270 Current Topics in Medicinal Chemistry, 2008, 8, 270-280

Surface Chemistry Influences Implant Biocompatibility

Paul Thevenot, Wenjing Hu and Liping Tang*
Bioengineering Department, University of Texas at Arlington, PO Box 19138, Arlington, TX 76019-0138

Abstract: Implantable medical devices are increasingly important in the practice of modern medicine. Unfortunately,
almost all medical devices suffer to a different extent from adverse reactions, including inflammation, fibrosis, thrombosis
and infection. To improve the safety and function of many types of medical implants, a major need exists for development
of materials that evoked desired tissue responses. Because implant-associated protein adsorption and conformational
changes thereafter have been shown to promote immune reactions, rigorous research efforts have been emphasized on the
engineering of surface property (physical and chemical characteristics) to reduce protein adsorption and cell interactions
and subsequently improve implant biocompatibility. This brief review is aimed to summarize the past efforts and our
recent knowledge about the influence of surface functionality on protein:cell:biomaterial interactions. It is our belief that
detailed understandings of bioactivity of surface functionality provide an easy, economic, and specific approach for the
future rational design of implantable medical devices with desired tissue reactivity and, hopefully, wound healing
capability.
Keywords: Functional groups, biomaterials, biocompatibility, proteins, cells.

INTRODUCTION the ability to control both (1) the amount and composition of
host protein adhering to a surface, as well as (2) the degree
An aging population, coupled with continued medical
of adsorbed proteins’ conformational changes (exposing
advances, has driven the increased utilization of medical inflammatory epitopes), is the primary goal of modified
implants and demand for new medical devices in the years
biomaterial surfaces.
ahead. Unfortunately, blood-contact medical devices may
trigger a variety of iatrogenic reactions, including surface Many surface modification techniques have been
mediated thrombosis, complement activation, and device- developed during the past twenty years. The modifications of
centered infection [1-2]. There are also significant adverse surface wettablility, hydrophobicity and surface charges have
reactions involved with tissue-contact medical implants. been shown to alter the extent of protein adsorption. Overall,
These include (1) local implant-associated inflammation, increasing surface hyrdophilicity has been shown to improve
including temporomandibular [3-4] and other joint related the biocompatibility of the medical devices at least in vitro.
implants [4-8], (2) phagocyte-mediated oxidation and ”stress Recently more effort has been placed on creating surfaces
cracking” of the implanted materials [9-10], (3) implant uniformly coated with different chemical functionalities in
degradation and tissue fibrosis surrounding mammary an attempt to determine more specifically what surface che-
prosthesis and many other types of implants [11-13], (4) mical entities, beyond hydrophobic or hydrophilic charac-
poor integration of fibrotic tissues artificial ligaments using teristics, can improve biomaterial compatibility. Though
artificial (Woven Dacron®) or biological (such as collagen promising in vitro results have been widely documented in
fiber) materials [14-16], (5) the formation of thick fibrotic literature, surface functionalities have shown little success in
capsules surrounding implants contribute to the failure of exerting a significant influence on tissue responses in vivo.
many types of devices, such as biosensors [17-18], joint The purpose of this article is to review past and recent work
implants [19-20], breast implants [21-22], encapsulated in the area of surface group functional modification of
tissues/cells [23-25], drug delivery systems [26], and eye biomaterials, and what effect these groups have on protein
implants [27-29]. Unfortunately, the mechanisms governing and cellular interactions with surface functionalities.
biomaterial-mediated tissue responses and the possible role
HOW DO BIOMATERIALS TRIGGER FOREIGN
of surface properties affecting such responses are mostly
BODY REACTIONS?
unclear.
At first glance, these inflammatory responses to inert, Commonly used biomaterials are mostly hydrophobic
and have high affinity to a wide variety of proteins. Shortly
non-immunogenic, and non-toxic materials are difficult to
after implantation, biomaterials are covered with a layer of
understand. We and many others have found that, shortly
plasma proteins, predominately albumin, fibrinogen, IgG,
after implantation (minutes to hours), biomaterial implants
fibronectin, and von Willebrand factor [30,32-35]. These
are covered with a layer of host proteins prior to the
adsorbed proteins, possibly via hydrophobic interactions,
accumulation of inflammatory cells [13,30-32]. Thus, it is
generally believed that the composition and state of these tend to assume an altered conformation and to expose
hydrophobic domains which become tightly adherent to
adsorbed proteins play a pivotal role in subsequent host
hydrophobic biomaterial surfaces [36-41]. Such surface-
coagulation and immune reactions [13,30,32-33]. Therefore,
mediated conformational changes of adsorbed proteins have
been demonstrated by several different techniques including
*Address correspondence to this author at the Biomedical Engineering resistance to sodium dodecyl sulfate (SDS) elution [42-43],
program, University of Texas at Arlington, P.O. Box 19138, Arlington, TX scanning angle reflectometry [12,44-45], 'scanning force'
76019-0138; Fax: 817-272-2251; E-mail: [email protected]
microscopy [46-48] and attenuated total reflectance Fourier

1568-0266/08 $55.00+.00 © 2008 Bentham Science Publishers Ltd.

Surface Chemistry Influence Implant Biocompatibility Current Topics in Medicinal Chemistry, 2008, Vol. 8, No. 4 271

transform infrared spectroscopy [37,49]. The conformational interact with immune cells. A similar exposure of these
changes of adsorbed protein are thought to be responsible for RIBS epitopes and fibrin-specific epitopes may also occur
initiating adverse reactions such as inflammation, coagu- upon adsorption and conformational change of fibrinogen on
lation, and foreign body response [39,50-52]. It is believed implantable biomaterials [61-63].
that these protein:biomaterial interactions prompt the Taken together, these findings indicate that upon inter-
exposure of hidden protein structures and sequences that
action with implant surfaces, fibrinogen (i) becomes
serve as receptor sites for inflammatory cells, which then
progressively more adherent, (ii) shows signs of changes in
initiate the foreign body reactions (Fig. 1).
conformation (?denaturation?), particularly involving the D
Although the influence of material surface properties on domain, and (iii) exposes previously occult sequences (e.g.,
subsequent tissue responses is not totally understood, it is RIBS, P1 and P2). Based on these earlier observations, it is
widely documented that polymer surface properties can likely that material properties can be modified to reduce the
affect the amounts and types of bound proteins, as well as extent of neo-epitopes’ exposure and subsequent cell and
the conformation, orientation or binding strength of the tissue responses.
adsorbed protein [12,31,38,53-54]. Adsorbed proteins can
either retain a structure close to that in solution or may INFLUENCE OF SURFACE CHEMISTRY ON
BIOCOMPATIBILITY
conformationally adjust in response to local environments
[55]. This time-dependent conformational readjustment is Surface Modification Techniques
generally believed to affect the cellular responses and
subsequent tissue reactions [12,31,38,53]. In the case of In the search for “perfect” surfaces, material scientists
fibrinogen, the resistance to SDS elution has been correlated have been modifying surface characteristics by a plethora of
with slow, progressive changes in state of the surface protein techniques including physical modifications, chemical modi-
on biomedical polymers such as polyetherurethanes [36]. fications, and radiation [64-65]. In general, the modification
After polyethylene terephthalate (PET) film has been of material surface properties (including chemistry, wetta-
incubated with fibrinogen for 4 hours, more than 60% of the bility, domain composition and morphology) has been shown
protein is resistant to SDS elution [56]. Albumin and IgG to influence protein adsorption and subsequent cellular
behave similarly [57]. A recent study employing grazing responses to biomaterials in vitro. Unfortunately, due to the
angle infrared analysis was conducted to determine the inter- lack of well-defined surfaces (that differ only in one or two
actions of fibrinogen and albumin adsorption onto hydro- properties) and well-characterized animal implantation
phobic and hydrophilic surfaces. Results showed within 1 models, many early studies have produced little insight into
hour of fibrinogen adsorption, changes in secondary the influence of surface properties on the pathogenesis of
structure (amide I band) could be seen suggesting that foreign body reaction. For creating homogenous and well-
changes in fibrinogen structure were due to protein-surface defined surfaces, self-assembled monolayer techniques, che-
interactions [58-59]. The degree of secondary structure mical graft modification, and plasma medication techniques
alteration was shown to be dependent on the properties of the have been developed and used in many recent studies. Each
surface (hydrophilic versus hydrophobic) [60]. It has been method has its unique benefits and drawbacks.
shown that, upon adsorption to plastic tissue culture surfaces Chemical graft modification has been employed to
(which are not precisely analogous to most hydrophobic chemically immobilize compounds onto the surface of a
implant surfaces), fibrinogen changes conformation and biomaterial. This method usually involves covalent conjuga-
exposes multiple receptor-induced binding sites (RIBS). tion of either a protein or monomer to the surface to alter
Two of these RIBS epitopes have been localized to residues surface chemistry, avoiding drawbacks of protein/monomer
gamma112-119 (recognized by mAb 9F9) and A
95-98 detachment by providing long-term stability. The process
(RGDF; recognized by mAb 155B16). Please see Fig. 2 for involves activating the surface with reactive groups followed
the locations of some of these epitopes which are known to by grafting the desired functionality to the surface. Many
Hidden epitopes

Exposed epitopes

Fig. (1). Schematic drawing to depict the protein:biomaterial interactions which often lead to conformational changes of adsorbed proteins
and the subsequent exposure of hidden proinflammatory epitopes.
272 Current Topics in Medicinal Chemistry, 2008, Vol. 8, No. 4 Tang et al.

RGDS
ȕ
AĮ 572-575 Ȗ
Į
P1
Ȗ 190-202

RGDF
Į AĮ 95-98
Ȗ ȕ
P2
Ȗ 377-395
Fig. (2). Schematic drawing of fibrin(ogen) showing the approximate locations of some of the epitopes which have been implicated to be
responsible for triggering foreign body reactions. The potential inflammatory epitopes include P1 (190-202), P2 (377-395), RGDS (A

572-575), and RGDF (A
95-98).

different methods fall into this category and differ only by excited into energetic states by radio frequency, microwave,
the way in which the surface is activated, including chemical or electrons from a hot filament discharge [70]. The process
reactions, UV, radiation exposure, plasma, and ozone provides an economical and effective way to infer func-
exposure [66]. Chemical grafting has been used to mount tionality to a surface, and is compatible with most materials
heparin and PEO to surfaces to increase blood compatibility. currently being investigated in medicine including metals
Drawbacks, especially when grafting proteins, include the [71], polymers [72], and ceramics [73]. One of the great
loss of protein mobility on the surface, and possibly, benefits of plasma modification is the ability to uniformly
presentation of the protein in an unfamiliar conformation on modify surfaces regardless of geometry, allowing for
the surface. Toxic monomer residues may also be left in the modification of micro and nanoparticles [74], films [72], or
grafted surface. Some issues can be addressed by physical 3D components required in tissue engineering and artificial
adsorption techniques, which usually involve dip coating a organs [75]. In addition, the use of pulsed plasmas, in lieu of
material to form a film with desired properties on the the conventional continuous-wave operational mode, pro-
surface. While physical adsorption may help reduce toxic vides an exceptionally high level of film chemistry control-
monomer residues, issues with binding between the materials lability during the coating process [76-80].
arise as well as instability of immobilization when using
An increasing number of chemical functionalities have
proteins.
been placed on implant surfaces. The influence of these
Addressing the drawbacks in chemical and physical functionalities on protein adsorption, cellular and tissue
modification, self-assembled monolayers (SAMs) were reactions has also been investigated. Highlighted in the next
developed as a method to more precisely control the density section, is a summation of the recent understanding of
and conformation of a single or multiple specific functional surface chemistry as related to protein adsorption, cellular
groups on a surface. The general process of producing SAMs and tissue reactions.
is to first activate the bulk material surface, then graft
polymerize onto the activated surface. SAMs can provide Effects of Surface Chemistry on Protein:Surface
flat and chemically well defined surfaces [67], with Interactions
additional benefits of a surface near thermodynamic Surface properties affect the extent of protein adsorption,
equilibrium with closely packed, well-ordered functionalities denaturation, and epitope exposure (specifically 190-202
on the surface [66]. SAMs tend to provide functionalities [P1] and 377-395 [P2]) [38,81]. To reduce unwanted
with control over pattern and densities. Selection of terminal protein:surface interactions, intensive research efforts have
group can also provide a site for further functionalization of been placed on engineering surfaces with uniform functional
the coated surface. Surface functionality via SAMs have groups to “repel” protein. The typical characteristics of
been used to investigate in vivo inflammatory and foreign protein-resistant functional groups include: hydrophilic
body responses of implanted biomaterials [68-69] as well as nature, presence of hydrogen bond acceptors, no hydrogen
many other processes, giving insight into how a particular bond donors, and neutral charge [82], though there appear to
functional group effects a particular process. However, be exceptions. It is generally believed that the identification
SAMs are limited to gold-coated or silver coated surfaces. of protein resistant functional coating can help to tailor mate-
An increasing popular method of altering surface rial surfaces to control protein adhesion and their interactions
functionality of biomaterials is plasma modification. Plasma, following adsorption, and can thus enhance biocompatibility.
which can be regarded as the fourth state of matter, is The effect of surface functionality on fibrinogen adsorp-
composed of highly excited atomic, molecular, ionic, and tion has been investigated vigorously in recent years [83-86].
radical species. It is typically obtained when gases are Computer simulations have show that water is essentially
Surface Chemistry Influence Implant Biocompatibility Current Topics in Medicinal Chemistry, 2008, Vol. 8, No. 4 273

able to successfully compete with fibrinogen over adsorption surface –OH-mediated complement activation, complement
to hydroxyl coated surfaces, reaffirming results indicating fragment C5a-triggered leukocyte activation, and leukocyte
that fibrinogen binds tightly to hydroxyl (-OH) surfaces [84]. adhesion via binding to surface-bound complement fragment
It has also been suggested that an optimum concentration of - C3b [92].
OH groups can enhance the affinity for albumin binding over
fibrinogen [86]. Fibrinogen was however able to hydro- INFLUENCE OF SURFACE FUNCTIONAL GROUPS
ON CELLULAR RESPONSES
phobically interact with methyl functionalized surfaces, in
agreement with previous studies that suggest abundant Substantial research efforts have been placed on studying
adsorption of fibrinogen to methyl (-CH3) functionalized the influence of surface functionality in the cellular response
surfaces [85]. Amine (-NH2) surfaces were able to form to biomaterials. Surface functional groups can influence cell
hydrogen bonds with fibrinogen, tethering it to the amine growth [93-94], likely due to the fact that surface chemical
functionalized surface. In the case of carboxyl (-COOH) functionality affects adsorbed protein and subsequent
groups, fibrinogen was not able to out-compete water to protein:cell interactions. In general, hydrophilic functionality
interact with the carboxyl surface [81]. A similar result was provides low interfacial free energy resulting in reduced
also seen in another investigation in which positive, nega- protein adsorption, cell adhesion, and blood compatibility
tive, hydrophobic, and hydrophilic surfaces were adsorbed [54,95]. It is well established that proteins tend to bind to
with equivalent amounts of fibrinogen. Negatively charged hydrophilic surfaces in a lower amount and less tightly than
surfaces, such as -COOH, formed little to no fibrin compared to hydrophobic surfaces [96-98]. Such reduction of protein
to other surfaces [83]. Further research involving mixed adsorption affects subsequent cellular responses. For
functionalities as well as increased in vivo investigations are example, in an interpenetrating network study, it was seen
needed to develop a more comprehensive understanding of that increasing hydrophilic monomers lead to a decrease in
surface functionality and protein interactions. fibrinogen adsorption and an increase in albumin adsorption
To determine the possible role(s) of functional group [99], as well as a gradual decrease in platelet adhesion [100].
density in modulating protein:biomaterial interactions, we Decreasing platelet adhesion has also been attributed to
have generated surfaces bearing different densities of –OH coatings that are non-ionic and neutral [101-102]. It has also
and –NH2 functional groups using Radio Frequency Glow been suggested that hydrophilic surfaces provide significant
Discharge plasma polymerization technique. As expected, inhibition of leukocyte adhesion and macrophage fusion
increasing the density of surface -OH and -NH2 improves [103], and may result in decreased cytokine secretion and
surface hydrophilicity. In the case of surface:fibrinogen attenuated inflammatory reactions [104]. In contrast, the
interactions, -OH bearing surfaces retained progressively addition of –CH3 groups, imparting hydrophobic functio-
less protein, whereas the -NH2 surfaces adsorbed progres- nality, has been shown to increase leukocyte adhesion [105].
sively more protein as the surface density of -OH and -NH2 The adhesion of leukocytes to solid surfaces depends on
groups increased, respectively. This data confirms that the many different factors such as surface chemistry, charge or
amount of spontaneously adsorbed protein is a function of hydrophilicity, and protein adsorption [105].
surface chemistry. Subsequent studies have been done to Though the influence of surface hydrophobicity on
determine whether the species and density of surface protein adsorption and cellular responses is well docu-
functional groups affect protein conformational changes mented, recent studies have revealed that surface functional
(also called as denaturation). Our results confirm that even groups also affect protein and cell behavior [106-109]. The
these modest changes in surface amine density strongly most common functionalities investigated with relation to
influence fibrinogen adsorption and P1/P2 epitope exposure. biomaterial interactions are the carboxyl (-COOH), hydroxyl
Surface chemistry of biomaterials has also been shown to (-OH), amino (-NH2), and methyl (-CH3) groups. Many
participate in complement cascades through both the recent reports have investigated the effect of these groups on
classical and alternative pathway, leading to the recruitment the binding of adhesive proteins and subsequent cellular
and activation of phagocytes and the adherence and interactions. The results of their findings are summarized
activation of leukocytes [12]. This was first identified in below.
hemodialysis membranes [87] and activation was shown to Carboxyl (-COOH) Functional Group-Bearing Surface
occur on surface with many different functionalities. Results
have suggested that IgG undergoes a more extensive Biomaterial-bearing -COOH displays a negative charged
denaturation on hydrophobic as opposed to hydrophilic functionality on material surfaces [109]. Studies on protein
surfaces, resulting in exposure of active sites which bind adsorption have shown that fibronectin and albumin are
complement proteins [88]. It has also been suggested that the more easily eluted from surfaces coated with –COOH [93].
chemical functionalities themselves can covalently bind This was tied into the fact that this functionality, compared
complement proteins, harboring them from regulatory to other common surface functional groups, adsorbed FN
proteins [89]. Hydroxyl (-OH) functionalized surfaces have and albumin at ratio significantly different to other coatings.
been shown to selectively adsorb IgG from serum, ultimately Surfaces with –COOH also have shown an increase in cell
leading to C3 deposition on the surface [90]. In contrast, growth. A more recent study showed that this phenomenon is
surfaces functionalized with -NH2 and -COOH showed only dependent upon the concentration of –COOH on the surface,
slight activation in comparison to -OH functionalized as an increase in functional group density results in a higher
surfaces [69]. In vivo studies using SAMs with -OH func- negative charge on the surface, which was shown to inhibit
tionality showed increased leukocyte response [91]. This cell growth [94]. Carboxyl functional surfaces, pre-treated
process is composed of at least three continuous steps- the with FN, showed high levels of two FN domains (
51 and
274 Current Topics in Medicinal Chemistry, 2008, Vol. 8, No. 4 Tang et al.

v3) associated with structural and signaling components surface reactions with cells due to the magnitude of tightly
related to focal adhesions [109]. On the other hand, -COOH- bound proteins, and the likelihood that the bound proteins
mediated v3 exposure has been shown to inhibit osteoblast will expose sites responsive to inflammatory cells.
differentiation and mineralization. Similarly, increase in v3
Surfaces with Mixed Functionality
integrin expression on –COOH surfaces was correlated with
a low level of myoblast differentiation though cell prolife- In the recent years, research has progressed to the
ration levels were high [110]. evaluation of mixed functional group chemistries on
surfaces. The goal of these studies is to attempt to combine
Hydroxyl (-OH) Functional Group-Coated Surfaces
favorable properties of both functionalities onto one surface
The hydroxyl group functionality (-OH) represents a to enhance biocompatibility. Interestingly, the properties of
neutral, hydrophilic surface. Early research into surface each functional group seem to almost equally contribute to
functionality suggested that an increase in oxygen containing the overall properties of the surface. Using a SAM
functionalities was proportional to cell growth [111], combining -NH2 (positive charge) and -COOH (negative
sparking further research in oxygen containing functional charge) functionality in different proportions, it was seen
groups such as –OH. FN adsorbed onto -OH functional that at equal molar fractions the lowest platelet adsorption
groups (in comparison with –CH3 functional group) show was seen [119]. This equal proportion of the functionalities
high levels of 51 levels leading to increased cell adhesion left the surface with a near neutral charge. This finding
strength as well as increased levels of structural signaling highlights the possible importance of surface neutrality
components related to focal adhesions [109]. Further related to blood compatibility of biomaterials.
investigation with osteoblasts showed high levels of
Another study, investigating -OH and -CH3 mixed
differentiation and mineralization with –OH functionality as
SAMs, found a decrease in platelet adhesion and activation
opposed to other functional groups. Rather contradictory, -
as well as decreased fibrinogen adsorption with increasing
OH functionality has been shown to have reduced plasma hydrophilicity, attributed to the -OH functionality [120].
protein adsorption and thus higher platelet compatibility
This study further highlights the importance of a wettable
[112]. It has been suggested that the charge neutrality and
surface in decreasing blood platelet interactions. With
hydrophilic nature of the -OH functionality has low protein
regards to surface adhesion, it was seen that hydrophobic
affinity, and thus protein repelling properties [113].
SAMs combining -COOH and -CH3 groups had a higher
Amine (-NH2) Functional Group-Rich Surfaces adhesion than hydrophilic -COOH and -OH SAMs [121]. A
study using -PO3H2 functionality with -CH3 showed that
Amine group (-NH2) functionality displays a positive
though total number of platelets increased with increasing -
charge to the biomaterial surface. Studies using FN and
PO3H2 content, the number of activated platelets decreased
osteopontin (OPN) show favorable protein conformations
[122]. However, when considering -PO3H2 and -OH mixed
after adsorption to the positively charge -NH2 surface
SAMs, the number of activated platelets increased with
[109,114]. Particularly, -NH2 surfaces promote the exposure
increasing -PO3H2 with the lowest adhesion seen at almost
of high density bound receptors as well as focal adhesion equivalent proportion of the functionalities. Highlighting the
components by adsorbed fibronectin [109]. These favorable
importance of the fact that OH functionality has been shown
protein adsorption profiles often lead to increased endo-
to have higher platelet compatibility than -CH3 due to its
thelial cell growth [114] and enhanced differentiation and
hydrophilic nature and neutral charge. This result also
mineralization of osteoblast cells [109,115]. Preferable
suggests that the platelet compatibility of -PO3H2 (low
adhesion, growth, and matrix formation was also shown with
hydrophilic and anionic) is somewhere between -OH and -
fibroblasts on –NH2 surfaces compared with other coatings CH3 [122]. At this time, there has been no work published
[116-117]. Further analysis showed than within 45 minutes,
with mixed SAMs in vivo. The in vivo interactions of mixed
cells cultured on -NH2 had already begun the formation of
surface functionality would likely provide valuable
focal adhesion plaques, linked to increased cell spreading on
information and fill a large information void on the effects of
the surfaces [116]. However, in a study using myoblasts, -
surface functionality in vivo.
NH2 functionality was shown to promote high levels of
myoblast proliferation with low levels of differentiation EFFECT OF SURFACE FUNCTIONALITY ON
[110]. TISSUE RESPONSES
Methyl (-CH3) Functional Group-Bearing Surfaces Despite a significant body of in vitro results, there are
very few works which had focused on the effects of surface
The -CH3 group is the major component of commonly
functional groups in vivo. Almost all in vivo studies were
used polymeric materials and provides a hydrophobic sur-
carried out in an acute implantation model. The results from
face on biomaterials. It is generally accepted that the hydro-
phobic –CH3 functionality promotes protein adsorption, those works have supported that surface functionality may
alter biomaterial-mediated acute inflammatory responses in
usually in conformations unfavorable for desired cellular
vivo [108,123-124]. Using an intraperitoneal implantation
interactions [109]. As a result, -CH3 functionality has also
model with RFGD functionalized disks, our early results
shown increased fibrinogen binding, platelet accumulation
have shown that surface functionality prompts different
and thus poor blood compatibility [112]. A study measuring
extents of acute inflammatory responses in the order: -NH2 >
the adhesion strength of fibrinogen, albumin, and IgG
showed that all three proteins adhered with the highest CH3
-CFx > -OH > siloxyl group [108]. A recent study was
carried out using using SAMs to produce hydrophobic –CH 3
strength to -CH3 bearing surfaces [118]. Overall, these re-
and hydrophilic -OH and -COOH surfaces. The results have
sults suggest that -CH3 surfaces will likely have unfavorable
Surface Chemistry Influence Implant Biocompatibility Current Topics in Medicinal Chemistry, 2008, Vol. 8, No. 4 275

shown that - CH3 surface functionality slightly increases the involving tissue engineering scaffolds is focused on the
thickness of the fibrous capsule formed around implants as addition of ECM derived materials to enhance cell growth
well as higher recruitment of inflammatory cells in and differentiation (126). Surface functional groups are
comparison to -OH and -COOH functionalities [68]. In a being used primary to facilitate the chemical addition of
separate study it was seen that –CH3 terminated SAMs lead specific membrane ligands to promote cell adhesion. As
to an increase in leukocyte and phagocyte migration to the knowledge of the in vivo effects of surface functional group
implant site compared to the gold surface alone [124]. coating of biomaterial surfaces expands, we will likely see
increased research and application of surface group
Surprisingly, almost all previous studies failed to show
functionalized scaffolds.
the effects of surface chemistry on chronic fibrotic responses
in vivo [68,108,125]. We speculated that the relatively Interestingly, the results from recent studies suggest that
ineffectiveness of surface functionality on long-term in vivo surface functionality can induce cell differentiation. A study
cellular responses may simply reflect insufficient cell:surface using Caco-2 cells showed that there was a relationship
interactions. Specifically, in an in vitro setting, cells are between perfluoronated functional coating and degree of cell
seeded on top of functional group and surface functionality, differentiation [127]. In a separate study, –COOH-mediated
thus, exerting maximal effect on cellular responses. On the
v3 integrin exposure has been shown to inhibit osteoblast
other hand, in an in vivo environment, the functional groups differentiation and mineralization [112]. Investigation of -
on non-porous implants can only interact with the first layer OH functionality with adhered osteoblasts showed high
of the cells which represent only a small portion of the levels of differentiation and mineralization with –OH, and -
surrounding cells/tissue. These relatively poor cell-functional NH2 functionality as opposed to other functional groups
group interactions thus minimize the effect of surface including -COOH, -SO4, and –CH3 terminal groups [109,
functionality on cellular responses. Thus we hypothesized 112,115]. There remain many possibilities as to the characte-
that possibly the influence of implant surface chemistry may rization of protein adherence to functional coatings and the
be substantially enhanced by increasing the interaction subsequent effects of these proteins and their configuration
between functional groups and cells in applications such as on cell differentiation.
tissue scaffolds and drug releasing microspheres. To test this
hypothesis, we have examined, in vivo, responses to FUNCTIONALIZED SURFACES TO IMPROVE
DRUG DELIVERY
microspheres bearing different, molecularly tailored, surface
functionalities. Spherical particles were chosen for this Surface Chemical Effect on Reducing Diffusion Barrier
purpose to maximize the surface to volume ratio of the and Improving Drug Release
implants and also to mimic microspheres used for controlled
drug release. To test this hypothesis, we have recently Like many other implantable medical devices, drug
reported a study using functionalized micron-sized particles, release implants are often surrounded by thick layer of
created and tested for their ability in modulating tissue collagenous fibrotic tissue a few days (weeks) after implan-
responses to biomaterial implants. In this work, the surfaces tation. The fibrotic tissue may serve as diffusion barrier and,
of polypropylene particles were controllably coated with thus, reduce the drug transport from implanted drug release
four different functional groups, specifically –OH, -NH2, - devices to the circulation (Fig. 3). Therefore, it is generally
CFx and –COOH, using a RFGD plasma polymerization believed that the reduction of biomaterial-mediated fibrotic
technique [74]. The effect of these surface functionalities on reactions may improve the drug release properties [128-130].
host tissue responses were then evaluated in vivo. Surfaces Results using SAMs on gold have shown that implants
with hydrophilic –OH and –NH2 surface groups induced the coated with -COOH and -OH functionality produce thinner
thickest fibrous capsule accompanied with the greatest fibrous capsules than -CH3 functionality [68]. Results from
cellular infiltration into the implants. As previously men- our laboratory using polypropylene functionalized micros-
tioned, hydrophilic groups have been shown to reduce pro- phere implants also showed thin capsules for -COOH
tein adsorption, though these studies used thin films consis- functionality. However, significantly increased fibrous
ting of functionality derived from SAMs. In contrast, surfa- capsule formation was found surrounding implants with -OH
ces with –CFx and –COOH exhibited the least inflam- functionality [74]. More research is needed to determine the
matory/fibrotic responses and cellular infiltrations. The relationship between surface functional groups and resulting
results suggest the surface/volume ratio of the implant, as drug release properties.
well as surface functionality, may play an important role in Surface Functionality on Improving Drug Cellular
influencing tissue responses and biocompatibility [74]. Uptake
ROLE OF SURFACE CHEMISTRY IN TISSUE A recent study has also suggested that surface
ENGINEERING functionality of drug delivery vehicles affects both the rate
Investigations into the effects of surface functionality in and mechanism of cell uptake. Using dendrimers with
tissue engineering are still in its infancy. Recently groups terminal groups of -NH2, -OH, and PEG, results have shown
have focused on the incorporation or modification of that -NH2 surfaces were able to enter cancer cells at a higher
scaffold surfaces to enhance hydrophilicty. As previously rate than -OH and PEG functionalized surfaces. It is
mentioned, knowledge of how functional groups effect cell suggested that the favorable interactions of the cationic -NH2
adhesion properties can be used to modify scaffold surfaces surface with the cell membrane allow quicker passage of
to enhance cell adhesion while also reducing inflammatory drug delivery vehicles into the cell. It is likely that the -OH
response to the scaffold material. Currently most research and PEG functionalities, due to their anionic nature, increase
affinity to the cell membrane, and then taken in by an
276 Current Topics in Medicinal Chemistry, 2008, Vol. 8, No. 4 Tang et al.

Fibrotic Drug release particles

Fig. (3). Prominent fibrotic tissue formation was found surrounding poly-L-lactic acid particles which were subcutaneously implanted in
Balb/C mice for 2 weeks. The presence of collagenous fibrotic tissue create diffusion barrier for drug transport from implanted particles to
blood stream.
Table 1. Effect of Surface Functionality on Protein Adsorption, Cell Behavior and Tissue Responses

Surface Functional Group Protein, Cell, and Tissue Properties

Carboxyl (-COOH) • Preferentially interacts with fibronectin and albumin [93].

hydrophilic, negatively • Enhances fibronectin adsorption and exposure of cell adhesive domains related to focal adhesion and cell
charged growth [109].
• Increases nanoparticle uptake [139].
• Attenuates inflammatory responses and reduces fibrotic capsule formation in vivo [74].

Hydroxyl (-OH) • Reduces plasma protein affinity [113].

hydrophilic, neutral charge • Promotes fibronectin to expose cell adhesive domains related to focal adhesion [109].
• Enhances differentiation and increase mineralization of osteoblasts [109].
• Thickened fibrotic capsule with high levels of cell infiltration in vivo [74].

Amine (-NH2) • Possesses high fibronectin affinity leading to increased endothelial cell growth [114], differentiation and
hydrophilic, positive charge mineralization of osteoblasts [115], and myoblast proliferation [110].
• Promotes focal adhesion formation, cell spreading, and matrix formation with fibroblasts [116].
• Increases particle uptake [137].
• Triggers acute inflammatory responses, thick fibrotic capsule formation, and cell infiltration in vivo [74,108].

Methyl (-CH 3) • Tightly binds fibrinogen leading to platelet accumulation [112].

hydrophobic, neutral charge • Binds IgG more tightly than other surface functional groups [118].
• Leads to thick fibrotic capsule formation and high recruitment of inflammatory cells in vivo [68].
• Increased leukocyte adhesion and phagocyte migration [124].

Mixed -NH2 & -COOH • When mixed at equal molar fractions (near neutral charge), platelet adsorption is at its lowest compared to
positive & negative other fractional mixes of the functionalities [119].

Mixed -OH & -CH3 • As the fraction of -OH increases, platelet adhesion and fibrinogen adsorption decrease [120].
hydrophobic & hydrophilic

Mixed -PO3 H2 & -OH • Equal molar fractions of -PO3H 2 and –OH had the lowest number of adherent platelets [122].
negative & neutral

endocytotic route [131]. Another study investigating -OH, Surface coating has been shown to effect cellular uptake
-NH2, and -COOH terminal groups on dendrimers showed of nanoparticles [133-134]. In general, micro and nano-
that -COOH and -OH functional surfaces tend to have particles with a hydrophilic shell show decreased protein
increased residence times in vivo, which may be attributed to adsorption and increased circulation time [135]. Particles
their ability to resist recognition by the body through protein modified with a PEG shell, with a hydrophilic –OH surface
adsorption, as well as cell uptake properties due to surface functionality, can reduce particle uptake by mononuclear
functionality [132]. phagocytes [136]. A recent study, examining the effects of
altering surface charge on particle uptake, showed that
Surface Chemistry Influence Implant Biocompatibility Current Topics in Medicinal Chemistry, 2008, Vol. 8, No. 4 277

increasing –NH2 group functionality on the particle surface [5] Thornhill, T. S.; Ozuna, R. M.; Shortkroff, S.; Keller, K.; Sledge,
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[138]. A recent study has shown that -COOH functional by inflammatory cells derived from the joint capsule of hip
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A significant body of research clearly demonstrates that Klein, M.; Prantl, L. Does the surface structure of implants have an
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ACKNOWLEDGEMENTS artificial ACL reconstruction. Knee Surg. Sports Traumatol.
Arthrosc. 1993, 1, 9-12.
This work was supported by a NIH grant RO1 [16] Prescott, R. J.; Ryan, W. G.; Bisset, D. L. Histopathological
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GM074021, a Texas Higher Education Coordinating Board’s J. Clin. Pathol. 1994, 47, 375-376.
Advanced Technology Program (003594-0003-2006), and an [17] Turner, R.F.; Harrison, D.J.; Rajotte, R. V. Preliminary in vivo
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