, 6, 688 – 701; c) A. O.
[54] K. M. Huh, S. C. Lee, Y. W. Cho, J. Lee, J. H. Jeong, K. Park, J. Elzoghby, W. M. Samy, N. A. Elgindy, J. Controlled Release
Controlled Release 2005, 101, 59 – 68. 2012, 157, 168 – 182; d) E. Fleige, M. A. Quadir, R. Haag, Adv.
[55] H. K. Makadia, S. J. Siegel, Polymer 2011, 3, 1377 – 1397. Drug Delivery Rev. 2012, 64, 866 – 884; e) S. Mura, J. Nicolas, P.
[56] V. P. Torchilin, A. N. Lukyanov, Z. Gao, B. Papahadjopoulos- Couvreur, Nat. Mater. 2013, 12, 991 – 1003; f) Y. C. Wang, M. S.
Sternberg, Proc. Natl. Acad. Sci. USA 2003, 100, 6039 – 6044. Shim, H. W. Sung, Y. Xia, Adv. Funct. Mater. 2014, 24, 4206 –
[57] E. Fattal, A. Bochota, Int. J. Pharm. 2008, 364, 237 – 248. 4220.
[58] a) Y. P. Li, Y. Y. Pei, X. Y. Zhang, Z. H. Gu, Z. H. Zhou, W. F. [71] a) K. E. Uhrich, S. M. Cannizzaro, R. Langer, Chem. Rev. 1999,
Yuan, J. J. Zhou, J. H. Zhu, X. J. Gao, J. Controlled Release 99, 3181 – 3198; b) A. A. Robitzki, R. Kurz, Handb. Exp.
2001, 71, 203 – 211; b) E. Fattal, G. Barratt, Br. J. Pharmacol. Pharmacol. 2010, 197, 87 – 112.
2009, 157, 179 – 194; c) M. George, T. E. Abraham, J. Con- [72] a) R. Langer, N. Peppas, Biomaterials 1981, 2, 201 – 214;
trolled Release 2006, 114, 1 – 14; d) C. Bouclier, L. Moine, H. b) D. Y. Arifin, L. Y. Lee, C. H. Wang, Adv. Drug Delivery
Hillaireau, V. Marsaud, E. Connault, P. Opolon, P. Couvreur, E. Rev. 2006, 58, 1274 – 1325; c) J. Siepmann, F. Siepmann, Int. J.
Fattal, J. M. Renoir, Biomacromolecules 2008, 9, 2881 – 2890; Pharm. 2008, 364, 328 – 343.
e) T. Ishihara, T. Mizushima, Expert Opin. Drug Delivery 2010, [73] S. Pundir, A. Badola, D. Sharma, Int. J. Drug Res. Technol.
7, 565 – 575. 2013, 3, 12 – 20.
[59] a) Z. Chen, Trends Mol. Med. 2010, 16, 594 – 602; b) Y. C. [74] a) H. Huang, E. E. Remsen, T. Kowalewski, K. L. Wooley, J.
Wang, P. W. Li, L. X. Kong, Z. Peng, Y. Y. Luo, 2010 3rd Am. Chem. Soc. 1999, 121, 3805 – 3806; b) M. S. Wong, J. N.
International Conference on Biomedical Engineering and Cha, K. S. Choi, T. J. Deming, G. D. Stucky, Nano Lett. 2002, 2,
Informatics 2010, 1686 – 1689; c) M. Wang, K. Alberti, S. Sun, 583 – 587; c) H. P. Hentze, E. W. Kaler, Curr. Opin. Colloid
C. L. Arellano, Q. Xu, Angew. Chem. Int. Ed. 2014, 53, 2893 – Interface Sci. 2003, 8, 164 – 178; d) G. L. Li, H. Mçhwald, D. G.
2898. Shchukin, Chem. Soc. Rev. 2013, 42, 3628 – 3646.
[60] P. A. McCarron, M. Hall, Int. J. Pharm. 2008, 348, 115 – 124. [75] J. Siepmann, R. A. Siegel, M. J. Rathbone, Fundamentals and
[61] E. Leo, S. Pecquet, J. Rojas, P. Couvreur, E. Fattal, J. Micro- Applications of Controlled Release Drug Delivery, Springer,
encapsulation 1998, 15, 421 – 430. Amsterdam, 2012.
[62] R. Bodmeier, J. W. McGinity, J. Microencapsulation 1988, 5, [76] a) J. H. Gao, G. L. Liang, B. Zhang, Y. Kuang, X. X. Zhang, B.
325 – 330. Xu, J. Am. Chem. Soc. 2007, 129, 1428 – 1433; b) S. W. Cao, Y. J.
[63] a) R. Gref, Y. Minamitake, M. T. Peracchia, V. Trubetskoy, V. Zhu, M. Y. Ma, L. Li, L. Zhang, J. Phys. Chem. C 2008, 112,
Torchilin, R. Langer, Science 1994, 263, 1600 – 1603; b) R. Gref, 1851 – 1856; c) Y. Chen, H. R. Chen, L. M. Guo, Q. J. He, F.
A. Domb, P. Quellec, T. Blunk, R. H. Muller, J. M. Verbavatz, Chen, J. Zhou, J. W. Feng, J. L. Shi, ACS Nano 2010, 4, 529 –
R. Langer, Adv. Drug Delivery Rev. 2012, 64, 316 – 326; c) J. H. 539; d) S. Yang, N. Li, D. Chen, X. Qi, Y. Xu, Y. Xu, Q. Xu, H.
Lee, K. Lee, S. H. Moon, Y. Lee, T. G. Park, J. Cheon, Angew. Li, J. Lu, J. Mater. Chem. B 2013, 1, 4628 – 4636.
Chem. Int. Ed. 2009, 121, 4238 – 4243; d) N. Zheng, L. C. Yin, [77] K. Cheng, S. Peng, C. Xu, S. Sun, J. Am. Chem. Soc. 2009, 131,
Z. Y. Song, L. Ma, H. Y. Tang, N. P. Gabrielson, H. Lu, J. J. 10637 – 10644.
Cheng, Biomaterials 2014, 35, 1302 – 1314. [78] V. J. Mohanraj, Y. Chen, J. Pharm. Res. 2006, 5, 561 – 573.
[64] J. J. Shi, A. R. Votruba, O. C. Farokhzad, R. Langer, Nano Lett. [79] A. Dessy, A. M. Piras, M. Alderighi, S. Sandreschi, F. Chiellini,
2010, 10, 3223 – 3230. Nano Biomed. Eng. 2012, 4, 83 – 88.
[65] a) M. Wang, S. Sun, K. A. Alberti, Q. B. Xu, ACS Synth. Biol. [80] Y. D. Ma, Y. Zheng, X. W. Zeng, L. Q. Jiang, H. B. Chen, R. Y.
2012, 1, 403 – 407; b) J. Zhou, K. T. Shum, J. C. Burnett, J. J. Liu, L. Q. Huang, L. Mei, Int. J. Nanomed. 2011, 6, 2679 – 2688.
Rossi, Pharmaceuticals 2013, 6, 85 – 107; c) S. H. Ku, K. Kim, K. [81] a) X. Chen, C. P. Ooi, J. Biomater. Appl. 2006, 20, 287 – 302;
Choi, S. H. Kim, I. C. Kwon, Adv. Healthcare Mater. 2014, 3, b) P. Johansen, L. Moon, H. Tamber, H. P. Merkle, B. Gander,
1182 – 1193; d) Y. Ding, Z. Jiang, K. Saha, C. S. Kim, S. T. Kim, D. Sesardic, Vaccine 1999, 18, 209 – 215; c) M. Husmann, S.
R. F. Landis, V. M. Rotello, Mol. Ther. 2014, 22, 1075 – 1083. Schenderlein, M. L ck, H. Lindner, P. Kleiebudde, Int. J.
[66] a) A. Akinc, A. Zumbuehl, M. Goldberg, E. S. Leshchiner, V. Pharm. 2002, 242, 277 – 280; d) L. Lu, C. A. Garcia, A. G.
Busini, N. Hossain, S. A. Bacallado, D. N. Nguyen, J. Fuller, R. Mikos, J. Biomed. Mater. Res. 1999, 46, 236 – 244; e) M.
Alvarez, A. Borodovsky, T. Borland, R. Constien, A. de Fou- Zilberman, O. Grinber, J. Biomater. Appl. 2008, 22, 391 – 407.
gerolles, J. R. Dorkin, K. N. Jayaprakash, M. Jayaraman, M. [82] A. Gçpferich, Biomaterials 1996, 17, 103 – 114.
John, V. Koteliansky, M. Manoharan, L. Nechev, J. Qin, T. [83] T. V
limaa, S. Laaksovirta, Biomaterials 2004, 25, 1225 – 1232.
Racie, D. Raitcheva, K. G. Rajeev, D. W. Y. Sah, J. Soutschek, I. [84] S. Zuleger, B. C. Lippold, Int. J. Pharm. 2001, 217, 139 – 152.
Toudjarska, H. P. Vornlocher, T. S. Zimmermann, R. Langer, [85] J. A. Tamada, R. Langer, Proc. Natl. Acad. Sci. USA 1993, 90,
D. G. Anderson, Nat. Biotechnol. 2008, 26, 561 – 569; b) D. G. 552 – 556.
Anderson, W. D. Peng, A. Akinc, N. Hossain, A. Kohn, R. [86] A. N. F. Versypt, D. L. W. Pack, R. D. Braatz, J. Controlled
Padera, R. Langer, J. A. Sawicki, Proc. Natl. Acad. Sci. USA Release 2013, 165, 29 – 37.
2004, 101, 16028 – 16033; c) K. A. Woodrow, Y. Cu, C. J. Booth, [87] H. J. Kim, H. Matsuda, H, Zhou, I. Honma, Adv. Mater. 2006,
J. K. Saucier-Sawyer, M. J. Wood, W. M. Saltzman, Nat. Mater. 18, 3083 – 3088.
2009, 8, 526 – 533. [88] J. Kost, Clin. Mater. 1993, 13, 155 – 161.
Ü
Ü

.
Reviews
[89] a) J. Kost, R. Langer, Adv. Drug Delivery Rev. 1991, 6, 19 – 50; [106] R. de La Rica, D. Aili, M. M. Stevens, Adv. Drug Delivery Rev.
b) I. Lavon, J. Kost, J. Controlled Release 1998, 54, 1 – 7. 2012, 64, 967 – 978.
[90] a) R. Mo, T. Jiang, R. DiSanto, W. Tai, Z. Gu, Nat. Commun. [107] a) Z. Yang, G. Liang, L. Wang, B. Xu, J. Am. Chem. Soc. 2006,
2014, 5, 3364; b) B. Khorsand, G. Lapointe, C. Brett, J. K. Oh, 128, 3038 – 3043; b) Z. M. Yang, G. L. Liang, M. L. Ma, Y. Gao,
Biomacromolecules 2013, 14, 2103 – 2111; c) R. Duan, F. Xia, L. B. Xu, Small 2007, 3, 558 – 562.
Jiang, ACS Nano 2013, 7, 8344 – 8349; d) Y. Zhang, H. F. Chan, [108] a) C. Park, H. Kim, S. Kim, C. Kim, J. Am. Chem. Soc. 2009,
K. W. Leong, Adv. Drug Delivery Rev. 2013, 65, 104 – 120. 131, 16614 – 16615; b) T. H. Ku, M. P. Chien, M. P. Thompson,
[91] a) X. Zhao, J. Kim, C. A. Cezar, N. Huebsch, K. Lee, K. R. S. Sinkovits, N. H. Olson, T. S. Baker, N. C. Gianneschi, J.
Bouhadir, D. J. Mooney, Proc. Natl. Acad. Sci. USA 2011, 108, Am. Chem. Soc. 2011, 133, 8392 – 8395; c) K. Haba, M. Popkov,
67 – 72; b) R. Cheng, F. Meng, C. Deng, H. A. Klok, Z. Zhong, M. Shamis, R. A. Lerner, C. F. Barbas, D. Shabat, Angew.
Biomaterials 2013, 34, 3647 – 3657; c) S. R. Sirsi, M. A. Borden, Chem. 2005, 117, 726 – 730; Angew. Chem. Int. Ed. 2005, 44,
Adv. Drug Delivery Rev. 2013, 72, 3 – 14; d) V. Pillay, T. S. Tsai, 716 – 720.
Y. E. Choonara, L. C. du Toit, P. Kumar, G. Modi, D. Naidoo, [109] L. Zhu, P. Kate, V. P. Torchilin, ACS Nano 2012, 6, 3491 – 3498.
L. K. Tomar, C. Tyagi, V. M. K. Ndesendo, J. Biomed. Mater. [110] R. Dorresteijn, N. Billecke, M. Schwendy, S. P tz, M. Bonn,
Res. Part A 2014, 102, 2039 – 2054. S. H. Parekh, M. Klapper, K. M llen, Adv. Funct. Mater. 2014,
[92] a) K. Loomis, K. McNeeley, R. V. Bellamkonda, Soft Matter 24, 4026 – 4033.
2011, 7, 839 – 856; b) R. Lehner, P. Hunziker, Eur. J. Nanomed. [111] T. J. Harris, G. von Maltzahn, M. E. Lord, J. H. Park, A.
2012, 4, 73 – 80; c) D. Roy, J. N. Cambre, B. S. Sumerlin, Prog. Agrawal, D. H. Min, M. J. Sailor, S. N. Bhatia, Small 2008, 4,
Polym. Sci. 2010, 35, 278 – 301; d) R. Haag, F. Kratz, Angew. 1307 – 1312.
Chem. 2006, 118, 1218 – 1237; Angew. Chem. Int. Ed. 2006, 45, [112] H. Hatakeyama, H. Akita, E. Ito, Y. Hayashi, M. Oishi, Y.
1198 – 1215; e) J. Khandare, M. Caldern, N. M. Dagia, R. Nagasaki, R. Danev, K. Nagayama, N. Kaji, H. Kikuchi, Y.
Haag, Chem. Soc. Rev. 2012, 41, 2824 – 2848. Baba, H. Harashima, Biomaterials 2011, 32, 4306 – 4316.
[93] a) J. Kost, R. Langer, Adv. Drug Delivery Rev. 2001, 46, 125 – [113] A. Bernardos, L. Mondragon, E. Aznar, M. D. Marcos, R. M.
148; b) A. V. Ambade, E. N. Savariar, S. Thayumanavan, Mol. MaÇez, F. Sancenon, J. Soto, J. M. Barat, E. P. Paya, C. Guillem,
Pharmaceutics 2005, 2, 264 – 272; c) E. S. Lee, K. Na, Y. H. Bae, P. Amoros, ACS Nano 2010, 4, 6353 – 6368.
Nano Lett. 2005, 5, 325 – 329; d) D. Schmaljohann, Adv. Drug [114] C. Zhang, D. Pan, K. Luo, W. She, C. Guo, Y. Yang, Z. Gu, Adv.
Delivery Rev. 2006, 58, 1655 – 1670; e) P. Bawa, V. Pillay, Y. E. Healthcare Mater. 2014, 3, 1299 – 1308.
Choonara, L. C. du Toit, Biomed. Mater. 2009, 4, 022001. [115] a) M. H. Xiong, Y. Bao, X. Z. Yang, Y. C. Wang, B. Sun, J.
[94] a) J. O. Kim, A. V. Kabanov, T. K. Bronich, J. Controlled Wang, J. Am. Chem. Soc. 2012, 134, 4355 – 4362; b) M.
Release 2009, 138, 197 – 204; b) C. L. Lo, C. K. Huang, K. M. Tanihara, Y. Suzuki, Y. Nishimura, K. Suzuki, Y. Kakimaru,
Lin, G. H. Hsiue, Biomaterials 2007, 28, 1225 – 1235. Y. Fukunisi, J. Pharm. Sci. 1999, 88, 510 – 514.
[95] U. Borchert, U. Lipprandt, M. Bilang, A. Kimpfler, A. Rank, R. [116] T. Ta, T. M. Porter, J. Controlled Release 2013, 169, 112 – 125.
Peschka-Sűss, R. Schubert, P. Lindner, S. Főrster, Langmuir [117] a) T. Tagami, W. D. Foltz, M. J. Ernsting, C. M. Lee, I. F.
2006, 22, 5843 – 5847. Tannock, J. P. May, S. D. Li, Biomaterials 2011, 32, 6570 – 6578;
[96] M. Oishi, H. Hayashi, M. Iijima, Y. Nagasaki, J. Mater. Chem. b) Z. S. Al-Ahmady, W. T. Al-Jamal, J. V. Bossche, T. T. Bui,
2007, 17, 3720 – 3725. A. F. Drake, A. J. Mason, K. Kostarelos, ACS Nano 2012, 6,
[97] K. Na, E. S. Lee, Y. H. Bae, Bioconjugate Chem. 2007, 18, 9335 – 9346; c) K.-J. Chen, H.-F. Liang, H.-L. Chen, Y. Wang, P.-
1568 – 1574. Y. Cheng, H.-L. Liu, Y. Xia, H. W. Sung, ACS Nano 2013, 7,
[98] a) A. E. Felber, M. H. Dufresne, J. C. Leroux, Adv. Drug 438 – 446; d) K.-J. Chen, E.-Y. Chaung, S.-P. Wey, K.-J. Lin, F.
Delivery Rev. 2012, 64, 979 – 992; b) M. S. Shim, Y. J. Kwon, Cheng, C.-C. Lin, H.-L. Liu, H.-W. Tseng, C.-P. Liu, M.-C. Wei,
Biomaterials 2010, 31, 3404 – 3413; c) E. R. Gillies, J. M. J. C.-M. Liu, H.-W. Sung, ACS Nano 2014, 8, 5105 – 5115; e) M.-F.
Frchet, Chem. Commun. 2003, 1640 – 1641; d) H. Akita, R. Chung, K.-J. Chen, H.-F. Liang, Z.-X. Liao, W.-T. Chia, Y. Xia,
Ishiba, H. Hatakeyama, H. Tanaka, Y. Sato, K. Tange, M. Arai, H.-W. Sung, Angew. Chem. 2012, 124, 10236 – 10240; Angew.
K. Kubo, H. Harashima, Adv. Healthcare Mater. 2013, 2, 1120 – Chem. Int. Ed. 2012, 51, 10089 – 10093.
1125; e) S. Binauld, M. H. Stenzel, Chem. Commun. 2013, 49, [118] T. Tanaka, Phys. Rev. A 1978, 17, 763 – 766.
2082 – 2102; f) C. Du, D. Deng, L. Shan, S. Wan, J. Cao, J. Tian, [119] a) A. Hatefi, B. Amsden, J. Controlled Release 2002, 80, 9 – 28;
S. Achilefu, Y. Gu, Biomaterials 2013, 34, 3087 – 3097. b) T. Okano, Y. H. Bae, H. Jacobs, S. W. Kim, J. Controlled
[99] K. Ulbrich, T. Etrych, P. Chytil, M. Jelnkov, B. Rhov, J. Release 1990, 11, 255 – 265; c) L. Klouda, A. G. Mikos, J.
Drug Targeting 2004, 12, 477 – 489. Pharm. Biopharm. 2008, 68, 34 – 45.
[100] C. C. Lee, E. R. Gillies, M. E. Fox, S. J. Guillaudeu, J. M. J. [120] a) T. Y. Liu, S. H. Hu, D. M. Liu, S. Y. Chen, I. W. Chen, Nano
Frchet, E. E. Dy, F. C. Szoka, Proc. Natl. Acad. Sci. USA 2006, Today 2009, 4, 52 – 65; b) L. Martn, M. Alonso, A. Girotti, F. J.
103, 16649 – 16654. Arias, J. C. Rodrguez-Cabello, Biomacromolecules 2009, 10,
[101] M. Caldern, P. Welker, K. Licha, I. Fichtner, R. Graeser, R. 3015 – 3022.
Haag, F. Kratz, J. Controlled Release 2011, 151, 295 – 301. [121] a) S. Aluri, S. M. Janib, J. A. Mackay, Adv. Drug Delivery Rev.
[102] R. Tomlinson, J. Heller, S. Brocchini, R. Duncan, Bioconjugate 2009, 61, 940 – 952; b) M. Das, N. Sanson, D. Fava, E.
Chem. 2003, 14, 1096 – 1106. Kumacheva, Langmuir 2007, 23, 196 – 201; c) M. S. Yavuz, Y.
[103] a) Y. Bae, S. Fukushima, A. Harada, K. Kataoka, Angew. Cheng, J. Y. Chen, C. M. Cobley, Q. Zhang, M. Rycenga, J. Xie,
Chem. 2003, 115, 4788 – 4791; Angew. Chem. Int. Ed. 2003, 42, C. Kim, K. H. Song, A. G. Schwartz, L. V. Wang, Y. Xia, Nat.
4640 – 4643; b) Y. Bae, K. Kataoka, Adv. Drug Delivery Rev. Mater. 2009, 8, 935 – 939; d) J. Thvenot, H. Oliveira, O. Sandre,
2009, 61, 768 – 784. S. Lecommandoux, Chem. Soc. Rev. 2013, 42, 7099 – 7116;
[104] a) S. R. Van Tomme, G. Storm, W. E. Hennink, Int. J. Pharm. e) L. C. Yin, H. Y. Tang, K. H. Kim, N. Zheng, Z. Y. Song, N. P.
2008, 355, 1 – 18; b) C. A. Lorenzo, A. Concheiro, Smart Gabrielson, H. Lu, J. J. Cheng, Angew. Chem. 2013, 125, 9352 –
Materials for Drug Delivery, RSC, London, 2013; c) X. H. 9356; Angew. Chem. Int. Ed. 2013, 52, 9182 – 9186.
Xia, M. X. Yang, L. K. Oetjen, Y. Zhang, Q. G. Li, J. Y. Chen, [122] a) N. Fomina, C. McFearin, M. Sermsakdi, O. Edigin, A.
Y. Xia, Nanoscale 2011, 3, 950 – 953. Almutairi, J. Am. Chem. Soc. 2010, 132, 9540 – 9542; b) J. S.
[105] a) R. V. Ulijn, J. Mater. Chem. 2006, 16, 2217 – 2225; b) B. Law, Katz, J. A. Burdick, Macromol. Biosci. 2010, 10, 339 – 348; c) Y.
C. H. Tung, Bioconjugate Chem. 2009, 20, 1683 – 1695. Zhang, Q. Yin, L. Yin, L. Ma, L. Tang, J. Cheng, Angew. Chem.
Ü
Ü

Angewandte
2013, 125, 6563 – 6567; Angew. Chem. Int. Ed. 2013, 52, 6435 – 2959 – 2966; b) Z. Tao, B. B. Toms, J. Goodisman, T. Asefa,
6439. Chem. Res. Toxicol. 2009, 22, 1869 – 1880.
[123] a) C. Alvarez-Lorenzo, L. Bromberg, A. Concheiro, Photo- [142] B. D. Chithrani, W. C. W. Chan, Nano Lett. 2007, 7, 1542 – 1550.
chem. Photobiol. 2009, 85, 848 – 860; b) S. Sortino, J. Mater. [143] a) M. S. Cartiera, K. M. Johnson, V. Rajendran, M. J. Caplan,
Chem. 2012, 22, 301 – 318. W. M. Saltzman, Biomaterials 2009, 30, 2790 – 2798; b) L. Y. T.
[124] J. Lu, E. Choi, F. Tamanoi, J. I. Zink, Small 2008, 4, 421 – 426. Chou, K. Ming, W. C. W. Chan, Chem. Soc. Rev. 2011, 40, 233 –
[125] Q. Yuan, Y. Zhang, T. Chen, D. Lu, Z. Zhao, X. Zhang, Z. Li, 245.
C. H. Yan, W. Tan, ACS Nano 2012, 6, 6337 – 6344. [144] G. Ruan, A. Agrawal, A. I. Marcus, S. M. Nie, J. Am. Chem.
[126] H. Yan, C. Teh, S. Sreejith, L. L. Zhu, A. Kwok, W. Q. Fang, X. Soc. 2007, 129, 14759 – 14766.
Ma, K. T. Nguyen, V. Korzh, Y. L. Zhao, Angew. Chem. 2012, [145] K. Ma, D. D. Wang, Y. Lin, J. Wang, V. Petrenko, C. Mao, Adv.
124, 8498 – 8502; Angew. Chem. Int. Ed. 2012, 51, 8373 – 8377. Funct. Mater. 2013, 23, 1172 – 1181.
[127] Y. C. Liu, A. L. Le Ny, J. Schmidt, Y. Talmon, B. F. Chmelka, [146] a) F. Li, Z. P. Zhang, J. Peng, Z. Q. Cui, D. W. Pang, K. Li, H. P.
C. T. Lee, Langmuir 2009, 25, 5713 – 5724. Wei, Y. F. Zhou, J. K. Wen, X. E. Zhang, Small 2009, 5, 718 –
[128] a) A. Schroeder, M. S. Goldberg, C. Kastrup, Y. Wang, S. Jiang, 726; b) S. Acharya, S. K. Sahoo, Adv. Drug Delivery Rev. 2011,
B. J. Joseph, C. G. Levins, S. T. Kannan, R. Langer, D. G. 63, 170 – 183.
Anderson, Nano Lett. 2012, 12, 2685 – 2689; b) B. Moses, Y. [147] a) J. E. Fuller, G. T. Zugates, L. S. Ferreira, H. S. Ow, N. N.
You, Med. Chem. 2013, 3, 192 – 198; c) S. S. Agasti, A. Nguyen, U. B. Wiesner, R. S. Langer, Biomaterials 2008, 29,
Chompoosor, C. C. You, P. Ghosh, C. K. Kim, V. M. Rotello, 1526 – 1532; b) M. V. Yezhelyev, L. Qi, R. M. ORegan, S. M.
J. Am. Chem. Soc. 2009, 131, 5728 – 5729. Nie, X. H. Gao, J. Am. Chem. Soc. 2008, 130, 9006 – 9012;
[129] a) J. You, R. Zhang, C. Xiong, M. Zhong, M. Melancon, S. c) Y. H. Li, J. Wang, M. G. Wientjes, J. L. S. Au, Adv. Drug
Gupta, A. M. Nick, A. K. Sood, C. Li, Cancer Res. 2012, 72, Delivery Rev. 2012, 64, 29 – 39.
4777 – 4786; b) Z. Xiao, C. Ji, J. Shi, E. M. Pridgen, J. Frieder, J. [148] H. Duan, S. M. Nie, J. Am. Chem. Soc. 2007, 129, 3333 – 3338.
Wu, O. C. Farokhzad, Angew. Chem. 2012, 124, 12023 – 12027; [149] J. Panyam, W. Z. Zhou, S. Prabha, S. K. Sahoo, V. Labhasetwar,
Angew. Chem. Int. Ed. 2012, 51, 11853 – 11857; c) Y. T. Chang, FASEB J. 2002, 16, 1217 – 1226.
P. Y. Liao, H. S. Sheu, Y. J. Tseng, F. Y. Cheng, C. S. Yeh, Adv. [150] W. J. Li, F. Nicol, F. C. Szoka, Adv. Drug Delivery Rev. 2004, 56,
Mater. 2012, 24, 3309 – 3314; d) S. J. Leung, M. Romanowski, 967 – 985.
Theranostics 2012, 2, 1020 – 1036.
[151] Y. C. Tseng, S. Mozumdar, L. Huang, Adv. Drug Delivery Rev.
[130] S. Sharifi, S. Behzadi, S. Laurent, M. L. Forrest, P. Stroeve, M.
2009, 61, 721 – 731.
Mahmoudi, Chem. Soc. Rev. 2012, 41, 2323 – 2343.
[152] K. Avgoustakis, A. Beletsi, Z. Panagi, P. Klepetsanis, A. G.
[131] a) C. Yue, P. Liu, M. Zheng, P. Zhao, Y. Wang, Y. Ma, L. Cai,
Karydas, D. S. Ithakissios, J. Controlled Release 2002, 79, 123 –
Biomaterials 2013, 34, 6853 – 6861; b) L. Cheng, W. He, H.
135.
Gong, C. Wang, Q. Chen, Z. Cheng, Z. Liu, Adv. Funct. Mater.
[153] J. O. Martinez, C. Chiappini, A. Ziemys, A. M. Faust, M. Kojic,
2013, 23, 5893 – 5902; c) E. G. Graham, C. M. Macneill, N. H.
X. Liu, M. Ferrari, E. Tasciotti, Biomaterials 2013, 34, 8469 –
Levi-Polyachenko, Nano LIFE 2013, 3, 1330002.
8477.
[132] a) J. Yu, D. Javier, M. A. Yaseen, N. Nitin, R. Richards-Kortum,
[154] R. Krishna, L. D. Mayer, Eur. J. Cancer Sci. 2000, 11, 265 – 283.
B. Anvari, M. S. Wong, J. Am. Chem. Soc. 2010, 132, 1929 –
[155] a) Q. He, J. Shi, Adv. Mater. 2014, 26, 391 – 411; b) M.
1938; b) X. Zheng, F. Zhou, B. Wu, W. R. Chen, D. Xing, Mol.
Saraswathy, S. Gong, Biotechnol. Adv. 2013, 31, 1397 – 1407.
Pharmaceutics 2012, 9, 514 – 522; c) A. J. Gomes, L. O. Lunardi,
[156] L. E. van Vlerken, Z. Duan, M. V. Seiden, M. M. Amiji, Cancer
J. M. Marchetti, C. N. Lunardi, A. C. Tedesco, Photomed. Laser
Surg. 2006, 24, 514 – 521; d) H. Mok, H. Jeong, S. J. Kim, B. H. Res. 2007, 67, 4843 – 4850.
Chung, Chem. Commun. 2012, 48, 8628 – 8630. [157] Q. He, Y. Gao, L. Zhang, Z. Zhang, F. Gao, X, Ji, Y, Li, J. Shi,
[133] M. Zheng, C. Yue, Y. Ma, P. Gong, P. Zhao, C. Zheng, Z. Sheng, Biomaterials 2011, 32, 7711 – 7720.
P. Zhang, Z. Wang, L. Cai, ACS Nano 2013, 7, 2056 – 2067. [158] X. Zhang, S. Guo, R. Fan, M. Yu, F. Li, C. Zhu, Y. Gan,
[134] a) A. Kunzmann, B. Andersson, T. Thurnherr, H. Krug, A. Biomaterials 2012, 33, 7103 – 7114.
Scheynius, B. Fadeel, Biochim. Biophy. Acta 2011, 1810, 361 – [159] Y. Z. Zhao, D. D. Dai, C. T. Lu, L. J. Chen, M. Lin, X. T. Shen,
373; b) M. A. Dobrovolskaia, S. E. McNeil, Nat. Nanotechnol. X. K. Li, M. Zhang, X. Jiang, R. R. Jin, X. Li, H. F. Lv, L. Cai,
2007, 2, 469 – 478; c) L. M. Bareford, P. W. Swaan, Adv. Drug P. T. Huang, Cancer Lett. 2013, 330, 74 – 83.
Delivery Rev. 2007, 59, 748 – 758; d) T. G. Iversen, T. Skotland, [160] a) S. M. Nie, Nanomedicine 2010, 5, 523 – 528; b) C. Zhou, M.
K. Sandvig, Nano Today 2011, 6, 176 – 185. Long, Y. Qin, X. Sun, J. Zheng, Angew. Chem. 2011, 123, 3226 –
[135] S. D. Conner, S. L. Schmid, Nature 2003, 422, 37 – 44. 3230; Angew. Chem. Int. Ed. 2011, 50, 3168 – 3172.
[136] R. A. Petros, J. M. DeSimone, Nat. Rev. Drug Discovery 2010, [161] C. Burda, X. Chen, R. Narayanan, M. A. El-Sayed, Chem. Rev.
9, 615 – 627. 2005, 105, 1025 – 1102.
[137] a) A. M. Wen, N. F. Steinmetz, Adv. Healthc. Mater. 2014, DOI: [162] a) A. M. Derfus, W. C. W. Chan, S. N. Bhatia, Nano Lett. 2004,
10.1002/adhm.201400141; b) H. Herd, N. Daum, A. T. Jones, H. 4, 11 – 18; b) R. Hardman, Environ. Health Perspect. 2006, 114,
Huwer, H. Ghandehari, C.-M. Lehr, ACS Nano 2013, 7, 1961 – 165 – 172; c) M. C. Mancini, B. A. Kairdolf, A. M. Smith, S. M.
1973; c) C. Huang, Y. Zhang, H. Yua, H. Gao, S. Zhang, Nano Nie, J. Am. Chem. Soc. 2008, 130, 10836 – 10837; d) W. E. Smith,
Lett. 2013, 13, 4546 – 4550. J. Brownell, C. C. White, Z. Afsharinejad, J. Tsai, X. H. Hu, S. J.
[138] A. K. Gupta, M. Gupta, Biomaterials 2005, 26, 1565 – 1573. Polyak, X. H. Gao, T. J. Kavanagh, D. L. Eaton, ACS Nano
[139] a) I. Raynal, P. Prigent, S. Peyramaure, A. Najid, C. Rebuzzi, C. 2012, 6, 9475 – 9484.
Corot, Invest. Radiol. 2004, 39, 56 – 63; b) G. Fleige, F. [163] a) X. H. Gao, L. Yang, J. A. Petros, F. F. Marshall, J. W. Simons,
Seeberger, D. Laux, M. Kresse, M. Taupitz, H. Pilgrimm, C. S. M. Nie, Curr. Opin. Biotechnol. 2005, 16, 63 – 72; b) B. A.
Zimmer, Invest. Radiol. 2002, 37, 482 – 488. Kairdolf, A. M. Smith, T. H. Stokes, M. D. Wang, A. N. Young,
[140] P. C. Patel, D. A. Giljohann, W. L. Daniel, D. Zheng, A. E. S. M. Nie, Annu. Rev. Anal. Chem. 2013, 6, 143 – 162; c) C. E.
Prigodich, C. A. Mirkin, Bioconjugate Chem. 2010, 21, 2250 – Probst, P. Zrazhevskiy, V. Bagalkot, X. H. Gao, Adv. Drug
2256. Delivery Rev. 2013, 65, 703 – 718.
[141] a) T. H. Chung, S. H. Wu, M. Yao, C. W. Lu, Y. S. Lin, Y. Hung, [164] J. Zhao, V. Castranova, J. Toxicol. Environ. Health Part B 2011,
C. Y. Mou, Y. C. Chen, D. M. Huang, Biomaterials 2007, 28, 14, 593 – 632.
Ü
Ü

.
Reviews
[165] M. Longmire, P. L. Choyke, H. Kobayashi, Nanomedicine 2008, Kantoff, J. P. Richie, R. Langer, Proc. Natl. Acad. Sci. USA
3, 703 – 717. 2006, 103, 6315 – 6320.
[166] H. S. Choi, W. Liu, P. Misra, E. Tanaka, J. P. Zimmer, B. I. Ipe, [191] a) K. de Bruin, N. Ruthardt, K. von Gersdorff, R. Bausinger, E.
M. G. Bawendi, J. V. Frangioni, Nat. Biotechnol. 2007, 25, Wagner, M. Ogris, C. Brauchle, Mol. Ther. 2007, 15, 1297 –
1165 – 1170. 1305; b) A. R. Hilgenbrink, P. S. Low, J. Pharm. Sci. 2005, 94,
[167] a) R. P. Garay, R. El-Gewely, J. K. Armstrong, G. Garratty, P. 2135 – 2146; c) T. R. Daniels, T. Delgado, J. A. Rodriguez, G.
Richette, Expert Opin. Drug Delivery 2012, 9, 1319 – 1323; b) T. Helguera, M. L. Penichet, Clin. Immunol. 2006, 121, 144 – 158;
Shimizu, M. Ichihara, Y. Yoshioka, T. Ishida, S. Nakagawa, H. d) C. R. Dass, P. F. M. Choong, J. Controlled Release 2006, 113,
Kiwada, Biol. Pharm. Bull. 2012, 35, 1336 – 1342. 155 – 163.
[168] H. Kettiger, A. Schipanski, P. Wick, J. Huwyler, Int. J. [192] D. Neri, R. Bicknell, Nat. Rev. Cancer 2005, 5, 436 – 446.
Nanomed. 2013, 8, 3255 – 3269. [193] B. Ballou, L. A. Ernst, S. Andreko, T. Harper, J. A. J. Fitzpa-
[169] A. Albanese, P. S. Tang, W. C. W. Chan, Annu. Rev. Biomed. trick, A. S. Waggoner, M. P. Bruchez, Bioconjugate Chem. 2007,
Eng. 2012, 14, 1 – 16. 18, 389 – 396.
[170] J. W. Yoo, C. Elizabeth, M. Samir, Curr. Pharm. Des. 2012, 16, [194] a) R. Gref, M. L ck, P. Quellec, M. Marchand, E. Dellacherie,
2298 – 2307. S. Harnisch, T. Blunk, R. H. M ller, Colloids Surf. B 2000, 18,
[171] a) H. Maeda, Adv. Enzyme Regul. 2001, 41, 189 – 207; b) H. 301 – 313; b) T. Cedervall, I. Lynch, M. Foy, T. Bergg rd, S. C.
Maeda, H. Nakamura, J. Fang, Adv. Drug Delivery Rev. 2013, Donnelly, G. Cagney, S. Linse, K. A. Dawson, Angew. Chem.
65, 71 – 79. 2007, 119, 5856 – 5858; Angew. Chem. Int. Ed. 2007, 46, 5754 –
[172] S. D. Perrault, W. C. W. Chan, Proc. Natl. Acad. Sci. USA 2010, 5756; c) M. Lundqvist, J. Stigler, G. Elia, I. Lynch, T. Cedervall,
107, 11194 – 11199. K. A. Dawson, Proc. Natl. Acad. Sci. USA 2008, 105, 14265 –
[173] K. Matsumoto, T. Yamamoto, R. Kamata, H. Maeda, J. 14270.
Biochem. 1984, 96, 739 – 749. [195] T. Cedervall, I. Lynch, S. Lindman, T. Bergg rd, E. Thulin, H.
[174] G. Bergers, L. E. Benjamin, Nat. Rev. Cancer 2003, 3, 401 – 410. Nilsson, K. A. Dawson, S. Linse, Proc. Natl. Acad. Sci. USA
[175] H. Hashizume, P. Baluk, S. Morikawa, J. W. McLean, G. 2007, 104, 2050 – 2055.
Thurston, S. Roberge, R. K. Jain, D. M. McDonald, Am. J. [196] a) A. Gessner, A. Lieske, B. R. Paulke, R. H. M ller, Eur. J.
Pathol. 2000, 156, 1363 – 1380. Pharm. Biopharm. 2002, 54, 165 – 170; b) A. Vonarbourg, C.
[176] D. F. Baban, L. W. Seymour, Adv. Drug Delivery Rev. 1998, 34, Passirani, P. Saulnier, J. P. Benoit, Biomaterials 2006, 27, 4356 –
109 – 119. 4373; c) M. L ck, B. R. Paulke, W. Schrçder, T. Blunk, R. H.
[177] B. Haley, E. Frenkel, Urol. Oncol. Semin. Orig. Invest. 2008, 26, M ller, J. Biomed. Mater. Res. Part A 1998, 39, 478 – 485.
57 – 64. [197] S. M. Moghimi, J. Szebeni, Prog. Lipid Res. 2003, 42, 463 – 478.
[178] F. Yuan, M. Dellian, D. Fukumura, M. Leunig, D. A. Berk, V. P. [198] E. Kuntz, H.-D. Kuntz, Hepatology Principles and Practice:
Torchilin, R. K. Jain, Cancer Res. 1995, 55, 3752 – 3756. History, Morphology, Biochemistry Diagnostics, Clinic, Ther-
[179] R. K. Jain, T. Stylianopoulos, Nat. Rev. Clin. Oncol. 2010, 7, apy, Vol. 3, Springer, Berlin/Heidelberg, 2006.
653 – 664. [199] S. M. Moghimi, A. C. Hunter, J. C. Murray, Pharm. Rev. 2001,
[180] R. K. Jain, Cancer Res. 1987, 47, 3039 – 3051. 53, 283 – 318.
[181] C. H. Heldin, K. Rubin, K. Pietras, A. Ostman, Nat. Rev. [200] S. M. Moghimi, I. S. Muir, L. Illum, S. S. Davis, V. Kolb-
Cancer 2004, 4, 806 – 813. Bachofen, Biochim. Biophys. Acta Mol. Cell Res. 1993, 1179,
[182] M. Caldorera-Moore, N. Guimard, L. Shi, K. Roy, Expert Opin. 157 – 165.
Drug Delivery 2010, 7, 479 – 495. [201] W. C. Zamboni, V. Torchilin, A. K. Patri, J. Hrkach, S. Stern, R.
[183] P. Decuzzi, S. Lee, B. Bhushan, M. Ferrari, Ann. Biomed. Eng. Lee, A. Nel, N. J. Panaro, P. Grodzinski, Clin. Cancer Res. 2012,
2005, 33, 179 – 190. 18, 3229 – 3241.
[184] a) P. Decuzzi, R. Pasqualini, W. Arap, M. Ferrari, Pharm. Res. [202] S. W. Jones, R. A. Roberts, G. R. Robbins, J. L. Perry, M. P. Kai,
2009, 26, 235 – 243; b) Y. Geng, P. Dalhaimer, S. Cai, R. Tsai, M. K. Chen, T. Bo, M. E. Napier, J. P. Y. Ting, J. M. DeSimone,
Tewari, T. Minko, D. E. Discher, Nat. Nanotechnol. 2007, 2, J. E. Bear, J. Clin. Invest. 2013, 123, 3061 – 3073.
249 – 255. [203] W. M. Deen, M. J. Lazzara, B. D. Myers, Am. J. Physiol. Renal
[185] a) M. D. Blanco, C. Teijn, R. Olmo, J. M. Teijn, Targeted Physiol. 2001, 281, F579 – F596.
Nanoparticles for Cancer Therapy, 2012; b) V. Bagalkot, X. H. [204] M. Ohlson, J. Sçrensson, B. Haraldsson, Am. J. Physiol. Renal
Gao, ACS Nano 2011, 5, 8131 – 8139; c) Y. C. Wang, J. B. Xu, Physiol. 2001, 280, F396 – F405.
X. H. Xia, M. X. Yang, S. Vangveravong, J. Y. Chen, R. H. [205] J. I. Johnson, S. Decker, D. Zaharevitz, L. V. Rubinstein, J. M.
Mach, Y. Xia, Nanoscale 2012, 4, 421 – 424; d) T. M. Sun, Y. Venditti, S. Schepartz, S. Kalyandrug, M. Christian, S. Arbuck,
Wang, Y. C. Wang, J. B. Xu, X. Zhao, S. Vangveravong, R. H. M. Hollingshead, E. A. Sausville, Br. J. Cancer 2001, 84, 1424 –
Mach, Y. Xia, Adv. Healthcare Mater. 2014, 3, 1283 – 1291. 1431.
[186] A. Nori, K. D. Jensen, M. Tijerina, P. Kopeckova, J. Kopecek, [206] a) D. C. Drummond, O. Meyer, K. Hong, D. B. Kirpotin, D.
Bioconjugate Chem. 2003, 14, 44 – 50. Papahadjopoulos, Pharmacol. Rev. 1999, 51, 691 – 744; b) D. C.
[187] a) K. F. Pirollo, E. H. Chang, Trends Biotechnol. 2008, 26, 552 – Litzinger, A. M. J. Buiting, N. van Rooijen, L. Huang, Biochim.
558; b) O. C. Farokhzad, R. Langer, ACS Nano 2009, 3, 16 – 20. Biophys. Acta Biomembr. 1994, 1190, 99 – 107.
[188] a) A. Verma, O. Uzun, Y. Hu, Y. Hu, H. S. Han, N. Watson, S. [207] N. La-Beck, B. Zamboni, A. Gabizon, H. Schmeeda, M.
Chen, D. J. Irvine, F. Stellacci, Nat. Mater. 2008, 7, 588 – 595; Amantea, P. Gehrig, W. Zamboni, Cancer Chemother. Phar-
b) S. E. A. Gratton, P. A. Ropp, P. D. Pohlhaus, J. C. Luft, V. J. macol. 2012, 69, 43 – 50.
Madden, M. E. Napier, J. M. DeSimone, Proc. Natl. Acad. Sci. [208] H. Wu, R. K. Ramanathan, B. A. Zamboni, S. Strychor, S.
USA 2008, 105, 11613 – 11618. Ramalingam, R. P. Edwards, D. M. Friedland, R. G. Stoller,
[189] D. B. Kirpotin, D. C. Drummond, Y. Shao, M. R. Shalaby, K. C. P. Belani, L. J. Maruca, Y. J. Bang, W. C. Zamboni, J. Clin.
Hong, U. B. Nielsen, J. D. Marks, C. C. Benz, J. W. Park, Cancer Pharmacol. 2012, 52, 180 – 194.
Res. 2006, 66, 6732 – 6740. [209] E. C. Dreaden, A. M. Alkilany, X. Huang, C. J. Murphy, M. A.
[190] a) D. W. Bartlett, H. Su, I. J. Hildebrandt, W. A. Weber, M. E. El-Sayed, Chem. Soc. Rev. 2012, 41, 2740 – 2779.
Davis, Proc. Natl. Acad. Sci. USA 2007, 104, 15549 – 15554; [210] L. Minai, D. Yeheskely-Hayon, D. Yelin, Sci. Rep. 2013, 3, 2146.
b) O. C. Farokhzad, J. Cheng, B. A. Teply, I. Sherifi, S. Jon, P. W. [211] D. S. Kohane, R. Langer, Chem. Sci. 2010, 1, 441 – 446.
Ü
Ü

Angewandte
[212] S. Naahidi, M. Jafari, F. Edalat, K. Raymond, A. Khademhos- [234] W. Zauner, N. A. Farrow, A. M. Haines, J. Controlled Release
seini, P. Chen, J. Controlled Release 2013, 166, 182 – 194. 2001, 71, 39 – 51.
[213] a) M. Mahmoudi, H. Hofmann, B. Rothen-Rutishauser, A. [235] R. Vcha, F. J. Martinez-Veracoechea, D. Frenkel, Nano Lett.
Petri-Fink, Chem. Rev. 2012, 112, 2323 – 2338; b) S. A. Love, 2011, 11, 5391 – 5395.
M. A. Maurer-Jones, J. W. Thompson, Y. S. Lin, C. L. Haynes, [236] B. J. Reynwar, G. Illya, V. A. Harmandaris, M. M. Muller, K.
Annu. Rev. Anal. Chem. 2012, 5, 181 – 205; c) A. Elsaesser, Kremer, M. Deserno, Nature 2007, 447, 461 – 464.
C. V. Howard, Adv. Drug Delivery Rev. 2012, 64, 129 – 137. [237] K. Yang, Y. Q. Ma, Nat. Nanotechnol. 2010, 5, 579 – 583.
[214] M. Chidambaram, K. Krishnasamy, Int. J. Pharm. Sci. 2012, 2, [238] a) J. M. Harris, N. E. Martin, M. Modi, Clin. Pharmacokinet.
117 – 122. 2001, 40, 539 – 551; b) M. L. Adams, A. Lavasanifar, G. S.
[215] A. Alkilany, C. Murphy, J. Nanopart. Res. 2010, 12, 2313 – 2333. Kwon, J. Pharm. Sci. 2003, 92, 1343 – 1355.
[216] a) P. Monica, M. Dana, A. Simion, Nanotechnology 2009, 20, [239] M. Elsabahy, K. L. Wooley, Chem. Soc. Rev. 2012, 41, 2545 –
315602; b) A. M. Awwad, N. M. Salem, A. O. Abdeen, Int. J. 2561.
Ind. Chem. 2013, 4, 1 – 6; c) C. Wang, Y. Jiang, L. Chen, S. Li, G. [240] P. Couvreur, G. Barratt, E. Fattal, P. Legrand, C. Vauthier, Crit.
Li, Z. Zhang, Mater. Chem. Phys. 2009, 116, 388 – 391. Rev. Ther. Drug Carrier Syst. 2002, 19, 99 – 134.
[217] a) K. Nagpal, S. K. Singh, D. N. Mishra, Chem. Pharm. Bull. [241] a) M. E. Davis, Z. G. Chen, D. M. Shin, Nat. Rev. Drug
2010, 58, 1423 – 1430; b) R. C. Mundargi, V. R. Babu, V. Discovery 2008, 7, 771 – 782; b) J. Jang, S. Jeong, J. W. Seo,
Rangaswamy, P. Patel, T. M. Aminabhavi, J. Controlled Release M. C. Kim, E. Sim, Y. Oh, S. Nam, B. Park, J. Cheon, J. Am.
2008, 125, 193 – 209; c) E. Leo, B. Brina, F. Forni, M. A. Chem. Soc. 2011, 133, 7636 – 7639.
Vandelli, Int. J. Pharm. 2004, 278, 133 – 141. [242] a) A. Abuchowski, J. R. McCoy, N. C. Palczuk, T. van Es, F. F.
[218] A. Mahapatro, D. K. Singh, J. Nanobiotechnol. 2011, 9, 55. Davis, J. Biol. Chem. 1977, 252, 3582 – 3586; b) A. Abuchowski,
[219] a) E. C. Dreaden, A. M. Alkilany, X. Huang, C. J. Murphy, T. van Es, N. C. Palczuk, F. F. Davis, J. Biol. Chem. 1977, 252,
M. A. El-Sayed, Chem. Soc. Rev. 2012, 41, 2740 – 2779; b) X. H. 3578 – 3581.
Xia, Y. Xia, Front. Phys. 2014, 9, 378 – 384. [243] A. L. Klibanov, K. Maruyama, V. P. Torchilin, L. Huang, FEBS
[220] A. M. Schrand, M. F. Rahman, S. M. Hussain, J. J. Schlager, Lett. 1990, 268, 235 – 237.
D. A. Smith, A. F. Syed, Wiley Interdiscip. Rev. Nanomed. [244] S. W. Lee, M. H. Yun, S. W. Jeong, C. H. In, J. Y. Kim, M. H.
Nanobiotechnol. 2010, 2, 544 – 568. Seo, C. M. Pai, S. O. Kim, J. Controlled Release 2011, 155, 262 –
[221] J. M. Tam, J. O. Tam, A. Murthy, D. R. Ingram, L. L. Ma, K. 271.
Travis, K. P. Johnston, K. V. Sokolov, ACS Nano 2010, 4, 2178 – [245] J. N. Zheng, H. G. Xie, W. T. Yu, X. D. Liu, W. Y. Xie, J. Zhu,
2184. X. J. Ma, Langmuir 2010, 26, 17156 – 17164.
[222] a) A. I. Minchinton, I. F. Tannock, Nat. Rev. Cancer 2006, 6, [246] H. Du, P. Chandaroy, S. W. Hui, Biochim. Biophys. Acta
583 – 592; b) M. R. Dreher, W. Liu, C. R. Michelich, M. W. Biomembr. 1997, 1326, 236 – 248.
Dewhirst, F. Yuan, A. Chilkoti, J. Natl. Cancer Inst. 2006, 98, [247] a) R. L. Hong, C. J. Huang, Y. L. Tseng, V. F. Pang, S. T. Chen,
335 – 344. J. J. Liu, F. H. Chang, Clin. Cancer Res. 1999, 5, 3645 – 3652;
[223] a) A. E. Nel, L. Madler, D. Velegol, T. Xia, E. M. V. Hoek, P. b) J. Cui, C. Li, W. Guo, Y. Li, C. Wang, L. Zhang, L. Zhang, Y.
Somasundaran, F. Klaessig, V. Castranova, M. Thompson, Nat. Hao, Y. Wang, J. Controlled Release 2007, 118, 204 – 215.
Mater. 2009, 8, 543 – 557; b) M. A. Dobrovolskaia, S. E. McNeil, [248] H. Hatakeyama, H. Akita, H. Harashima, Adv. Drug Delivery
Nat. Nanotechnol. 2007, 2, 469 – 478. Rev. 2011, 63, 152 – 160.
[224] a) H. Jatzkewitz, Hoppe-Seylers Z. Physiol. Chem. 1954, 297, [249] J. Lehtinen, A. Magarkar, M. Stepniewski, S. Hakola, M.
149 – 156; b) H. Jatzkewitz, Z. Naturforsch. B 1955, 10, 27 – 31. Bergman, T. Rog, M. Yliperttula, A. Urtti, A. Bunker, Eur. J.
[225] a) K. O. Doh, Y. Yeo, Ther. Delivery 2013, 3, 1447 – 1456; b) F. Pharm. Sci. 2012, 46, 121 – 130.
Alexis, E. Pridgen, L. K. Molnar, O. C. Farokhzad, Mol. [250] a) K. Remaut, B. Lucas, K. Braeckmans, J. Demeester, S. C.
Pharmaceutics 2008, 5, 505 – 515. De Smedt, J. Controlled Release 2007, 117, 256 – 266; b) S.
[226] A. O. Elzoghby, J. Controlled Release 2013, 172, 1075 – 1091. Mishra, P. Webster, M. E. Davis, Eur. J. Cell Biol. 2004, 83, 97 –
[227] a) D. J. Gan, L. A. Lyon, Macromolecules 2002, 35, 9634 – 9639; 111.
b) M. A. Dobrovolskaia, D. R. Germolec, J. L. Weaver, Nat. [251] Z. Amoozgar, Y. Yeo, Wiley Interdiscip. Rev. Nanomed. Nano-
Nanotechnol. 2009, 4, 411 – 414. biotechnol. 2012, 4, 219 – 233.
[228] a) V. J. Mohanraj, Y. Chen, Trop. J. Pharm. Res. 2006, 5, 561 – [252] a) Z. Liu, Y. Jiao, Y. Wang, C. Zhou, Z. Zhang, Adv. Drug
573; b) N. Kamaly, Z. Xiao, P. M. Valencia, A. F. Radovic- Delivery Rev. 2008, 60, 1650 – 1662; b) T. Coviello, P. Matri-
Moreno, O. C. Farokhzad, Chem. Soc. Rev. 2012, 41, 2971 – cardi, F. Alhaique, Expert Opin. Drug Delivery 2006, 3, 395 –
3010; c) X. H. Xia, M. X. Yang, Y. C. Wang, Y. Q. Zheng, 404.
Q. G. Li, J. Y. Chen, Y. Xia, ACS Nano 2012, 6, 512 – 522. [253] a) C. Passirani, G. Barratt, J. P. Devissaguet, D. Labarre, Life
[229] a) L. Seymour, R. Duncan, J. Strohalm, J. Kopecek, J. Biomed. Sci. 1998, 62, 775 – 785; b) C. Passirani, G. Barratt, J. P.
Mater. Res. 1987, 21, 1341 – 1358; b) J. Panyam, V. Labhasetwar, Devissaguet, D. Labarre, Pharm. Res. 1998, 15, 1046 – 1050.
Adv. Drug Delivery Rev. 2003, 55, 329 – 347. [254] a) J. Li, M. Huo, J. Wang, J. Zhou, J. M. Mohammad, Y. Zhang,
[230] M. P. Desai, V. Labhasetwar, E. Walter, R. J. Levy, G. L. Q. Zhu, A. Y. Waddad, Q. Zhang, Biomaterials 2012, 33, 2310 –
Amidon, Pharm. Res. 1997, 14, 1568 – 1573. 2320; b) Z. X. Yuan, Z. R. Zhang, D. Zhu, X. Sun, T. Gong, J.
[231] M. P. Desai, V. Labhasetwar, G. L. Amidon, R. J. Levy, Pharm. Liu, C. T. Luan, Mol. Pharmaceutics 2009, 6, 305 – 314; c) I.
Res. 1996, 13, 1838 – 1845. Rivkin, K. Cohen, J. Koffler, D. Melikhov, D. Peer, R. Margalit,
[232] R. A. Kroll, M. A. Pagel, L. L. Muldoon, S. Roman-Goldstein, Biomaterials 2010, 31, 7106 – 7114; d) G. Jiang, K. Park, J. Kim,
S. A. Fiamengo, E. A. Neuwelt, Neurosurgery 1998, 43, 879 – K. S. Kim, E. J. Oh, H. Kang, S. E. Han, Y. K. Oh, T. G. Park,
886. S. K. Hahn, Biopolymers 2008, 89, 635 – 642.
[233] a) J. Kreuter, P. Ramge, V. Petrov, S. Hamm, S. E. Gelperina, B. [255] a) F. Shamsipour, A. H. Zarnani, R. Ghods, M. Chamankhah, F.
Engelhardt, R. Alyautdin, H. von Briesen, D. J. Begley, Pharm. Forouzesh, S. Vafaei, A. A. Bayar, M. M. Akhondi, M. A.
Res. 2003, 20, 409 – 416; b) Y. S. Yim, J. Choi, G. T. Kim, C. H. Oghabian, M. Jeddi-Tehrani, Avicenna J. Med. Biotechnol.
Kim, T. H. Shin, D. G. Kim, J. Cheon, Chem. Commun. 2012, 2009, 1, 27 – 31; b) M. Mahmoudi, S. Sant, B. Wang, S. Laurent,
48, 61 – 63. T. Sen, Adv. Drug Delivery Rev. 2011, 63, 24 – 46.
Ü
Ü

.
Reviews
[256] a) H. Tada, H. Higuchi, T. M. Wanatabe, N. Ohuchi, Cancer D. Baldwin, L. Ramakrishnan, C. L. Gray, K. Baker, W. I.
Res. 2007, 67, 1138 – 1144; b) X. Wu, H. Liu, J. Liu, K. N. Haley, Wood, Science 1997, 277, 818 – 821; c) G. Pan, J. Ni, Y. F. Wei,
J. A. Treadway, J. P. Larson, N. Ge, F. Peale, M. P. Bruchez, Nat. G. L. Yu, R. Gentz, V. M. Dixit, Science 1997, 277, 815 – 818.
Biotechnol. 2002, 21, 41 – 46. [277] a) A. D. Sanlioglu, E. Dirice, C. Aydin, N. Erin, S. Koksoy, S.
[257] J. W. Park, K. Hong, D. B. Colbern, R. Shalaby, J. Baselga, Y. Sanlioglu, BMC Cancer 2005, 5, 54; b) L. Zhang, B. Fang,
Shao, U. B. Nielsen, J. D. Marks, D. Moore, D. Papahadjopou- Cancer Gene Ther. 2005, 12, 228 – 237.
los, C. C. Benz, Clin. Cancer Res. 2002, 8, 1172 – 1181. [278] S. O. Doronina, B. E. Toki, M. Y. Torgov, B. A. Mendelsohn,
[258] J. Y. Chen, D. L. Wang, J. F. Xi, L. Au, A. Siekkinen, A. Warsen, C. G. Cerveny, D. F. Chace, R. L. DeBlanc, R. P. Gearing, T. D.
Z. Y. Li, H. Zhang, Y. N. Xia, X. D. Li, Nano Lett. 2007, 7, Bovee, C. B. Siegall, Nat. Biotechnol. 2003, 21, 778 – 784.
1318 – 1322. [279] T. Nomura, N. Katunuma, J. Med. Invest. 2005, 52, 1 – 9.
[259] a) W. Tai, R. Mahato, K. Cheng, J. Controlled Release 2010, 146, [280] a) G. Gregoriadis, Trends Biotechnol. 1995, 13, 527 – 537; b) A.
264 – 275; b) S. B. Uppada, T. Erickson, L. Wojdyla, D. N. Sharma, U. S. Sharma, Int. J. Pharm. 1997, 154, 123 – 140; c) Y.
Moravec, Z. Song, J. J. Cheng, N. Puri, Int. J. Nanomed. 2014, 9, Malam, M. Loizidou, A. M. Seifalian, Trends Pharmacol. Sci.
43 – 53. 2009, 30, 592 – 599.
[260] a) E. Ruoslahti, Annu. Rev. Cell Dev. Biol. 1996, 12, 697 – 715; [281] a) A. D. Bangham, M. M. Standish, J. C. Watkins, J. Mol. Biol.
b) R. R. Hantgan, M. C. Stahle, J. H. Connor, D. A. Horita, M. 1965, 13, 227 – 238; b) G. Gregoriadis, N. Engl. J. Med. 1976,
Rocco, M. A. McLane, S. Yakovlev, L. Medved, Protein Sci. 295, 765 – 770; c) G. Gregoriadis, N. Engl. J. Med. 1976, 295,
2006, 15, 1893 – 1906. 704 – 710.
[261] C. Zhang, M. Jugold, E. C. Woenne, T. Lammers, Cancer Res. [282] a) S. B. Kulkarni, G. V. Betageri, M. Singh, J. Microencapsula-
2007, 67, 1555 – 1562. tion 1995, 12, 229 – 246; b) T. Nii, F. Ishii, Int. J. Pharm. 2005,
[262] E. W. Orava, N. Cicmil, J. Gariepy, Biochim. Biophy. Acta 298, 198 – 205.
Biomembr. 2010, 1798, 2190 – 2200. [283] M. Yokoyama, A. Satoh, Y. Sakurai, T. Okano, Y. Matsumura,
[263] C. M. Cobley, L. Au, J. Y. Chen, Y. Xia, Expert Opin. Drug T. Kakizoe, K. Kataoka, J. Controlled Release 1998, 55, 219 –
Delivery 2010, 7, 577 – 587. 229.
[264] D. S. Lidke, P. Nagy, R. Heintzmann, D. J. Arndt-Jovin, J. N. [284] a) S. Clerc, Y. Barenholz, Biochim. Biophys. Acta Biomembr.
Post, H. E. Grecco, E. A. Jaras-Erijman, T. M. Jovin, Nat. 1995, 1240, 257 – 265; b) D. Zucker, D. Marcus, Y. Barenholz,
Biotechnol. 2004, 22, 198 – 203. A. Goldblum, J. Controlled Release 2009, 139, 73 – 80.
[265] Z. M. Qian, H. Li, H. Sun, K. Ho, Pharmacol. Rev. 2002, 54, [285] a) L. D. Mayer, L. C. L. Tai, M. B. Bally, G. N. Mitilenes, R. S.
561 – 587. Ginsberg, P. R. Cullis, Biochim. Biopyhs. Acta Biomembr. 1990,
[266] a) T. Govender, T. Riley, T. Ehtezazi, M. C. Garnett, S. Stolnik, 1025, 143 – 151; b) X. Li, D. J. Hirsh, D. Cabral-Lilly, A. Zirkel,
L. Illum, S. S. Davis, Int. J. Pharm. 2000, 199, 95 – 110; b) T. S. M. Gruner, A. S. Janoff, W. R. Perkins, Biochim. Biophys.
Govender, S. Stolnik, M. C. Garnett, L. Illum, S. S. Davis, J. Acta Biomembr. 1998, 1415, 23 – 40; c) A. Fritze, F. Hens, A.
Controlled Release 1999, 57, 171 – 185; c) J. Panyam, D. Kimpfler, R. Schubert, R. Peschka-S ss, Biochim. Biophys.
Williams, A. Dash, D. Leslie-Pelecky, V. Labhasetwar, J. Acta Biomembr. 2006, 1758, 1633 – 1640.
Pharm. Sci. 2004, 93, 1804 – 1814. [286] R. Mo, Q. Sun, J. Xue, N. Li, W. Li, C. Zhang, Q. Ping, Adv.
[267] Y. Chen, V. J. Mohanraj, J. E. Parkin, Lett. Pept. Sci. 2003, 10, Mater. 2012, 24, 3705 – 3705.
621 – 629. [287] a) K. S. Soppimath, T. M. Aminabhavi, A. R. Kulkarni, W. E.
[268] a) L. A. Dailey, N. Jekel, L. Fink, T. Gessler, T. Schmehl, M. Rudzinski, J. Controlled Release 2001, 70, 1 – 20; b) J. Panyam,
Wittmar, T. Kissel, W. Seeger, Toxicol. Appl. Pharmacol. 2006, V. Labhasetwar, Adv. Drug Delivery Rev. 2012, 64, 61 – 71;
215, 100 – 108; b) R. Pandey, A. Sharma, A. Zahoor, S. Sharma, c) X. X. Xia, Q. B. Xu, X. Hu, G. K. Qin, D. L. Kaplan,
G. K. Khuller, B. Prasad, J. Antimicrob. Chemother. 2003, 52, Biomacromolecules 2011, 12, 3844 – 3850; d) Q. Yin, F. Y.
981 – 986. Yap, L. C. Yin, L. Ma, Q. Zhou, L. W. Dobrucki, T. M. Fan,
[269] T. M. Allen, P. R. Cullis, Science 2004, 303, 1818 – 1822. R. C. Gaba, J. J. Cheng, J. Am. Chem. Soc. 2013, 135, 13620 –
[270] a) S. Ornes, Proc. Natl. Acad. Sci. USA 2013, 110, 13695; b) L. 13623.
Ducry, Antibody-Drug Conjugates, Humana Press, 2013; [288] R. A. Jain, Biomaterials 2000, 21, 2475 – 2490.
c) R. S. Zolot, S. Basu, R. P. Million, Nat. Rev. Drug Discovery [289] a) C. X. Song, V. Labhasetwar, H. Murphy, X. Qu, W. R.
2013, 12, 259 – 260. Humphrey, R. J. Shebuski, R. J. Levy, J. Controlled Release
[271] S. C. Alley, N. M. Okeley, P. D. Senter, Curr. Opin. Chem. Biol. 1997, 43, 197 – 212; b) C. Fonseca, S. Simes, R. Gaspar, J.
2010, 14, 529 – 537. Controlled Release 2002, 83, 273 – 286; c) L. Mu, S. S. Feng,
[272] P. D. Senter, Curr. Opin. Chem. Biol. 2009, 13, 235 – 244. Pharm. Res. 2003, 20, 1864 – 1872.
[273] a) S. C. Alley, D. R. Benjamin, S. C. Jeffrey, N. M. Okeley, D. L. [290] a) S. W. Choi, Y. Zhang, Y. Xia, Adv. Funct. Mater. 2009, 19,
Meyer, R. J. Sanderson, P. D. Senter, Bioconjugate Chem. 2008, 2943 – 2949; b) E. Cohen-Sela, M. Chorny, N. Koroukhov, H. D.
19, 759 – 765; b) R. J. Sanderson, M. A. Hering, S. F. James, Danenberg, G. Golomb, J. Controlled Release 2009, 133, 90 – 95.
M. M. C. Sun, S. O. Doronina, A. W. Siadak, P. D. Senter, A. F. [291] T. M. Sun, J. Z. Du, Y. D. Yao, C. Q. Mao, S. Dou, S. Y. Huang,
Wahl, Clin. Cancer Res. 2005, 11, 843 – 852; c) G. D. L. Phillips, P. Z. Zhang, K. W. Leong, E. W. Song, J. Wang, ACS Nano 2011,
G. Li, D. L. Dugger, L. M. Crocker, K. L. Parsons, E. Mai, 5, 1483 – 1494.
W. A. Bl
ttler, J. M. Lambert, R. V. J. Chari, R. J. Lutz, W. L. T. [292] a) G. Ellison, T. Klinowska, R. F. R. Westwood, E. Docter, T.
Wong, F. S. Jacobson, H. Koeppen, R. H. Schwall, S. R. French, J. C. Fox, Mol. Pathol. 2002, 55, 294 – 299; b) J. Rae, C.
Kenkare-Mitra, S. D. Spencer, M. X. Sliwkowski, Cancer Res. Creighton, J. Meck, B. Haddad, M. Johnson, Breast Cancer Res.
2008, 68, 9280 – 9290. Treat. 2007, 104, 13 – 19.
[274] B. Hughes, Nat. Rev. Drug Discovery 2010, 9, 665 – 667. [293] a) J. Xu, J. C. Luft, X. Yi, S. Tian, G. Owens, J. Wang, A.
[275] L. Q. Pan, H. B. Wang, Z. M. Xie, Z. H. Li, X. J. Tang, Y. C. Xu, Johnson, P. Berglund, J. Smith, M. E. Napier, J. M. DeSimone,
C. Zhang, H. Naranmandura, S. Q. Chen, Adv. Mater. 2013, 25, Mol. Pharm. 2013, 10, 3366 – 3374; b) J. L. Perry, K. G. Reuter,
4718 – 4722. M. P. Kai, K. P. Herlihy, S. W. Jones, J. C. Luft, M. Napier, J. E.
[276] a) G. Pan, K. ORourke, A. M. Chinnaiyan, R. Gentz, R. Bear, J. M. DeSimone, Nano Lett. 2012, 12, 5304 – 5310.
Ebner, J. Ni, V. M. Dixit, Science 1997, 276, 111 – 113; b) J. P. [294] a) J. Champion, A. Walker, S. Mitragotri, Pharm. Res. 2008, 25,
Sheridan, S. A. Marsters, R. M. Pitti, A. Gurney, M. Skubatch, 1815 – 1821; b) J. A. Champion, Y. K. Katare, S. Mitragotri,
Ü
Ü

Angewandte
Proc. Natl. Acad. Sci. USA 2007, 104, 11901 – 11904; c) S. [316] Y. Liu, D.-C. Wu, W.-D. Zhang, X. Jiang, C.-B. He, T. S. Chung,
Zhang, J. Li, G. Lykotrafitis, G. Bao, S. Suresh, Adv. Mater. S. H. Goh, K. W. Leong, Angew. Chem. 2005, 117, 4860 – 4863;
2009, 21, 419 – 424; d) J. Champion, S. Mitragotri, Pharm. Res. Angew. Chem. Int. Ed. 2005, 44, 4782 – 4785.
2009, 26, 244 – 249; e) J. L. Perry, K. P. Herlihy, M. E. Napier, [317] N. W. S. Kam, Z. Liu, H. Dai, J. Am. Chem. Soc. 2005, 127,
J. M. DeSimone, Acc. Chem. Res. 2011, 44, 990 – 998. 12492 – 12493.
[295] L. Zhang, J. M. Chan, F. X. Gu, J. W. Rhee, A. Z. Wang, A. F. [318] Z. Liu, S. Tabakman, K. Welsher, H. Dai, Nano Res. 2009, 2,
Radovic-Moreno, F. Alexis, R. Langer, O. C. Farokhzad, ACS 85 – 120.
Nano 2008, 2, 1696 – 1702. [319] N. W. S. Kam, M. OConnell, J. A. Wisdom, H. Dai, Proc. Natl.
[296] a) A. K. Patri, I. J. Majoros, J. R. Baker, Jr., Curr. Opin. Chem. Acad. Sci. USA 2005, 102, 11600 – 11605.
Biol. 2002, 6, 466 – 471; b) E. R. Gillies, J. M. J. Frchet, Drug [320] a) K. Welsher, Z. Liu, D. Daranciang, H. Dai, Nano Lett. 2008,
Discovery Today 2005, 10, 35 – 43; c) B. K. Nanjwade, H. M. 8, 586 – 590; b) A. de La Zerda, C. Zavaleta, S. Keren, S.
Bechra, G. K. Derkar, F. V. Manvi, V. K. Nanjwade, Eur. J. Vaithilingam, S. Bodapati, Z. Liu, J. Levi, B. R. Smith, T. J. Ma,
Pharm. Sci. 2009, 38, 185 – 196. O. Oralkan, Z. Cheng, X. Chen, H. Dai, B. T. Khuri-Yakub, S. S.
[297] S. Svenson, Eur. J. Pharm. Biopharm. 2009, 71, 445 – 462. Gambhir, Nat. Nanotechnol. 2008, 3, 57 – 62.
[298] Y. Cheng, J. Wang, T. Rao, X. He, T. Xu, Front. Biosci. 2008, 13, [321] A. Rao, E. Richter, S. Bandow, B. Chase, P. Eklund, K.
1447 – 1471. Williams, S. Fang, K. Subbaswamy, M. Menon, A. Thess,
[299] D. Tomalia, H. Baker, J. Dewald, M. Hall, G. Kallos, S. Martin, Science 1997, 275, 187 – 191.
J. Roeck, J. Ryder, P. Smith, Polym. J. 1985, 17, 117 – 132. [322] a) J. Zheng, C. Zhang, R. M. Dickson, Phys. Rev. Lett. 2004, 93,
[300] O. Taratula, O. B. Garbuzenko, P. Kirkpatrick, I. Pandya, R. 077402; b) C. A. J. Lin, T. Y. Yang, C. H. Lee, S. H. Huang,
Savla, V. P. Pozharov, T. Minko, J. Controlled Release 2009, 140, R. A. Sperling, M. Zanella, J. K. Li, J. L. Shen, H. H. Wang,
284 – 293. H. I. Yeh, W. J. Parak, W. H. Chang, ACS Nano 2009, 3, 395 –
[301] a) N. A. Peppas, Curr. Opin. Colloid Interface Sci. 1997, 2, 531 – 401.
537; b) M. Hamidi, A. Azadi, P. Rafiei, Adv. Drug Delivery Rev. [323] a) H. Wang, T. B. Huff, D. A. Zweifel, W. He, P. S. Low, A. Wei,
2008, 60, 1638 – 1649; c) T. R. Hoare, D. S. Kohane, Polymer J. X. Cheng, Proc. Natl. Acad. Sci. USA 2005, 102, 15752 –
2008, 49, 1993 – 2007. 15756; b) L. Au, Q. Zhang, C. M. Cobley, M. Gidding, A. G.
[302] J. K. Oh, D. I. Lee, J. M. Park, Prog. Polym. Sci. 2009, 34, 1261 – Schwartz, J. Y. Chen, Y. Xia, ACS Nano 2010, 4, 35 – 42; c) L.
1282. Tong, C. M. Cobley, J. Y. Chen, Y. Xia, J. X. Cheng, Angew.
[303] M. R. Kumar, R. A. Muzzarelli, C. Muzzarelli, H. Sashiwa, A. Chem. 2010, 122, 3563 – 3566; Angew. Chem. Int. Ed. 2010, 49,
Domb, Chem. Rev. 2004, 104, 6017 – 6084. 3485 – 3488; d) A. Srivatsan, S. V. Jenkins, M. Jeon, Z. J. Wu, C.
[304] O. Smidsrød, G. Skjak-Brk, Trends Biotechnol. 1990, 8, 71 – 78. Kim, J. Y. Chen, R. K. Pandey, Theranostics 2014, 4, 163 – 174.
[305] C. Mueller, J.-Y. Zhou, Methods Cell Sci. 1994, 16, 183 – 188. [324] J. Yang, J. Lee, J. Kang, S. J. Oh, H. J. Ko, J. H. Son, K. Lee, J. S.
[306] Y. Qiu, K. Park, Adv. Drug Delivery Rev. 2001, 53, 321 – 339. Suh, Y. M. Huh, S. Haam, Adv. Mater. 2009, 21, 4339 – 4342.
[307] a) A. V. Kabanov, S. V. Vinogradov, Angew. Chem. 2009, 121, [325] H. Cang, T. Sun, Z. Y. Li, J. Y. Chen, B. J. Wiley, Y. Xia, X. N.
5524 – 5536; Angew. Chem. Int. Ed. 2009, 48, 5418 – 5429; Li, Opt. Lett. 2005, 30, 3048 – 3050.
b) J. K. Oh, R. Drumright, D. J. Siegwart, K. Matyjaszewski, [326] Y. Zhang, Y. Wang, L. Wang, Y. Wang, X. Cai, C. Zhang, L. V.
Prog. Polym. Sci. 2008, 33, 448 – 477; c) J. K. Oh, D. J. Siegwart, Wang, Y. Xia, Theranostics 2013, 3, 532 – 543.
H. Lee, G. Sherwood, L. Peteanu, J. O. Hollinger, K. Kataoka, [327] C. Kim, E. C. Cho, J. Y. Chen, K. H. Song, L. Au, C. Favazza, Q.
K. Matyjaszewski, J. Am. Chem. Soc. 2007, 129, 5939 – 5945; Zhang, C. M. Cobley, F. Gao, Y. Xia, L. V. Wang, ACS Nano
d) S. V. Vinogradov, T. K. Bronich, A. V. Kabanov, Adv. Drug 2010, 4, 4559 – 4564.
Delivery Rev. 2002, 54, 135 – 147. [328] M. Rycenga, K. K. Hou, C. M. Cobley, A. G. Schwartz, P. H. C.
[308] J. Z. Du, T. M. Sun, W. J. Song, J. Wu, J. Wang, Angew. Chem. Camargo, Y. Xia, Phys. Chem. Chem. Phys. 2009, 11, 5903 –
2010, 122, 3703 – 3708; Angew. Chem. Int. Ed. 2010, 49, 3621 – 5908.
3626. [329] J. Y. Chen, C. Glaus, R. Laforest, Q. Zhang, M. X. Yang, M.
[309] a) S. W. Choi, Y. Zhang, Y. Xia, Angew. Chem. 2010, 122, 8076 – Gidding, M. J. Welch, Y. Xia, Small 2010, 6, 811 – 817.
8080; Angew. Chem. Int. Ed. 2010, 49, 7904 – 7908; b) D. C. [330] a) M. Egeblad, Z. Werb, Nat. Rev. Cancer 2002, 2, 161 – 174;
Hyun, N. S. Levinson, U. Jeong, Y. Xia, Angew. Chem. 2014, b) C. Gialeli, A. D. Theocharis, N. K. Karamanos, FEBS J.
126, 3854 – 3871; Angew. Chem. Int. Ed. 2014, 53, 3780 – 3795. 2011, 278, 16 – 27.
[310] H. Vihola, A. Laukkanen, L. Valtola, H. Tenhu, J. Hirvonen, [331] a) J. Kim, D. J. Mooney, Nano Today 2011, 6, 466 – 477; b) A. V.
Biomaterials 2005, 26, 3055 – 3064. Li, J. J. Moon, W. Abraham, H. Suh, J. Elkhader, M. A.
[311] G. D. Moon, S. W. Choi, X. Cai, W. Li, E. C. Cho, U. Jeong, Seidman, M. Yen, E.-J. Im, M. H. Foley, D. H. Barouch, D. J.
L. V. Wang, Y. Xia, J. Am. Chem. Soc. 2011, 133, 4762 – 4765. Irvine, Sci. Transl. Med. 2013, 5, 204ra130.
[312] C. Kresge, M. Leonowicz, W. Roth, J. Vartuli, J. Beck, Nature [332] T. A. Denison, Y. H. Bae, J. Controlled Release 2012, 164, 187 –
1992, 359, 710 – 712. 191.
[313] a) S. Giri, B. G. Trewyn, V. S. Lin, Nanomedicine 2007, 2, 99 – [333] S. D. Weitman, R. H. Lark, L. R. Coney, D. W. Fort, V. Frasca,
111; b) I. I. Slowing, B. G. Trewyn, S. Giri, V. Y. Lin, Adv. Funct. V. R. Zurawski, B. A. Kamen, Cancer Res. 1992, 52, 3396 – 3401.
Mater. 2007, 17, 1225 – 1236; c) I. I. Slowing, J. L. Vivero- [334] a) P. Csermely, V. Agoston, S. Pongor, Trends Pharmacol. Sci.
Escoto, C. W. Wu, V. S. Y. Lin, Adv. Drug Delivery Rev. 2008, 2005, 26, 178 – 182; b) S. Giordano, A. Petrelli, Curr. Med.
60, 1278 – 1288; d) J. L. Vivero-Escoto, I. I. Slowing, B. G. Chem. 2008, 15, 422 – 432; c) J. J. Lu, W. Pan, Y. J. Hu, Y. T.
Trewyn, V. S. Y. Lin, Small 2010, 6, 1952 – 1967. Wang, PLoS One 2012, 7, e40262.
[314] a) R. P. Feazell, N. Nakayama-Ratchford, H. Dai, S. J. Lippard, [335] a) C. M. Perou, T. Sorlie, M. B. Eisen, M. van de Rijn, S. S.
J. Am. Chem. Soc. 2007, 129, 8438 – 8439; b) Z. Liu, K. Chen, C. Jeffrey, C. A. Rees, J. R. Pollack, D. T. Ross, H. Johnsen, L. A.
Davis, S. Sherlock, Q. Cao, X. Chen, H. Dai, Cancer Res. 2008, Akslen, O. Fluge, A. Pergamenschikov, C. Williams, S. X. Zhu,
68, 6652 – 6660. P. E. Lonning, A. Borresen-Dale, P. O. Brown, D. Botstein,
[315] a) N. W. S. Kam, H. Dai, J. Am. Chem. Soc. 2005, 127, 6021 – Nature 2000, 406, 747 – 752; b) A. Scorilas, A. Karameris, N.
6026; b) A. Bianco, K. Kostarelos, M. Prato, Curr. Opin. Chem. Arnogiannaki, A. Ardavanis, P. Bassilopoulos, T. Trangas, M.
Biol. 2005, 9, 674 – 679. Talieri, Br. J. Cancer 2001, 84, 1488 – 1496; c) L. Nakopoulou, I.
Ü
Ü

.
Reviews
Tsirmpa, P. Alexandrou, A. Louvrou, C. Ampela, S. Markaki, Huh, G. A. Hamilton, D. E. Ingber, Trends Cell Biol. 2011, 12,
P. S. Davaris, Breast Cancer Res. Treat. 2003, 77, 145 – 155. 745 – 754; c) D. Huh, Y. S. Torisawa, G. A. Hamilton, H. J. Kim,
[336] L. Y. T. Chou, K. Zagorovsky, W. C. W. Chan, Nat. Nano- D. E. Ingber, Lab Chip 2012, 21, 2156 – 2164; d) J. H. Sung,
technol. 2014, 9, 148 – 155. M. L. Shuler, Lab Chip 2009, 9, 1385 – 1394; e) J. H. Sung, M. B.
[337] a) D. Huh, B. D. Matthews, A. Mammoto, M. Montoya-Zavala, Esch, M. L. Shuler, Expert Opin. Drug Metab. Toxicol. 2010, 6,
H. Y. Hsin, D. E. Ingber, Science 2010, 328, 1662 – 1668; b) D. 1063 – 1081.
Ü
Ü

Angewandte
Chemie
Reviews
Nanomedicine On the way to nanomedicine: Consider-
able advances in the development of
T. Sun, Y. S. Zhang, B. Pang, D. C. Hyun, nanoparticles for cancer therapy have
M. Yang, Y. Xia* &&&&—&&&& been made in recent years. Nanoparticle-
based drug-delivery systems offer advan-
Engineered Nanoparticles for Drug tages with regard to multidrug resistance,
Delivery in Cancer Therapy systemic delivery, and clearance, and
enable for example specific tumor tar-
geting and controlled release of thera-
peutic agents.
Ü
Ü

Engineered Nanoparticles For Drug Delivery in Cancer

Uploaded by

Copyright:

Available Formats

Engineered Nanoparticles For Drug Delivery in Cancer

Uploaded by

Document Information

Original Title

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

Engineered Nanoparticles For Drug Delivery in Cancer

Uploaded by

Copyright:

Available Formats

Angewandte

Engineered Nanoparticles for Drug Delivery in Cancer

These are not the final page numbers!

These are not the final page numbers!

These are not the final page numbers!

These are not the final page numbers!

Table 1: Nanoparticle-based therapeutics in clinical use and under clinical investigation.

These are not the final page numbers!

Table 2: Anticancer drugs currently in clinical use.[a]

These are not the final page numbers!

triblock copolymers, which could effectively immobilize

These are not the final page numbers!

These are not the final page numbers!

end, Rhov and co-workers syn-

pH value and thereby cause changes to the

These are not the final page numbers!

Thermoresponsive release is among the most extensively 3.2.4. Photosensitive Release

These are not the final page numbers!

compound,[124] by generating an azobenzene-based molecular

These are not the final page numbers!

4.1. In vitro Delivery hydrophobic or electrostatic interactions. Depending on the

These are not the final page numbers!

These are not the final page numbers!

These are not the final page numbers!

MDR.[157] In their study, DOX was used as a chemo-

These are not the final page numbers!

These are not the final page numbers!

These are not the final page numbers!

These are not the final page numbers!

These are not the final page numbers!

These are not the final page numbers!

through, while both positively and negatively

These are not the final page numbers!

These are not the final page numbers!

These are not the final page numbers!

These are not the final page numbers!

These are not the final page numbers!

5.4. Drug Loading 6.1. Protein–Drug Conjugates

These are not the final page numbers!

although the stability of all of them can be tailored.[271, 273]

These are not the final page numbers!

These are not the final page numbers!

These are not the final page numbers!

to polymer, thus the viscosity of the

Dendrimers are tree-like spheri-

These are not the final page numbers!

include chitosan, hya-

These are not the final page numbers!

with succinic anhydride (SA), they

6.7. Phase-Change Materials

These are not the final page numbers!

microbeads, which were then encapsulated in a 1-

6.8. Inorganic Nanoparticles

Inorganic nanoparticles comprise a very impor-

These are not the final page numbers!

endocytosis.[317] The extremely high specific surface areas of

Figure 21. a) Schematic illustration of conjugation of paclitaxel (PTX)

These are not the final page numbers!

7. Summary and Outlook