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Supramolecular assemblies for the cytoplasmic delivery of antisense oligodeoxynucleotide: polyion complex (PIC) micelles based on poly(ethylene glycol)-SS-oligodeoxynucleotide conjugate
Oishi, M., T. Hayama, et al. (2005), Biomacromolecules 6(5): 2449-54.
Abstract: A novel cytoplasmic delivery system of antisense oligodeoxynucleotide (asODN) was developed by assembling a PEG-asODN conjugate with disulfide linkage (smart linkage) (PEG-SS-asODN) into polyion complex (PIC) micelles through the complexation with branched poly(ethylenimine) (B-PEI). The PIC micelle thus prepared showed a significant antisense effect against luciferase gene expression in HuH-7 cells, far more efficient than nonmicelle systems (asODN and PEG-SS-asODN in free form) and PIC micelle encapsulating the conjugate without the disulfide linkage. Use of poly(l-lysine) (PLL) instead of the B-PEI for PIC micellization led to a substantial decrease in the antisense effect. These results indicate that the PIC micelles formulated from PEG-SS-asODN conjugate and B-PEI is successfully transported from the endosomal compartment into the cytoplasm by the buffering effect of the B-PEI, releasing hundreds of active asODN molecules via cleavage of the disulfide linkage into the cellular interior, responding to a high glutathione concentration in the cytoplasmic compartment. Furthermore, the type of smart linkage (glutathione-sensitive SS linkage vs pH-sensitive linkage) in the conjugates substantially affected the antisense effect of the PIC micelles, depending on the nature of the counter polycation (B-PEI vs PLL).

Supramolecular gels: functions and uses
Sangeetha, N. M. and U. Maitra (2005), Chem Soc Rev 34(10): 821-36.
Abstract: In recent years there has been immense interest in studying gels derived from low molecular mass gelators (supramolecular, or simply molecular gels). The motivation for this is not only to understand the fundamental aggregate structures in the gels at different length scales, but also to explore their potential for futuristic technological applications. Gels have been made sensitive to external stimuli like light and chemical entities by incorporating a spectroscopically active or a receptor unit as part of the gelator molecule. This makes them suitable for applications such as sensing and actuating. The diversity of gel structural architectures has allowed them to be utilized as templates to prepare novel inorganic superstructures for possible applications in catalysis and separation. Gels derived from liquid crystals (anisotropy gels) that can act as dynamically functional materials have been prepared, for example, for (re-writable) information recording. Supramolecular gels can be important in controlled release applications, in oil recovery, for gelling cryogenic fuels etc. They can also serve as media for a range of applications. This tutorial review highlights some of the instructive work done by various groups to develop smart and functional gels, and covers a wide spectrum of scientific interest ranging from medicine to materials science.

Surface analysis methods for characterizing polymeric biomaterials
Merrett, K., R. M. Cornelius, et al. (2002), J Biomater Sci Polym Ed 13(6): 593-621.
Abstract: Surface properties have an enormous effect on the success or failure of a biomaterial device, thus signifying the considerable importance of and the need for adequate characterization of the biomaterial surface. Microscopy techniques used in the analysis of biomaterial surfaces include scanning electron microscopy, transmission electron microscopy, atomic force microscopy, and confocal microscopy. Spectroscopic techniques include X-ray photoelectron spectroscopy, Fourier Transform infrared attenuated total reflection and secondary ion mass spectrometry. The measurement of contact angles, although one of the earlier techniques developed remains a very useful tool in the evaluation of surface hydrophobicity/hydrophilicity. This paper provides a brief, easy to understand synopsis of these and other techniques including emerging techniques, which are proving useful in the analysis of the surface properties of polymeric biomaterials. Cautionary statements have been made, numerous authors referenced and examples used to show the specific type of information that can be acquired from the different techniques used in the characterization of polymeric biomaterials surfaces.

Surface analysis methods of biomaterials used in oral surgery: literature review
Suba, C., N. Velich, et al. (2005), J Craniofac Surg 16(1): 31-6.
Abstract: Titanium is the most frequently used biomaterial in oral surgery because of its positive physical and chemical properties. Clinical studies proved that the properties of titanium can be improved by surface modification techniques. To study the surface of biomaterials, the positive effects of the coatings, the response of the organism (corrosion resistance, physical and chemical stability, the thickness of various coatings, biocompatibility), one must choose and use the adequate analytical method for one's goal. In this article, the authors present the most frequently used analytical methods for the study of the surface morphology and composition of biomaterials. Also, they outline the advantages and disadvantages of specific analytical methods and the field where they are used.

Surface characterization and biological properties study of silicone rubber membrane grafted with phospholipid as biomaterial via plasma induced graft copolymerization
Hsiue, G. H., S. D. Lee, et al. (1998), J Biomed Mater Res 42(1): 134-47.
Abstract: Poly(2-methacryloyloxyethyl phosphorylcholine) (pMPC) was grafted onto the surface of a silicon rubber (SR) membrane (pMPC-SR) by plasma induced grafted copolymerization (PIP). Argon plasma was used to activate the SR surfaces. Determination was also made of the influences of grafted copolymerization reaction time, reaction temperature, and monomer concentration on polymerization yield. The surface properties of SR were characterized by ATR-FTIR, ESCA, and SEM. In those analyses the ATR-FTIR spectra indicated that the pMPC grafted onto the SR surface at 1720 and 3300 cm(-1). The elemental composition and different carbon bindings on the surface of the SR were examined by ESCA. An increasing P1s/C1s value g was obtained in the grafted polymerization yield with a concentration of 0.05-0.5M of MPC in the isolated ethanol solution. The surface morphologies of pMPC-SR differed more than those of control and Ar plasma treated surfaces. The difference could have been caused by the homogeneous graft polymerization of pMPC onto the SR membrane. In the biological analyses, protein adsorption on pMPC-SR surfaces was reduced. The reduced level increased with an increase in the pMPC grafted amount. The epithelial cell attachment and growth onto these samples were suppressed. The blood compatibility for a series of pMPC-SR surfaces was examined by platelet adhesion. Blood platelet morphologies in contact with the high ratio of pMPC-SR surfaces were maintained, meaning that in this case the release reaction for platelets never occurred. Consequently, the high amount of pMPC-SR surface had excellent blood compatibility, further suggesting that prevention of adhesion, activation of platelets, and adsorption of blood protein could be achieved.

Surface characterization of biomaterials by electron spectroscopy for chemical analysis
Ratner, B. D. (1983), Ann Biomed Eng 11(3-4): 313-36.
Abstract: Electron spectroscopy for chemical analysis (ESCA) is perhaps the most valuable single method available for characterizing the surfaces of biomaterials. The ESCA analytical method is rich in information, observes a relevant surface region, and has been shown to generate results that correlate with biological response. In this article, the analysis of actual ESCA data is illustrated by reviewing a study in which polymer-coated glass surfaces, prepared for use as tissue culture substrates, are examined by ESCA. The application of more advanced ESCA techniques such as angular-dependent analysis to this situation is also considered. Finally, published applications of ESCA to the study of polyurethanes, hydrogels, protein films, cell culture substrates, and dental materials are briefly reviewed.

Surface characterization of biomaterials by immunogold staining--quantitative analysis
Park, K., S. R. Simmons, et al. (1987), Scanning Microsc 1(1): 339-50.
Abstract: The labeling of target proteins by immunogold particles has been analyzed based on Einstein's law of Brownian motion. The theory was confirmed from the experiments which employed antifibrinogen gold markers to label fibrinogen molecules adsorbed on the polyethylene surface. The theory predicts that the degree of labeling depends on the concentration of gold markers, temperature, medium viscosity, size of gold markers, and staining time. Of these factors most important is the concentration of immunogold particles. Small change in the marker concentration results in a significant variation in the staining efficiency when other variables are kept constant. The effect of temperature is always accompanied with that of the medium viscosity. There is a linear relationship between the degree of labeling and the temperature when the viscosity effect is combined. The staining of fibrinogen molecules adsorbed on the polyethylene surface at three different temperatures shows a temperature dependence which is in close agreement with the theory. The degree of labeling is inversely related to a square root of the size of gold markers. This analysis makes it possible to maximize the staining sensitivity and to improve the reproducibility of the labeling. Thus, the immunogold staining under a well defined condition allows quantification as well as positive identification and localization of target proteins. This technique has been used to study protein adsorption on biomaterials.

Surface characterization of silver-doped bioactive glass
Verne, E., S. Di Nunzio, et al. (2005), Biomaterials 26(25): 5111-9.
Abstract: A bioactive glass belonging to the system SiO(2)-CaO-Na(2)O was doped with silver ions by ion exchange in molten salts as well as in aqueous solution. The ion exchange in the solution was done to check if it is possible to prepare an antimicrobial material using a low silver content. The doped glass was characterized by means of X-ray diffraction, SEM observation, EDS analysis, bioactivity test (soaking in a simulated body fluid), leaching test (GFAAS analyses) and cytotoxicity test. It is demonstrated that these surface silver-doped glasses maintain, or even improve, the bioactivity of the starting glass. The measured quantity of released silver into simulated body fluid compares those reported in literature for the antibacterial activity and the non-cytotoxic effect of silver. Cytotoxicity tests were carried out to understand the effect of the doped surfaces on osteogenic cell adhesion and proliferation.

Surface charge and the effect of excess calcium ions on the hydroxyapatite surface
Harding, I. S., N. Rashid, et al. (2005), Biomaterials 26(34): 6818-26.
Abstract: This paper describes the use of surface titration as a more direct probe of the surface chemistry of hydroxyapatite (HA) than zeta-potential measurements. The variation in HA surface charge with pH for two different KCl electrolyte concentrations was determined titrimetrically and the point of zero charge (PZC) found to be at a pH of 7.3 +/- 0.1. The curves also demonstrated that HA accumulates positive charge more readily below the PZC than it accumulates negative charge above it. Extended titration data indicated that dissolution occurred more rapidly in increasingly acidic conditions, but was inhibited by increasing electrolyte strength. Similar experiments with 2.5 mM Ca(2+) in the electrolyte show that Ca(2+) adsorption balances loss of H(+) to give a near-neutral surface at any pH above 7 (subject to availability of calcium ions and adsorption sites). The mechanism for adsorption has been shown to be deprotonation of adjacent surface HPO(4)(2-) sites and subsequent adsorption of a calcium ion to the lattice surface site. Furthermore, inhibition of dissolution under alkaline conditions in the presence of Ca(2+) suggested that dissolution was driven by desorption of Ca(2+). Modelling of the adsorption/desorption processes demonstrated that in both pure water and under physiological conditions phosphate groups will predominate at the HA surface. Furthermore, the (200) plane was identified as the likely form of the HA surface. These methodologies and findings are particularly relevant to investigation of biological response with respect to modification of surface hydrophobicity and surface energy or charge.

Surface chemical and mechanical properties of plasma-polymerized N-isopropylacrylamide
Cheng, X., H. E. Canavan, et al. (2005), Langmuir 21(17): 7833-41.
Abstract: Surface-immobilized poly(N-isopropyl acrylamide) (pNIPAM) is currently used for a wide variety of biosensor and biomaterial applications. A thorough characterization of the surface properties of pNIPAM thin films will benefit those applications. In this work, we present analysis of a plasma-polymerized NIPAM (ppNIPAM) coating by multiple surface analytical techniques, including time-of-flight secondary-ion mass spectrometry (ToF-SIMS), contact angle measurement, atomic force microscopy (AFM), and sum frequency generation (SFG) vibrational spectroscopy. ToF-SIMS data show that the plasma-deposited NIPAM polymer on the substrate is cross-linked with a good retention of the monomer integrity. Contact angle results confirm the thermoresponsive nature of the film as observed by a change of surface wettability as a function of temperature. Topographic and force-distance curve measurements by AFM further demonstrate that the grafted film shrinks or swells depending on the temperature of the aqueous environment. A clear transition of the elastic modulus is observed at 31-32 degrees C. The change of the surface wettability and mechanical properties vs temperature are attributed to different conformations taken by the polymer, which is reflected on the outmost surface as distinct side chain groups orienting outward at different temperatures as measured by SFG. The results suggest that a ppNIPAM thin film on a substrate experiences similar mechanical and chemical changes to pNIPAM bulk polymers in solution. The SFG result provides evidence supporting the current theory of the lower critical solution temperature (LCST) behavior of pNIPAM.

Surface energy effects of implant biomaterials on the implant-tissue interface: implications for the rate, character and quality of post-surgical healing
Dorfman, J. D. (1986), J Oral Implantol 12(4): 661-72.

Surface engineering of biomaterials with plasma techniques
Poncin-Epaillard, F. and G. Legeay (2003), J Biomater Sci Polym Ed 14(10): 1005-28.
Abstract: In this study, 3 types of plasma techniques, i.e. plasma modification, plasma deposition and plasma followed by grafting reaction, are used for the fabrication of tools, medical devices and biomaterials. Depending on purpose, bioadhesion of cells and biomolecules is either looked for or avoided. Since the mechanisms of bioadhesion depend on the characteristics of the surface (hydrophilic or hydrophobic), modifying the surface by a treatment will alter the bioadhesion. These treatments are developed for the anti-fouling process, the sterilisation and the improvement of the formation of biofilms. They have also proved useful for the synthesis of biomimicking devices.

Surface engineering of stainless steel materials by covalent collagen immobilization to improve implant biocompatibility
Muller, R., J. Abke, et al. (2005), Biomaterials 26(34): 6962-72.
Abstract: It was shown recently that the deposition of thin films of tantalum and tantalum oxide enhanced the long-term biocompatibility of stainless steel biomaterials due to an increase in their corrosion resistance. In this study, we used this tantalum oxide coating as a basis for covalent immobilization of a collagen layer, which should result in a further improvement of implant tissue integration. Because of the high degradation rate of natural collagen in vivo, covalent immobilization as well as carbodiimide induced cross-linking of the protein was performed. It was found that the combination of the silane-coupling agent aminopropyl triethoxysilane and the linker molecule N,N'-disulphosuccinimidyl suberate was a very effective system for collagen immobilizing. Mechanical and enzymatic stability testing revealed a higher stability of covalent bound collagen layers compared to physically adsorbed collagen layers. The biological response induced by the surface modifications was evaluated by in vitro cell culture with human mesenchymal stem cells as well as by in vivo subcutaneous implantation into nude mice. The presence of collagen clearly improved the cytocompatibility of the stainless steel implants which, nevertheless, significantly depended on the cross-linking degree of the collagen layer.

Surface free energy and interaction of Staphylococcus epidermidis with biomaterials
Ferreiros, C. M., J. Carballo, et al. (1989), FEMS Microbiol Lett 51(1): 89-94.
Abstract: The adhesion of twenty nine Staphylococcus epidermidis strains to teflon, polyethylene, polycarbonate and bovine pericardium was studied in vitro and examined in relation to the surface free energies of both bacteria and biomaterials. All S. epidermidis strains had similar surface free energies, close to that of water, and adhered better to the materials with analogous surface free energies. There was a significant correlation (Kendall's Tau B = 1000) of biomaterial's surface free energy with the number of adhering bacteria. This correlation is inverse (Kendall's Tau B = -1000) when surface hydrophobicity is considered instead of surface free energy. This indicates that in Staphylococcus epidermidis adherence to biomaterials is inversely correlated to the surface hydrophobicity of the last, being so just the opposite of that occurring with other bacteria.

Surface hydroxylation of styrene-butadiene-styrene block copolymers for biomaterials
Sefton, M. V. and E. W. Merrill (1976), J Biomed Mater Res 10(1): 33-45.
Abstract: This work pertains to the development of high strength elastomers potentially useful as nonthrombogenic cardiovascular prostheses. Triblock copolymers of the styrene-butadiene-styrene type have been subjected to surface hydroxylation which provide reactive sites at the surface for the subsequent coupling of heparin while retaining the unique mechanical properties of the SBS copolymers. Curves of hydroxyl content versus the copolymer film thickness demonstrate the effect of swelling in the surface region on the product distribution and on the time dependence of the hydroxylation process. In addition, the effect of time, temperature, and the composition of the reaction bath on the diffusion/reaction process is shown. Finally, the general applicability of this surface modification scheme to the development of biomaterials is discussed.

Surface modification and functionalization through the self-assembled monolayer and graft polymerization
Ruckenstein, E. and Z. F. Li (2005), Adv Colloid Interface Sci 113(1): 43-63.
Abstract: The modification of a surface at the molecular level with precise control of the building blocks generates an integrated molecular system. This field has progressed rapidly in recent years through the use of self-assembled monolayer (SAM) interfaces. Recent developments on surface-initiated chemical reactions, functionalization, and graft polymerization on SAM interfaces are emphasized in the present review. A number of surface modifications by grafting are reviewed. The grafting of polyaniline on a glass surface, previously modified with a silane self-assembled monolayer (SAM), is examined in detail for both planar and 3-D systems, such as fibers, nanoparticles, and even polymer patterned surfaces. We also discuss the graft polymerization of water-soluble polymers on the surface of silicon nanoparticles, which generate stable aqueous colloidal solutions and have numerous applications. Finally, we compare and review some surface-modification techniques on the surfaces of polymers, such as two-solvent entrapment, polymer blending, and chemical grafting, which improve their biocompatibility.

Surface modification and initial adhesion events for intestinal epithelial cells
Hagerman, E. M., S. H. Chao, et al. (2006), J Biomed Mater Res A 76(2): 272-8.
Abstract: Rapid resealing of the mucosal epithelia is imperative following injuries to the small intestine because the mucosa is responsible for the adsorption of nutrients as well as providing a barrier to noxious agents present in the lumen. Tissue engineering may provide a possible solution for treating intestinal erosions, ulcerations, inflammatory bowel disease, and infection. Cell-biomaterial interaction is a critical component in tissue engineering that can determine the success of the tissue construct. Cell-biomaterial interactions can be enhanced by various types of surface modification, which promote integrin ligation leading to increased cell function. In order to relate the effect of surface adhesion molecules to signaling events and macroscopic cell response, an intestinal epithelial cell line, IEC-6, was plated on fibronectin (receptor-mediated) and poly-L-lysine (non-specific) surfaces. Focal adhesion kinase (FAK) phosphorylation, cell spreading, and cell adhesion strength were measured. Results showed increases in FAK phosphorylation generally corresponded to increases in cell spreading and adhesion strength for IEC-6 cells. Therefore, in a simplified system, initial adhesion and signaling mechanisms appeared to correspond to subsequent physical responses in IEC-6 cells relevant to tissue engineering applications. (c) 2005 Wiley Periodicals, Inc. J Biomed Mater Res, 2006.

Surface modification of biodegradable polyesters with fatty acid conjugates for improved drug targeting
Fahmy, T. M., R. M. Samstein, et al. (2005), Biomaterials 26(28): 5727-36.
Abstract: We describe a general method for incorporating target ligands into the surface of biocompatible polyester poly(lactic-co-glycolic acid) (PLGA) 50/50 materials using fatty acids. Avidin-fatty acid conjugates were prepared and efficiently incorporated into PLGA. Avidin was chosen as an adaptor protein to facilitate the attachment of a variety of biotinylated ligands. We show that fatty acid preferentially associates with the hydrophobic PLGA matrix, rather than the external aqueous environment, facilitating a prolonged presentation of avidin over several weeks. We successfully applied this approach in both microspheres encapsulating a model protein, bovine serum albumin, and PLGA scaffolds fabricated by a salt-leaching method. Because of its ease, generality and flexibility, this strategy promises widespread utility in modifying the surface of PLGA-based materials for applications in drug delivery and tissue engineering.

Surface modification of biomaterials through noble metal ion implantation
Buchanan, R. A., I. S. Lee, et al. (1990), J Biomed Mater Res 24(3): 309-18.
Abstract: Studies are described involving effects of noble-metal ion implantation on corrosion inhibition and charge-injection capabilities of surgical Ti-6A1-4V alloy. A major factor linked to excellent long-term biological performance is resistance to metal-ion release to tissues. The elements most resistant to corrosion in aqueous solutions are the noble metals. Disadvantages include expense and general inadequacy of mechanical properties. However, if small quantities can be used to surface-modify a surgical device in the last stage of manufacture, that device could possess an optimum combination of environmental integrity, biological response, mechanical properties, and charge-injection capability at minimum expense. Results for ion-implanted Ir are presented. Iridium has been described as the most corrosion-resistant element known, and its activated oxide as having the highest charge-injection capability of any material known. Ti-6A1-4V samples, ion implanted with 2.5 and 5.0 atomic % peak-maximum concentrations of Ir, were subjected to corrosion treatments to enrich the surface with Ir. Corrosion potential and cyclic voltammetry measurements indicated enrichment in H2SO4, and continued enrichment in isotonic saline, with corrosion potentials approaching that of pure Ir, and charge densities in isotonic saline exceeding that of pure Ir for the 5.0% peak-max Ir implanted material. X-ray photoelectron spectroscopy confirmed the high levels of Ir surface enrichment.

Surface modification of gold and quantum dot nanoparticles with chitosan for bioapplications
Tan, W. B. and Y. Zhang (2005), J Biomed Mater Res A 75(1): 56-62.
Abstract: Gold (Au) and quantum dot (QD) nanoparticles, which have been extensively used in many fields, were encapsulated with a natural polymer, chitosan, to improve their biocompatibility. Characterization was performed using ultraviolet-visible, dynamic light scattering, atomic force microscopy, and transmission electron microscope analyses. It was found that a Au/chitosan ratio of 1:1 and smaller produced chitosan-encapsulated Au nanoparticles of a sufficiently small size, and this result was then applied in the chitosan encapsulation of QDs. The biocompatibility of both types of nanoparticles was assessed in cell culture studies using HT29 human colon carcinoma and NIH 3T3 mouse fibroblast cells. MTT and trypan blue exclusion assays revealed that both chitosan-encapsulated Au nanoparticles and QDs exhibited improved biocompatibility over their bare, nonencapsulated counterparts. Therefore, this study showed that chitosan could be used to encapsulate both Au nanoparticles and QDs in order to enhance their biocompatibility. The approaches developed in this study can also be extended to other nanoparticles for bioapplications as well.

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