886 Current Drug Metabolism, 2010, 11, 886-897

Disposition of Nanoparticle-Based Delivery System via Inner Ear Administration

Gang Chen* , Xiao Zhang, Fan Yang and Lei Mu

Department of Pharmaceutics, School of Pharmacy, Guangdong Pharmaceutical University, Guangzhou, China

Abstract: The inner ear is difficult to access by conventional systemic drug delivery due to formidable physiological and anatomic barri-
ers. There is an increasing interest in the treatment of inner ear disorders by topical application of drugs to the inner ear. One of the
most important issues to overcome before full clinical application is the development of smart delivery systems for drugs to the target
sites and controlled release in the inner ear. This is an area where nanoparticles will play an extremely important role. These submicron
particles have exhibited improved biocompatibility, in vivo stability, target specificity, and cell/tissue uptake and internalization of the
encapsulated therapeutic agents, leading to a decrease in the dose required and a decrease in side effects. This unique combination of
properties makes nanoparticles a novel delivery device, which fulfils the requirements for inner ear application. This review will summa-
rize recent findings and applications of various nanoparticle-based systems like poly (D, L-lactic/glycolic acid) nanoparticles, magnetic
nanoparticles, lipid nanoparticles, liposomes, polymersomes, hydroxyapatite nanoparticles, and silica nanoparticles in the field of inner
ear drug delivery. Moreover, the review will provide an insight into the future strategies of nanoparticle-based cochlear drug delivery. In
conjunction, physiological considerations related to inner ear administration will be highlighted. The routes and applications for local in-
ner-ear drug delivery will also be mentioned. In closing, this review will give an overview of the potential future development in inner
ear administration with nanoparticles.

Keywords: Nanoparticles, cochlea, local drug delivery, intratympanic administration, round window membrane, perilymph.

1. INTRODUCTION drugs needed and hence reduced side effects. NPs have been fre-
The advancement of inner ear medicine will require the devel- quently considered as vehicles for selective delivery chemothera-
opment of a means of nontraumatic and nontoxic delivery of thera- peutic agents to tumor cells while sparing normal-functioning cells,
peutic molecules to the cochlea. However, drug delivery to the requiring much less drug and avoiding the adverse systemic effects
cochlea presents a number of technical challenges, which have typically associated with conventional antineoplastic medicines [9,
hindered the development of therapeutic strategies for the treatment 13]. NPs can also provide sustained therapy, as reported by Guz-
of inner ear disorders. Reasons for the difficulty of drug delivery to man et al. [14], who used local intraluminal infusion of biodegrad-
the cochlea include the limited blood flow to the cochlea [1] and the able polymeric NPs for prolonged drug delivery after balloon an-
existence of the blood-inner ear barrier (BIB), which inhibits the gioplasty. A steady and continuous delivery of drugs was achieved
transport of drugs from the blood to the inner ear and represents a by ocular drug delivery targeting the retina and retinal pigment
fundamental obstacle to systemic application [2-4]. The inner ear epithelium using polylactide NPs [15]. In another application,
tissues are isolated from the surrounding organs by a bony con- Lamprecht et al. [16] utilized biodegradable NPs for targeted drug
struction, which allows the topical introduction of drugs by local delivery in treatment of inflammatory bowel disease.
application. Because of these considerations, local drug application Nanotechnology may also prove useful in inner ear medicine
to the inner ear is becoming of increasing interest in basic research with recent better understanding of the pathologies of noise-, toxin-,
and in clinics as a more effective delivery route compared to sys- inflammatory-, viral- and immune-mediated injury and cell death
temic applications. This mode of delivery bypasses the BIB, allow- processes leading to new treatment approaches [17, 18]. To achieve
ing drugs to reach their intended targets such as the hair cells and an ideal outcome with a minimum amount of adverse effects, NPs
the synaptic regions of hair cells [5]. This approach has been re- have been used for local application of therapeutic biomolecules to
ported for the clinical application of both steroids [6] and gentami- the inner ear for various otologic treatments. This review will dis-
cin [7]. Many other agents are in widespread use in animal research. cuss the NPs as potential drug delivery vehicles to the cochlea.
However, this technique has not been widely used in a clinical set- Much attention will also focus on the in vivo disposition of
ting because controlled and targeted drug delivery systems have not nanoparticle-based delivery systems. Finally, this review will sum-
yet been developed. marize the perspectives for future applications of the systems in
One of the current initiatives in medicine is the exploitation of inner ear drug delivery.
nanosized delivery vehicles, nanoparticles (NPs) to complement
2. PHYSIOLOGICAL CONSIDERATIONS
existing therapeutic strategies [8-10]. NPs, as the very name im-
plies, are particles varying in size from 10 to 1000 nm and depend- 2.1. Anatomy and Physiology of The Inner Ear
ing on the end use, may or may not contain a drug molecule. The The ear is divided into three main parts: the external ear, the
unique size-dependent properties of NPs make these materials supe- middle ear and the inner ear See Fig. (1). The external ear consists
rior for applications in many areas of biology and medicine [11]. of the auricle and the ear canal that ends at the tympanic membrane.
Nanoparticle-based drug delivery may offer increased efficacy and The middle ear consists of the tympanic membrane, the tympanic
reduced drug-associated side effects. The increased efficacy will cavity, the auditory ossicles and the Eustachian tube. The inner ear,
occur in part as a consequence of the ability to target the drug, also called the labyrinth because of its complex structure, consists
within the NPs, to the site within the tissue where the therapeutic of sacs and tubules suspended in cavities of the petrous portion of
effect is required [12]. Entrapment of the drug within the NPs and the temporal bone. These structures contain a fluid called the
controlled release at the required site may result in lower doses of endolymph, while the space between the membranous labyrinth and
the bone is filled with the perilymph. The bony labyrinth consists of
*Address correspondence to this author at the Department of Pharmaceutics, two parts: the vestibule that houses the saccule, the utricle, the
School of Pharmacy, Guangdong Pharmaceutical University, Guangzhou semicircular canals, and the cochlea, a spirally coiled structure. The
510006, P.R. China; Tel: +86-20-3935-2117; Fax: +86-20-3935-2129; sense of balance is located in the vestibule while the sense of hear-
E-mail: [email protected] ing is located in the cochlea.

1389-2002/10 $55.00+.00 © 2010 Bentham Science Publishers Ltd.

Disposition of Nanoparticle-Based Delivery System via Inner Ear Administration Current Drug Metabolism, 2010, Vol. 11, No. 10 887

The cochlea is a two and three quarters coiled cavity in the bone confirmed in several studies based on marked concentration differ-
containing a membranous structure filled with fluid. The cochlear ences in chemical composition, some ototoxic substances and tracer
membranous structure comprises three cavities: the scala vestibuli ions between the perilymph and the blood [2, 3, 28]. It has been
connected to the oval window and the middle ear ossicles, the scala suggested that the BIB is functionally similar to the blood-brain
tympani connected to the round window (RW) at the middle ear, barrier in regulating the transport of various substances from the
and finally the scala media being part of the endolymphatic system. blood to the inner ear tissue and maintaining the microhomeostasis
The scala vestibuli and scala tympani are parts of the perilymphatic and functional integrity of the inner ear [29, 30]. Anatomically, the
system. The scala media contains the sound perceiving organ, the endothelial cells of the inner ear blood capillaries linked to each
organ of Corti, a complex structure containing hair cells receiving other by tight junctions represent the main component of the BIB
the hydromechanical energy and converting it to electrochemical [31]. Tight junctions serve as the structural basis of BIB and extend
signals, supporting cells, a basilar membrane and a tectorial mem- the normal function that limits the passage of certain substances
brane as well as nerve fibres connecting the organ to the nearby into the inner ear. Recent studies have indicated that p-gp plays an
situated spiral ganglion. From the spiral ganglion nerve fibres pro- important role as a drug extrusion pump at the BIB sites [32].
ject to the brain for further processing of the auditory signals. The Therefore, the BIB which inhibits the transport of drugs from the
scala media also contains a highly vascularized structure called stria serum to the inner ear, represents a fundamental obstacle to sys-
vascularis, and it is regarded that the endolymph of the cochlea is temic application. For therapies based on these discoveries, it will
formed in this structure. be necessary to develop safe and reliable methods for the delivery
of drugs into the inner ear, bypassing the BIB.

2.3. Round Window Membrane

The RW, which is located within the RW niche, is probably the
most important interface between the middle ear and the inner ear
that can be used for local pharmacological therapy of inner ear dis-
orders. The RW is the most basal end of the scala tympani, and
allows release of hydraulic pressure of perilymph that is caused by
vibration of the stapes within the oval window. The round window
membrane (RWM) is composed of three basic layers: an outer epi-
thelium towards the middle ear, a middle layer and an inner layer of
epithelium, which borders the scala tympani [33]. The 3-layer
RWM participates in secretion from and absorption into the per-
ilymph. The outer epithelium consists of a single layer of cuboidal
cells with tight junctions, the middle layer is composed of connec-
tive tissue with abundant blood and lymph vessels, and the inner
layer is composed of squamous cells [34, 35]. The RWM of various
species differ in their thickness (70 μm in humans, 40–60 μm in
rhesus monkeys, and 10–14 μm in chinchillas) but share a similar
composition [33]. In a previous study, Saber et al. [36] revealed
that the mean thickness of the RWM in normal guinea pigs was
about 10 μm.
Fig. (1). Structure of the inner ear.
The permeability of the RWM has interested otologic research-
The scala media is filled with the endolymph, about 8 μL in ers for decades. This interest was stimulated by the RWM’s fre-
human, which has an ionic composition similar to the intracellular quent implication in the spread of middle-ear infection to the laby-
environment, high in potassium and low in sodium. The fluid space rinth and by the inherent possibilities of using the RWM as a drug
provides a unique environment for the apical surface of the hair delivery system to the inner ear [35, 37, 38]. The drug deposited
cells. The cells that form the borders of the scala media are all con- locally in the middle ear cavity is transported by pinocytosis
nected by tight junctions. The primary function of the scala media through the outer layer of the RWM, where the micropinocytic
is to provide an electrochemical environment that supports the abil- vesicles containing the drug are transported to the connective tissue
ity of hair cells to transduce mechanical motion into electrical po- of the middle layer, from here reaching the perilymph by lymphatic
tentials. It contains a relatively large (100 mV) positive electrical and blood vessels or crossing the inner layer directly to diffuse into
potential. This unique electrochemical environment is created by the perilymph. Other routes of entry include the oval window annu-
the stria vascularis, which runs along the outside of the scala media lar ligament and the small lacunar mesh in the bone wall surround-
and extracts energy from the blood that is needed to create the elec- ing the inner ear.
trochemical battery of the scala media. Most of the functionally Various substances (drugs) such as antibiotics, local anesthet-
important elements of the cochlea are bathed in perilymph, includ- ics, steroids, toxins, cationic ferritin, horseradish peroxidase, inter-
ing the basolateral surfaces of the sensory cells, the neurons, and feron and 1μm latex sphere can traverse the RWM [33, 38, 39]. The
most of the other specialized structures within the cochlea. The passage of substances through this semipermeable membrane is
perilymph is similar to the cerebrospinal fluid (CSF), with which it affected by the membrane’s thickness, morphologic integrity, the
is in diffusional continuity via the cochlear aqueduct in most ani- presence and duration of inflammation, and the molecular weight,
mals, though protein concentration in perilymph is significantly (5 concentration, configuration, liposolubility, and electrical charge of
to 20 times) higher than in CSF [19-24]. Perilymph volume in the the substances [38, 40]. Because the RWM is relatively well per-
guinea pig is 15 μL [25], while that in the human is 160 μL [26]. meable, the delivery of drugs to inner ear by applying them directly
onto the RWM is a promising way to treat human inner ear disor-
2.2. Blood–Inner Ear Barrier ders [41]. The method is also feasible, as suggested by reports on
In many respects, the inner ear is similar to the brain. It is iso- gene delivery through intact RWMs and expression in the cochlea
lated physically and anatomically from the remainder of the body [42, 43].
by the BIB. The concept of a barrier system in the inner ear was The local application of drugs to the middle ear in a clinical
first proposed by Hawkins [27]. The existence of BIB has been setting, where variations in surgical protocols between institutions,
888 Current Drug Metabolism, 2010, Vol. 11, No. 10 Chen et al.

or in the use of suction in the middle ear, could influence drug lev- CSF after the local delivery of drug NPs or solution were respec-
els achieved in the perilymph. Therefore, it is perhaps not surpris- tively 2.5 and 4.3-fold higher than after systemic application. In
ing that a review of the literature on the RWM permeability uncov- meanwhile, the AUC of interferon in the CSF and brain tissues after
ers data that sometimes seem contradictory and are often difficult to the local delivery were respectively 7.7 and 4.5-fold higher than
interpret. The variations in perilymph concentrations are mainly after systemic application. These results indicate that inner ear ad-
based on a significant variability in RWM permeability. Therefore, ministration may offer a promising alternative to brain-targeted
the quantification of RWM permeability is extremely important as delivery of micro- and macromolecular drugs.
this factor appears to dramatically influence the perilymph levels of However, another studies yield conflicting results. No signifi-
drug achieved. Studies using methods other than fluid aspiration, cant differences were found in the average concentrations of seven
and thereby avoiding or at least minimizing sampling artifacts also selected biochemical substances within the perilymph following
suggest a considerable inter-animal variation in RWM permeability. cochlear aqueduct occlusion [20]. Ohyama et al. [61] suggested
In addition, Nordang et al. [44] concluded in their study that any when the cochlea was in the normal and sealed state, volume flow
local application initiated a certain swelling of the RWM, even through the aqueduct was very low and made little contribution to
saline in the middle ear inducing the RWM thickening [36, 44]. The perilymph homeostasis. Therefore, the question of whether the
implication of swelling to the passage of drugs is not clear. It could cochlear aqueduct provides a physiological biochemical communi-
be speculated that the swelling, despite making the membrane cation between the CSF and the perilymph remains controversial
thicker, actually would make the tissue more loose and permeable and unresolved. Consequently, any study designed to assess phar-
thus facilitating drug entry into the inner ear fluids. macokinetic profiles of chemicals in the inner ear fluids should also
The transfer across the RWM barrier limits the amount of drug include similar profiles of the CSF.
entering the perilymphatic space. Especially for larger molecules or
particles, this has been proven to be of significance as demonstrated 3. LOCAL INNER-EAR DRUG DELIVERY
by the more efficient gene transfer after intracochlear inoculation as Currently, otologic practice relies heavily on both systemic and
compared to RWM application [45]. Uptake of medication through local drug delivery routes. From a clinical perspective, a pharma-
the RWM may be enhanced by the use of certain facilitators [46]. cological protection with systemic administration is preferable.
Among these facilitators, histamine has potential vasodilatory and Systemic steroids, for example, is used in the otologic management
increased permeability effects on the RWM, hyaluronic acid has of idiopathic sudden and immune-mediated sensorineural hearing
known osmotic effects, and dimethylsulfoxide, an organic solvent, loss (SNHL), which has been proved with success and is one of the
may increase solubility of medication in perilymph [47-49]. current standard treatment options. Their clinical usefulness, how-
ever, is limited by undesirable side effects arising from the high
2.4. Communication Between the Perilymph and the Cerebro- systemic doses required to achieve sufficient cochlear fluid levels
spinal Fluid of drug to produce the intended inner ear effects [62]. The possibil-
Perilymph seems to have three different potential origins, alone ity to pharmacologically prevent, or treat inner ear disorders is sig-
or in combination [19, 50, 51]. One source is the CSF that reaches nificantly restricted due to the presence of certain physiological and
the cochlea via the cochlear aqueduct. The cochlear aqueduct con- anatomical barriers such as limited cochlear blood supply and the
nects the perilymphatic compartment with the CSF of the cranium presence of the BIB. Thus, if administered systemically, the avail-
and opens into the basal turn of scala tympani. CSF may reach and ability of the drug for the target cells within the cochlea may be
mix with perilymph of the scala tympani, though the transfer of compromised. In addition, systemic drug administration is associ-
fluid between the two spaces may be limited [52]. Sterkers et al. ated with increased risk of systemic side effects. Other disadvan-
[53] suggested that the origin of scala tympani perilymph was dif- tages of a systemic therapy are (1) many drugs have short plasma
ferent from that of scala vestibuli perilymph. They noted an altera- half-lives, (2) if their molecular weight is high, the parenteral
tion in the composition of the scala tympani perilymph after block- resorption is impaired, and (3) there is large inter-individual varia-
ing the cochlear aqueduct. In some lower-order mammals, the aq- tion due to variation in dosage and body metabolism.
ueduct maintains its relative patency. In humans, the aqueduct is In some ways, the inner ear is particularly well suited for local
usually not patent [54], though diffusional continuity is achieved drug delivery. It is isolated from the remainder of the body by the
via other pathways, most notably the spaces around and within the BIB, and the perilymph and endolymph fluids permit liquids to
auditory nerve [55]. The second source of perilymph is the CSF that reach the entire cochlea quickly. The local delivery into the inner
enters the cochlea through perivascular spaces and vestibulo- ear has been the basis for a large number of advances in under-
cochlear nerve sheaths at the distal end of the internal auditory ca- standing cochlear function and provides a potential avenue for
nal. The third and probably the major source comes from the blood therapeutic drug. This mode of delivery bypasses the BIB, allowing
vessels that supply the inner ear itself. drugs to reach their intended targets more directly with lower doses.
Simulations demonstrated that the substance with RWM injec- Thus, a higher concentration of the drug in the inner ear can be
tion could transport into the CSF due to the injection site being obtained, and systemic side effects of the drug are eventually pre-
closer to the cochlear aqueduct, hence the lower levels of the in- vented. At present, there are two main routes for local drug delivery
jected substance in the perilymph [56]. Indeed, it has been sug- to the inner ear: (1) intracochlear or intravestibular drug delivery,
gested that higher rates of inner ear infusion are not only ineffective and (2) intratympanic (extracochlear) drug application to the RWM.
in improving drug distribution within the cochlear partition; they Because opening of the human inner ear for the sole purpose of
result in displacement of the infusion solution through the cochlear local drug delivery is rarely performed, intracochlear or intravesti-
aqueduct and into the CSF [57]. Spread of NPs to the CSF does bular drug delivery is generally limited to situations where the inner
occur as demonstrated by the transmission of NPs to the contralat- ear is already opened for another reason, such as cochlear implanta-
eral cochlea. This has been described in gene vector systems [58]. It tion [63], or where the applied drug is intended to be ablative [64].
implies another potential danger of one perilymphatic injection To overcome these problems, an alternative approach is to inject the
technique is that the injected materials could enter the cochlear drug directly into the inner ear fluids. The present study is focused
aqueduct and travel through the CSF to the contralateral ear where on developing methods for injecting drugs directly into the cochlea
they could cause unintended effects. Chen et al. [59, 60] investi- with minimal trauma and leakage. Intratympanic management,
gated the feasibility of delivering the drugs (dexamethasone and introduced over 50 years ago [64], has frequently been used for
interferon) to the brain (CSF) via the inner ear. Their studies re- the therapy of the various inner ear diseases. In this method, it is
vealed that the area under curve (AUC) of dexamethasone in the
Disposition of Nanoparticle-Based Delivery System via Inner Ear Administration Current Drug Metabolism, 2010, Vol. 11, No. 10 889

regarded that the given drug shows its effect passing through enabling the drug direct delivery to the desired cell population. The
the RWM into the fluid spaces of the inner ear. small size and inaccessibility of the cochlea impose constraints on
Applications for inner ear drug delivery are focused on four the development of a device to deliver drugs to the inner ear. Possi-
main fields: otoprotection, hearing loss, autoimmune inner ear dis- ble way of reaching this goal is the use of nanoparticle-based deliv-
ease, and regeneration of hair cells. Recent advances in the pharma- ery systems to regulate the drug release and manipulate the target
cology and molecular biology of the inner ear have revealed new sites within the inner ear. Encapsulation of therapeutic agents into
possibilities for preventing or treatinging inner ear disorders. Scien- NPs would provide controlled release. NPs applied locally to the
tists and clinicians are making progress in understanding the mo- inner ear would diffuse in the inner ear fluids without being cleared
lecular mechanisms associated with cochlear and auditory nerve from them. The delivery carriers would release loaded drugs with
degenerative processes [65] and in finding agents that can minimize either through biodegradation or diffusion out of the carriers. Other
degeneration and facilitate repair [17, 18, 66, 67]. The extraordinary solutions to drug targeting could include NPs functionalized with a
progress that has been made in defining the genes and protein ac- variety of ligands to direct a drug to a specific cell type or using
tivities offers hope for gene transfer [68] and molecular approaches magnetic NPs, the dispersal of which can be influenced by external
[69] to treat these diseases. Therefore, more candidate substances magnetic fields. The main nanoparticle-based systems in inner ear
for the treatment of inner ear disorders are being discovered. Table drug delivery are presented in Table 2.
1 summarizes the main medications applied in inner ear delivery.
4.1. Poly (D, L-Lactic/Glycolic Acid) Nanoparticles
There is large variability in the reported results of local inner-
Polymeric NPs are included in the first generation novel colloi-
ear drug delivery trials [97]. It may result from factors such as mid-
dal carriers developed with the objective to improve drug delivery.
dle ear volume, clearance of the drug from the middle ear cavity via
Poly (D,L-lactic/glycolic acid) (PLGA) polymers are attractive as
the Eustachian tube, and condition of the RWM as well as the per-
carriers, because of their hydrophilicity, biocompatibility, promo-
meability and contact of the drug with the RWM [98]. Specifically,
tion of cell membrane endocytosis and relative ease of derivitiza-
the presence of middle ear clearance will affect the drug levels
tion with functional groups attached on the inside or outside of the
achieved in the perilymph for all single or repeated intratympanic
polymer. Programmed degradation of the polymer could result in
injections of defined drug volumes. When drug fluid remains in
timely, quantitative delivery of a drug, plasmid, or other bioactive
contact with the RWM, this allows the drug to diffuse into or out of
molecule.
the perilymph according to the prevailing concentration gradient. If
drug fluid is removed from the RW niche, only a small concentra- Given these findings, Tamura et al. [100] examined the poten-
tion peak is reached very early at the very basal part of the scala tial of PLGA NPs for use as a vehicle for delivering drugs to the
tympani. In order to establish the perilymph clearance rate defini- cochlea. They prepared PLGA NPs encapsulating rhodamine, a red
tively, additional data are necessary in which a sustained drug level fluorescent dye, and administered these systemically or locally to
is maintained on the RWM so that perilymph kinetics are not influ- adult guinea pigs. The systemic application of NPs had a significant
enced by middle ear clearance. Efforts to control this variability and effect on targeted and sustained delivery of rhodamine to the liver
to overcome the limitations associated with local drug delivery to but not the kidney or cochlea. Rhodamine NPs placed on the RWM
the inner ear, have led to the development of a variety of delivery were identified in the scala tympani as NPs, indicating that the
vehicles [99]. PLGA NPs can permeate through the RWM. Previous studies have
demonstrated that cochlear fluids have an extremely slow flow rate
4. NANOPARTICLE-BASED DELIVERY SYSTEMS VIA [61]. Rhodamine particles observed in the cochlea after local appli-
INNER EAR ADMINISTRATION cation were located in the perilymphatic space, which indicated that
In local drug delivery to the inner ear, both the drug delivery the clearance of rhodamine particles depended on the flow of per-
system and the application protocol significantly influence the dis- ilymph. If the flow of perilymph was slow, it would follow that the
tribution of substance within the inner ear. Direct topical admini- clearance of rhodamine particles from the cochlea might also be
stration also result in otopathology and the concentration of slow, thereby resulting in sustained release of rhodamine from rho-
drug reaching the inner ear is difficult to control. The goal is to damine particles in the perilymph. The numbers of rhodamine par-
control drug distribution within the cochlea after the local delivery, ticles in the cochlea after local application were apparently higher

Table 1. Medications Applied in Inner Ear Delivery

Medications Indications Results Refs.

Hearing loss, Meniere’s Steroids accumulated in the inner ear in much larger concentrations and remained longer after [6, 46, 62,
Steroids
disease and tinnitus local delivery than after oral or intravenous administration. 70-77]
Reactive oxygen species
Antioxidants delivered systemically with the cisplatin resulted in hearing protection but also
and oxidative damage,
made the cisplatin ineffective. However, local administration to the inner ear of antioxidants
Antioxidants such as ototoxicity and [78-81]
could protect hair cells without inactivating the chemotherapy treatment and give high local
acute noise-induced
concentrations of the substances in the perilymph.
hearing loss
Neural stem cells survived in normal rat cochlea after transplantation via round window mem-
Neural stem cells Lost hair cells [82-84]
brane and showed no obvious effects on auditory functions of the rat cochlea.
Hair cells and spiral gan-
Neurotrophins have a short serum half-life and have difficulty reaching inner ear tissues. How-
Neurotrophic factors glion neurons degenera- [67, 85-88]
ever, local inner-ear delivery could introduce them directly into the cochlear fluid.
tion
In vivo drug delivery to introduce exogenous DNA or RNA into the inner ear was achieved using
Deafness and genetic local approaches. Several vectors were used to deliver genes into the inner ear, including cationic
Genes predisposition for postna- liposome vector, adenovirus vector, adeno-associated virus vector, herpes-simplex virus vector, [89-96]
tal loss of hearing and vaccinia virus vector. Safe and effective vectors of local gene delivery to the cochlea could
facilitate the clinical application of these therapeutic treatments.
890 Current Drug Metabolism, 2010, Vol. 11, No. 10 Chen et al.

Table 2. Different Nanoparticle-Based Systems in Inner Ear Drug Delivery

Nanoparticle based
Species Comments Refs.
System

PLGA NPs accumulated in the cochlea after local application in much larger numbers than after
systemic application and slowly released rhodamine. Therefore, the local application of PLGA
PLGA NPs Guinea pigs [100]
NPs to the RWM may be an effective strategy for targeted and sustained drug delivery to the
cochlea.
Guinea pigs/ Magnetic NPs can reach the cochlea through the RWM under an external magnetic field, which
Magnetic NPs [101-108]
Wister rats/ chinchilla provide an entry for delivery of bioactive molecules to target specific inner ear tissues.

The lipid NPs may permeate the RWM and reach the spiral ganglion cells, nerve fibers, hair cells
Sprague-Dawley rats/
Lipid NPs and spiral ligament, which indicate great potential of NPs towards significant sustained-release [59, 109-111]
guinea pigs
and targeting the inner ear and central nervous system via inner ear administration.
The liposomes for cochlear gene transfer provide a safe and rapid alternative to the use of recom-
Liposomes Mice/ guinea pigs binant viral vectors in gene therapy for inner ear disorders. Compared with viral vectors, how- [43, 89-91]
ever, liposome-mediated gene delivery has been inefficient.
The multifunctional polymersomes could be a potential tool for cell specific drug and gene deliv-
Polymersomes Mice ery to the inner ear. Further work need be done to incorporate therapeutic agents into the func- [112]
tionalized polymersomes.
The hydroxyapatite NPs can mediate gene transfection both in vitro and in the cochlea of the
Hydroxyapatite NPs Guinea pigs/mice [113-115]
living animal. However, its transfection efficiency needs to be improved.
The Cy3-labeled silica NPs were taken up in inner hair cells, vestibular hair cells, spiral ganglion
neurons, and supporting cells after RWM delivery. Due to their different characteristics, how-
Silica NPs Mice [116]
ever, DNA-silica NPs complexes may show a different distribution pattern or even fail to trans-
fect inner ear structures.

PLGA: Poly (D, L-lactic/glycolic acid), NPs: nanoparticles, RWM: round window membrane

than after systemic application. In addition, Rhodamine was also model that with magnetic NPs indomethacin had 60-fold higher
found from the basal to apical portion of the cochlea after a local concentrations and considerably reduced drug concentration in non-
injection of rhodamine NPs into the scala tympani, whereas it was target organs [119]. Furthermore, magnetically susceptible NPs,
observed only to a limited degree in the basal portion of the cochlea controlled by an external magnetic field, have the ability to reach
after direct application of rhodamine NPs to the RWM. The rho- target tissues that are difficult to access, such as the inner ear.
damine released from PLGA NPs in the perilymphatic space possi- Mondalek et al. [101] designed an in vitro RWM model similar
bly spread more toward the apical portions of the cochlea than to the RWM to detect the feasibility of using SPION for targeted
PLGA NPs applied to the RWM. delivery of therapeutics to the inner ear. The RWM model was a 3
Therefore, the local application of PLGA NPs to the RWM may cell-layer model with epithelial cells cultured on both sides of a
be an effective strategy for targeted and sustained drug delivery to small intestinal submucosal matrix and fibroblasts seeded in be-
the cochlea. Therapeutic molecules for inner ear diseases can be tween. Dextran encapsulated NPs clusters 130 nm in diameter were
encapsulated in PLGA NPs, which applied as local drug delivery pulled through the RWM model using permanent magnets. The
systems. Further studies are required to optimize the profile of NPs results demonstrated that SPION had been magnetically transported
in accordance with the desired distribution and release of drugs they through this model, allowing quantitive evaluation of prospective
contain. targeted drug or gene delivery through the RWM. In addition,
SPION showed no toxicity to cells in culture. To quantitatively
4.2. Magnetic Nanoparticles compare in-vitro and in vivo membrane transport studies of targeted
In addition to the use of magnetic NPs as MRI (magnetic reso- delivery, it needed characterization of the magnetically induced
nance imaging) contrast agents, NPs can be used for magnet guided mobility of SPION [102]. In a study, flux densities, gradients, and
targeting of the NPs themselves or of cells containing these NPs for NPs properties were measured in order to quantify the magnetic
a variety of applications [10, 117]. Since the first magnetic polym- force on the SPION in both an artificial cochlear RWM model and
eric particles of the 1970s, numerous NPs have been developed to the guinea pig RWM. The force (9.69
10-20 N) required to achieve
deliver therapeutic substances to localized disease sites. The use of in vivo transport was significantly lower than that (5.04
10-16 N)
superparamagnetic iron oxide nanoparticles (SPION) for the deliv- required to pull NPs through the in-vitro RWM model. Indeed very
ery of therapeutic molecules holds potential clinical applications. little force was required to accomplish measurable delivery of po-
Synthetic, -Fe2O3 (maghemite) or Fe3O4 (magnetite) particles with lymeric-SPION composite NPs across the RWM. SPION hierarchi-
a core less than 15 nm in diameter will exhibit superparamagnetism. cally coated with oleic acid and Pluronic F127 copolymers (POA:
SPION have no residual magnetic interaction in the absence of a PION) have shown exceptional T2 contrast enhancement [103].
magnetic field (unlike larger ferromagnetic particles) and have POA: PION was administered through intracochlear, intratympanic
multiple applications in biomedicine [118]. Molecular-sized SPION and intravenous routes in male Wister rats. POA:PION was intro-
can locate in proximity to a biological target, be derivitized to pro- duced into the perilymph space, after which it became widely dis-
mote biocompatibility or interaction with target cells, and are sus- tributed and demonstrated the integrity of the perilymph-endolymph
ceptible to an external magnetic field gradient. For in vivo applica- barrier. POA:PION passed through the middle-inner ear barriers in
tions, organic coatings surrounding SPION are used to insure dis- only small amounts, but stayed in the perilymph for 3 days. They
persion and prevent aggregation of particles, which would impede did not traverse the blood-perilymph barrier or blood-endolymph
transport across cell membranes. Today, SPION are synthesized, barrier. The initial results by Kopke et al. [104] with in vitro model-
then encapsulated in either silica or PLGA or obtained commer- ing, organotypic culture modeling, and in vivo studies suggested the
cially with coatings of oleic acid or dextran. Others found in a rat following: (1) SPION could be pulled through a tripartite in vitro
Disposition of Nanoparticle-Based Delivery System via Inner Ear Administration Current Drug Metabolism, 2010, Vol. 11, No. 10 891

membrane model, living guinea pig and rat RWM, and fresh ca- supporting cells. Future successes of SPION applications in
daver RWM in less than 60 min by a magnetic force of ～0.3 tesla; nanomedicine, like other biomaterials, will be related to the extent
(2) magnetic gradient-forced transport superseded diffusion or ac- of complete physicochemical characterizations (e.g. zeta potential)
tive forces of RWM transport; (3) SPION and PLGA particles ap- since surface chemistry dictates cell differentiation.
peared biocompatible to inner ear tissues and to the RWM; (4) a
payload had also been transported across the RWM in vivo. 4.3. Lipid Nanoparticles
Future studies were aimed at gathering additional quantitative Despite about 35 years of intensive research, the polymeric NPs
information. Hence, Magnetic NPs were conjugated with 99mTc never made it to the pharmaceutical market. Possible reasons in-
and solution placed on the RWM of anesthetized guinea pigs [105]. clude difficulties in scale-up, high cost of biodegradable polymers,
The RWM was parallel to the pole face of an external magnet. Con- potentially toxic/allergic end products of biodegradable polymers.
trol animals were tested with the same solution, without magnets. The lipid NPs do not have these problems [120]. They are interest-
Significantly, more NPs were pulled into the cochlea by magnetic ing delivery systems, which possess a lipid matrix with a controlled
targeting in this preliminary study, 330% above controls. Calcula- structure for optimising drug incorporation and modifying drug
tions of quantitive delivery were performed such that if individual release.
therapeutic molecule loading was known, delivery of drugs could Zou et al. [109] evaluated the ability of the lipid nanocapsules
be calculated. Ge et al. [106] investigated the inner ear distribution to pass through the RWM and reach inner ear targets after Sprague-
of PLGA-encapsulated SPION in the chinchilla cochlea after its Dawley rat RWM administration. The results demonstrated that the
application on the chinchilla RWM. PLGA-SPION passed through lipid nanocapsules could permeate the RWM and reach the spiral
the RWM and appeared in the perilymph at the scala tympani with ganglion cells, nerve fibers, and spiral ligament fibrocytes within 30
or without exposure to magnet forces. These particles might have min. The main pathway for the lipid nanocapsules to reach hair
then translocated to the upper cochlear turns by diffusion, as was cells was the ‘‘nerve pathway,’’ that was, diffusion from perilymph
evidenced by the finding of particles in the apical cochlear turn. in the scala tympani to the spiral ganglion, continuing with nerve
Further, particles crossed the boundary of cochlear duct and entered fibers inside Rosenthal’s canal to approach the inner hair cells and
the organ of Corti, as evidenced by their appearance in the basilar pillar cells, and finally the outer hair cells. The rapid accumulation
membrane, Reissner’s membrane, and the endolymph. Finally, NPs of the lipid nanocapsules in the spiral ganglion cells indicated that
penetrated the hair cells and supporting cells through the apical or the most likely pathway was directly through the porous modiolar
basolateral membranes, as evidenced by their presence at cuticular wall of the scala tympani [55]. The uptake of the lipid nanocapsules
plate of an outer hair cell and at the base of an inner hair cell. NPs in the tunnel fibers originated from either the spiral ganglion or the
were also seen in the lateral wall structures. The perilymphatic and basilar membrane. The lipid nanocapsules on the surface of nerve
endolymphatic fluids could be the source for NPs entering to spiral fibers also freely diffused between different fibers including affer-
ligament and stria vascularis, respectively with or without magnetic ent and efferent nerve fibers and migrated along the longitude of the
force. Silica-encapsulated magnetite NPs were synthesized, charac- nerve fibers. The disappearance of the lipid nanocapsules from the
terized and tested for biocompatibility in a non-dividing cell model tunnel fibers after long time incubation with solutions indicated
and in organotypic (three day mouse pup) Organ of Corti (OC) their surface location. The delayed distribution of the lipid nano-
cultures [107]. The MATH-1 gene was inserted into a plasmid with capsules in the hair cells came from the spiral ganglion, the tunnel
promoters to facilitate intracochlear transfection and this plasmid fibers, and the Corti’s liquid. The gradient uptake of the lipid nano-
was tested for viability in a non-dividing cell line. The plasmid was capsules from the inner hair cells to the outer hair cells (i.e., quicker
combined with NPs in a carrier for RWM transport. The NPs and greater uptake in the inner hair cells than that in the outer hair
moved into the target cell using an external magnetic field that was cells) suggested that the lipid nanocapsules chiefly came from the
controllable to move the NPs in three dimensions through the body spiral ganglion area via Rosenthal’s canal because of more intense
and the bioactive substance was released from the NPs once inside innervation and a shorter distance to the inner hair cells. The lipid
the target cell. It was feasible for using external magnetic fields to nanocapsules also traveled through the retrograde pathway from the
pull superparamagnetic NPs with an associated gene into the per- stria vascularis to Corti’s liquid because there was a rapid, intense
ilymph [108]. uptake of the lipid nanocapsules in the stria vascularis. The
These results showed the pathway of magnetic NPs transport ‘‘paracellular pathway’’ was the main approach for the lipid nano-
through the RWM and inner ear structures was not completely capsules penetration of the RWM. The lipid nanocapsules also
clear. These particles might readily pass through intracellular and reached the vestibule, middle ear mucosa, and the adjacent artery.
intercellular pathways simply by diffusion across concentration The vestibular distribution of the lipid nanocapsules originated
gradients, especially for the smaller particle sizes. NPs penetrate the from the spiral ligament. The uptake of the lipid nanocapsules in
hair cells and supporting cells in the organ of Corti through the the spiral ligament fibrocyte type V, which is in contact with the
intracellular pathway of these cells. Clusters of aggregated NPs perilymph in the scala vestibule, provides evidence for this pathway
might collect as string aggregates in the inner ear after passing the [121, 122].
RWM. Individual magnetic NPs become magnetized as induced in These results suggest that the lipid nanocapsules are potential
magnetic fields and tend to agglomerate but do not remain magnet- vectors for drug delivery into the spiral ganglion cells, nerve fibers,
ized when the field is removed. Pinocytosis, however, might be the hair cells, and spiral ligament. Thus, there is a possibility that drugs
mechanism for the larger clusters of NPs that are observed in vari- can also reach the vestibule by applying the lipid nanocapsules to
ous locations. the RWM. The distribution of the lipid nanocapsules in the middle
Magnetic targeting of NPs through the cochlear RWM may ear mucosa and stapes artery wall compromises the relative cell
provide an entry for delivery of bioactive molecules. It may be that population selectivity of the lipid nanocapsules. This can be further
compounds or drugs that possess therapeutic properties can be en- overcome by limiting the lipid nanocapsules in the area of the RW.
capsulated or conjugated to NPs, either to target specific inner ear It is easier to achieve in humans because the RW area is much
tissues or to elicit widespread effects throughout the inner ear. larger than that in rats.
Though these benefits are attractive, little progress has been made Scheper et al. [110] examined the lipid nanocapsules and hy-
towards the goal of using SPION as in-vivo carriers of therapeutic perbranched polylysine for their cellular uptake and possible toxic-
payloads. The next step will involve loading the SPION with a ity in vitro and in vivo. The study showed both NPs were detectable
therapeutic biomolecule, looking for timed, quantifiable, targeted in the fibroblasts' cytoplasm without causing cytotoxic effects. Af-
release of the biomolecule into the perilymph for access to inner ear ter intracochlear application, they were visualized in cochlear cells,
892 Current Drug Metabolism, 2010, Vol. 11, No. 10 Chen et al.

which did not lead to a change in hearing threshold or loss of hair plasm and translocation to the nucleus are considered to hinder
cells. Biocompatibility and traceability were demonstrated for the expression of liposome complexed genes. Expression of most
lipid nanocapsules and hyperbranched polylysine. Thus, the lipid transgenes delivered by the liposomes has been shown to last for a
nanocapsules comply with the basic requirements for drug carriers week or less. However, the shortcomings associated with the use of
for potential application in the inner ear. the liposomes are significantly counterbalanced by a number of
In the studies by Chen et al. [59, 111], dexamethasone acetate- critical attributes. The liposomes are non-immunogenic, easily pre-
loaded solid lipid nanoparticles (SLN) were administered by intra- pared in large amounts and can then be mixed with DNA of virtu-
tympanic and intravenous routes in guinea pigs. After intravenous ally any size to yield complexes that are held together through ionic
injection of SLN, dexamethasone failed to be detected in the per- interactions. The lipoplex does not replicate or recombine and in
ilymph. Compared with dexamethasone sodium phosphate-loaded general does not integrate into the genome of the host cell. Hence,
in situ gel following intratympanic injection [123], the relative the liposome bound DNA poses minimal risk of insertional
bioavailability of dexamethasone in the perilymph was 504% fol- mutagenesis. In addition, the use of liposomes may be limited be-
lowing intratympanic injection of SLN, which also resulted in in- cause of problems related to stability, the inability to deliver to the
creased the t1/2 and the mean residence time by 0.5 and 1.9 folds. right site, and the inability to release the drug when it gets to the
Moreover, SLN increased the AUC by 13 times and extended the right site. However, liposome surfaces can be readily modified by
mean residence time by 19 times in the CSF than the drug solution attaching PEG units to the bilayer (producing what is known as
following intratympanic administration. All these studies indicate stealth liposomes) to enhance their circulation time in the blood-
great potential of SLN towards significant sustained-release and stream. Furthermore, liposomes can be conjugated to antibodies or
targeting the inner ear and central nervous system via inner ear ligands to enhance target-specific drug therapy.
administration.
4.5. Polymersomes
4.4. Liposomes Amphiphilic block copolymers, consisting of chains of hydro-
Lipid bilayers of liposomes are similar in structure to those phobic and hydrophilic units, self-assemble into polymersome NPs
found in living cell membranes. Liposomes offered several advan- in an aqueous environment [124]. Structurally polymersomes are
tages such as encapsulation of hydrophobic and hydrophilic drugs, similar to liposomes. However, they have several advantages over
controlled drug release and reduction in toxicity/increased therapeu- liposomes, which give them greater potential as drug delivery vehi-
tic efficacy of drugs most of which were not offered by submicronic cles. The membrane thickness can be controlled by the molecular
emulsions [120]. weight of the hydrophobic block copolymer, to achieve thicker,
stronger membranes making them more stable than conventional
In the inner ear, there is not much information dealing with liposomes. The polymersome surface can be functionalized with
liposomes application. However, a few studies have shown that the targeting moieties. The main purpose of functionalization of the
liposomes (cationic liposomes) can be further developed in non- hydrophilic corona is to modulate biodistribution of polymersome
viral vector gene therapy. An approach used has been to transfect a NPs and induce cellular uptake by receptor-mediated endocytosis.
liposome mixture with LacZ or green fluorescent protein (GFP) Targeted uptake of multifunctional NPs in a cell, tissue or disease
reporter genes into mice and guinea pig cochleae in vivo [89-91]. specific manner represents a potentially powerful technology in
Wareing et al. [89] studied the feasibility of cationic liposome- biomedicine [8].
mediated gene transfer within the guinea pig cochlea in vivo
following direct microinjection into the cochlea. Transgene The neurotrophin family signals through two distinct groups of
expression was persistent up to 14 days in the neurosensory cell surface receptors the Trks (Tyrosine kinase receptors) and p75
epithelia and surrounding tissue without toxicity and inflamma- neurotrophin receptor [125]. Neurotrophins and their receptors are
tion in the target organ. This study represents the first success- reported to regulate the survival of vestibular and cochlear neurons
ful use of cationic liposomes for cochlear gene transfer thus pro- [126]. Targeting ligands such as short peptide sequences, derived
viding a safe and rapid alternative to the use of recombinant from human nerve growth factor beta, can be conjugated to the end
viral vectors in gene therapy for inner ear disorders. Maeda et al. of the hydrophilic segment so that they may extend outward from
[43] used a liposome-complexed plasmid vector that express the NPs corona. The targeting ligands then readily encounter and
GJB2R75W was applied onto the intact RWM, in order to vali- interact with membrane surface receptors Trks and p75 neurotro-
date the transgene delivery and expression through the RWM phin receptor. Roy et al. [112] investigated the utility of a Nerve
and to evaluate the feasibility of the method for testing hearing Growth Factor-derived peptide (hNgf EE) functionalized polymer-
phenotype. Access of the liposomal complexes used to scala somes to target cells of the inner ear. These functionalized polymer-
media from the scala tympani was possible through the ca- somes were introduced to organotypic explant cultures of the mouse
naliculi perforantes of the osseous spiral lamina, which pro- inner ear and to PC-12 rat pheochromocytoma cells. The polymer-
vided access to the basal domain of supporting cells in the or- somes did not show any obvious signs of toxicity in both organo-
gan of Corti. GJB2R75W-eGFP transgene expression was ob- typic culture and PC-12 cells. Specific targeting and higher binding
served in the endothelial cells in the basilar membrane, inner affinity to spiral ganglion neurons, Schwann cells and nerve fibers
and outer pillar cells, outer hair cells, Claudius cells, spiral lim- of the explant cultures were achieved through ligand mediated mul-
bus and ligament, while expression was equivocal in the Reiss- tivalent binding to Trks and p75 neurotrophin receptors. Unspecific
ner’s membrane and the spiral ganglion. Jero et al. [91] also uptake of polymersomes was investigated using NPs conjugated
found that a liposome-hrGFP could pass the RWM into the inner with scrambled hNgf EE. There would be two possibilities for a
ear and transgene was detected in Reissner’s membrane, spiral lim- successful release of the cargo from the polymersomes: (1) po-
bus, spiral ligament, and spiral ganglion cells without damage to the lymersomes might remain within the endosome but the drug found
middle ear and inner ear in the mouse. This methodology may the way into cytoplasm [127]; (2) polymersomes might escape from
ultimately have applicability to human deafness. Transgene ap- the endosome and release the drug into the cytosol. It appeared that
plication on the intact RWM is appealing because the RWM is polymersomes with scrambled hNgf EE could not escape from the
easily accessible and this delivery route does not require open- endosomal vesicles, hence showed spotty distribution. As hNgf EE
ing of the inner ear, thereby reducing the possibility of iatro- had the specific targeting ability to the Trks and p75 receptors, it
genic damage. could be contemplated that hNgf EE conjugated polymersomes
were uptaken by the receptor-mediated pathway, which might have
Compared with viral vectors, however, liposome-mediated gene an influence in the endosomal escape. These results indicate a se-
delivery has been inefficient [90]. Inefficient DNA entry into cyto- lective cochlear cell targeting by multifunctional polymersomes,
Disposition of Nanoparticle-Based Delivery System via Inner Ear Administration Current Drug Metabolism, 2010, Vol. 11, No. 10 893

which can be a potential tool for cell specific drug and gene deliv- inner ear structures due to different characteristics of Cy3-labeled
ery to the inner ear. Further work need be done to incorporate silica NPs and DNA-silica NPs complexes. The expression of genes
therapeutic agents into the hNgf EE functionalized polymersomes via NPs delivery will be much more complex. These complexes
and the biological effect will be studied. will have to be studied before expanding the use of silica NPs in the
inner ear.
4.6. Hydroxyapatite Nanoparticles
Hydroxyapatite [HAp; Ca10(PO4)6-(OH)2] NPs which have 4.8. Future Strategies
been used for various biomedical applications, are chemically simi- While a delivery using a carrier can prolong the stay of the drug
lar to the mineral component of bones and hard tissues in mammals. within the middle ear cavity, it does not ascertain control of direct
HAp-NPs may promote the adhesion, cellular function, osteocon- contact between the drug vehicle and the RWM. An important vari-
ductivity, proliferation and synthesis of alkaline phosphatase in able is the proximity and constancy of contact between the drug and
osteoblasts and lead to more rapid repair of hard tissue injury [128- the RWM. Thus, the duration of contact with the RWM is a major
130]. problem in case of transtympanic administration [98]. NPs may be
Zhu et al. attempted to use HAp-NPs as gene carriers [131]. cleared from the middle ear cavity via the Eustachian tube, thereby
The results showed NPs-DNA complex could transfer EGFP-N1 diminishing the efficacy of the delivery system.
into the SGC-7901 cells, and transport into the liver, kidney, and Hydrogel carrier with its biological properties can promote
brain after intravenous administration. Sun and Jiang first used increased contact and duration of exposure with the RWM and
HAp-NPs as a novel vector for inner ear gene therapy [113, 114]. eliminate thus some of these variables. A better and safer way
HAp-NPs could load recombinant plasmid enhanced green fluores- would be to disperse the NPs in a hydrogel of a mucoadhesive/
cent protein C2-neurotrophic factor-3 (pEGFPC2-NT3), and protect biodegradable polymer like chitosan or sodium hyaluronate for
the genes from being destroyed by DNase I. Methylthiazolyl tetra- placing in the tympanic cavity. The polymers can be used to encap-
zolium assay confirmed well biocompatibility between HAp-NPs sulate the NPs, which have the ability to provide controlled release.
and the cultured Hela cells. Immunoflourescence test of the report NPs entangled in polymeric matrix would remain unaffected while
gene GFP revealed that the transfection efficiency of HAp-NPs the hydrogel degrades. As the process of biodegradation/dissolution
vector in Hela cells was 15.7%
2.6%. EGFP was immunofloures- of the polymer proceeds, the NPs would release their drug content
cently expressed by transfection of HAp-pEGFPC2-NT3 in the into the matrix, thus providing a continuous release of the loaded
primarily cultured cochlear neurons of mice. HAp-NPs consisting drug. It may be useful as biodegradable tympanic implants to take
of pEGFPC2-NT3 were perfused perilymphatically into the cochlea advantage of long lasting release.
of the guinea pigs with experimental excitotoxic inner ear damage. In a different system, a modified NPs-in-situ-gel can also be
HAp-pEGFPC2-NT3 complex transfected into spinal ganglion cells envisaged. Application of in situ gel has been established in tym-
of the cochlea. In the mean time, the auditory brain-stem response panic drug delivery [123]. Fabricated NPs can be suspended in an
was partly restored compared with that after the injury of the exci- in situ gelling vehicle, which has been characterized for in situ
totoxicity. properties. The NPs dispersed in such a vehicle can be used as to
Compared to the drilling method to the scala tympani, the per- form a gel within the middle ear cavity after intratympanic admini-
meation of drugs through the intact RWM is a safer method for stration. It must be said that at the moment these are concepts, but
clinical therapy. Peng et al. [115] explored the feasibility of HAp- possess high potential for future successful development.
NPs carried NT-3 gene transferring into inner ear through the
RWM and facilitating agents in order to improve the permeability CONCLUSION AND PERSPECTIVE
of RWM. The results demonstrated HAp-NPs carried NT-3 gene Research progress within the last decade is leading to new po-
could pass through the intact RWM, and using hyaluronic acid as tential therapies for a broad spectrum of inner ear disorders. Thera-
facilitating agent could effectively increase the efficiency of trans- pies based on advances in cell and molecular biology will likely
fection of foreign genes into the cochlea. HAp-NPs can mediate revolutionize the treatment. Such treatments may require sequenced
NT3 gene transfection both in vitro and in the cochlea of the living delivery of multiple and potentially unstable therapy agents for
animal. As a non-viral gene vector without the risk of biological periods of months to years. It is likely that many of the newer
disaster, HAp-NPs vector is worthy of further fully researches al- treatments will have broad systemic effects if delivered by conven-
though its transfection efficiency needs to be improved. tional techniques and may be highly specific drugs, such as those
needed to direct cochlear cell repair. The development of local de-
4.7. Silica Nanoparticles livery method offers the possibility that we may be able to deliver
Silica NPs have been found to bind and protect plasmid DNA directly to the inner ear preventive agents, rescue agents, and pro-
[132] and have already been used to transfect Cos-1 cells [133]. No moters of repair and regeneration. Thus, the route of administration
significant cell toxicity has been reported for silica NPs or silica- remains clinical interest in human. As this rapidly expanding area
DNA NPs complexes [134]. of research continues to evolve, it is essential that researchers and
In a pilot study, Praetorius et al. [116] analyzed the transduction clinicians begin to consider and develop mechanisms for the sus-
efficacy of Cy3-labeled silica NPs placed on the RWM of adult tained and targeted delivery of compounds to the cochlea. Applica-
mice. The results demonstrated silica NPs were taken up in inner tion of the device for inner ear drug delivery is perhaps the most
hair cells, vestibular hair cells, spiral ganglion neurons, and sup- demanding of possible uses.
porting cells after RWM delivery, hence yielding an effective cellu- The nanoparticle-based systems have a huge potential for the
lar delivery for the Cy3-labeled silica NPs. Absence of hearing inner ear delivery of various therapeutic agents. The successful
impairment suggested silica NPs were potentially safe for use in the fabrication of NPs has opened a new era for active targeting, which
inner ear. Additionally, the distal part of the central auditory path- will minimize side effects, enable access to tissues of interest, and
way (dorsal cochlear nucleus, superior olivary complex) was found profoundly increase target specificity. The cochlear acceptability of
to be labeled with the Cy3-labeled silica NPs, indicating a retro- the carrier materials is the major hurdle in their successful commer-
grade axonal transport. This study demonstrates that the special cialization. Extensive in vivo studies are required to establish their
material properties of silica NPs can be used to deliver drugs safely in vivo acceptability. The other aspects of NPs such as sterilizing,
to the central auditory system, which is affected by a variety of freeze-drying, shelf life and large-scale industrial production have
processes including aging. However, DNA-silica NPs complexes already been developed to a sufficient standard with the view of
may show a different distribution pattern or even fail to transfect commercialization. Thus, although there is no nanoparticle-based
894 Current Drug Metabolism, 2010, Vol. 11, No. 10 Chen et al.

product for treating the inner ear diseases in the market untill date, [19] Medina, J. E.; Drescher, D. G. The amino-acid content of per-
with the progress that they have seen so far, the day for the mar- ilymph and cerebrospinal fluid from guinea-pigs and the effect of
keted arrival of NPs might not be too far. noise on the amino-acid composition of perilymph. Neuroscience,
1981, 6(3), 505-509.
ACKNOWLEDGEMENTS [20] Scheibe, F.; Haupt, H. Biochemical differences between perilymph,
cerebrospinal fluid and blood plasma in the guinea pig. Hear. Res.,
The work was supported by the National Nature Science Foun- 1985, 17(1), 61-66.
dation of China under contract of 30801553, and the Faculty Con- [21] Thalmann, I.; Kohut, R. I.; Ryu, J.; Comegys, T. H.; Senarita, M.;
struction Funds of Guangdong Pharmaceutical University. Thalmann, R. Protein profile of human perilymph: in search of
markers for the diagnosis of perilymph fistula and other inner ear
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Received: October 12, 2010 Revised: December 23, 2010 Accepted: December 24, 2010