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Studies on a new carrier of trimethylsilyl-modified mesoporous material for controlled drug delivery.

To better control drug delivery rate, a simple and effective approach has been developed for controlled drug delivery carrier system through one-step surface modification of the ibuprofen-impregnated silica MCM-41 with 1, 1, 1, 3, 3, 3-hexamethyldisilazane (HMDS). The 29Si MAS NMR characterization demonstrated that different contents of trimethylsilyl (TMS) groups were successfully grafted onto the samples modified with different silylation times. The results obtained from in vitro tests exhibited that the introduction of TMS groups greatly retarded the ibuprofen release rate. Even after in vitro test for 48 h, only 75% of the impregnated ibuprofen could be released from the modified sample with TMS groups content of 14.5% (related to the total silicon atoms). However, the release of ibuprofen could be completed just after about 1 h from the pure silica MCM-41 under the same release conditions. Furthermore, the release rate of ibuprofen could be well modulated by changing the grafted content of TMS groups, and was found to decrease with increasing grafted amount of TMS groups.

Biological Availability↗

Influence of polyelectrolyte multilayer films on the ICAM-1 expression of endothelial cells.

Recently, the use of polyelectrolyte films has been suggested as a new versatile technique of surface modification aimed at tissue engineering. In the present study, we evaluated the expression of intercellular adhesion molecule (ICAM)-1 of endothelial cells (ECs) seeded on two types of polyelectrolyte multilayer films either terminated by poly(D-lysine) (PDL) or poly(allylamine hydrochloride) (PAH). This work showed that chemical stimulations with tumor necrosis factor (TNF)-alpha induced the ICAM-1 expression of ECs differently depending largely on the film architecture employed. Compared with PAH-ending films, the PDL-ending ones upregulated the ICAM-1 expression of the ECs after a prolonged exposition to TNF-alpha, rendering this film type less favorable in tissue engineering. Cytochalasin D (an F-actin disrupting agent) showed the involvement of the cytoskeleton in the upregulation of ICAM-1 for cells deposited on films terminated by PDL. The PAH-ending films did not perturb the ICAM-1 expression of ECs and might thus enhance the seeding of ECs in vascular engineering.

Cells, Cultured↗

A modified microstamping technique enhances polylysine transfer and neuronal cell patterning.

Macromolecular microstamping with polydimethylsiloxane (PDMS) stamps has been demonstrated to transfer proteins onto glassy substrates for antigen or antibody detection and for cell patterning. For many applications, including neuronal cell patterning, it is important to assure reliable transfer of sufficient quantity of protein. Research has shown that protein transfer is enhanced with the selection of the proper protein-stamp-substrate combination. In addition, detergent studies have shown that detergent-protein complexes detach from surfaces to a greater extent than proteins alone. Therefore, we hypothesized that stamp surface modification (termed here a release layer) can enhance polylysine transfer and benefit cell growth on microstamped substrates. We found unmodified stamps to transfer insufficient polylysine to support good cell survival of hippocampal neurons in a widely used serum-free, reduced-glia cell culture system. However, with modified stamps neuronal growth was reliably good. This enhanced cell growth can be attributed to the increased polylysine transfer due to the release layer rather than increased loading onto the stamp. This enhancement was found to be even greater for two-month old stamps that were stored in water. Furthermore, the physicochemical properties of the release layer can modulate the loading process. Thus, our data supports the conclusions that the release layer: (1) modulates polylysine loading, (2) enhances polylysine transfer, (3) enhances cellular growth on microstamped substrates, and (4) extends the durability (defined as the number of times a stamp can be reused) of PDMS microstamps.

Animals↗

Cationic liposomes modified with non-ionic surfactants as effective non-viral carrier for gene transfer.

A defined change in formulation components affects the physical and chemical characteristics of cationic liposomes (CLs) carriers in many ways. Therefore, a great degree of control can be exercised over the structure by modifying the CLs with various materials, leading to new innovations for carrier improvement. In the present study, surface modifications of cationic liposomes with non-ionic surfactants--sorbitan monoesters serials (Span 85, 80, 40 and 20) were carried out for developing a new gene transfer carrier. Span modified cationic liposomes (Sp-CLs) were prepared by reverse phase evaporation method (RPV) and self-assemble complexes of antisense oligonucleotides/surfactant modifying cationic liposomes were prepared by auto-coacervation through electrostatic effect. Characterization of Sp-CLs and the self-assembled complex was performed by electron microscope, particle size, zeta potential, turbidity and agarose electrophoresis. Furthermore, in vitro cellular uptake experiment showed that Span plays a role in enhancing the cellular uptake of encapsulated oligonucleotides mediated by Sp-CLs by the endocytosis-dependent route. CLs modified with Span 40 significantly facilitated the cellular uptake by COS-7 cells and HeLa cells; also showed some positive effect on gene expression. That suggests it is a potential non-viral carrier for efficient gene transfer.

Cations↗

Adsorption of Acid Blue 193 from aqueous solutions onto DEDMA-sepiolite.

The adsorption of Acid Blue 193 (AB193) onto dodecylethyldimethylammonium (DEDMA)-sepiolite was investigated in aqueous solution in a batch system with respect to contact time, pH and temperature. The surface modification of DEDMA-sepiolite was examined by the FT-IR technique. The pseudo-first-order, pseudo-second-order kinetic models and the intraparticle diffusion model were used to describe the kinetic data and the rate constants were evaluated. The experimental data fitted very well with the pseudo-second-order kinetic model and also followed the simple external diffusion model up to initial 10 min and then by intraparticle diffusion model up to 75 min, whereas diffusion is not only the rate-controlling step. The adsorption capacities of natural sepiolite and DEDMA-sepiolite at pH 1.5 and 20 degrees C were (1.19 and 2.57) x 10(-4) mol g(-1), respectively. The above results indicate that DEDMA-sepiolite has around two times higher adsorption capacity than natural sepiolite. The Langmuir and Freundlich adsorption models were applied to describe the equilibrium isotherms and the isotherm constants were also determined. The Freundlich model agrees with experimental data well. The activation energy, change of Gibbs free energy, enthalpy and entropy of adsorption were also evaluated for the adsorption of AB193 onto DEDMA-sepiolite.

Adsorption↗

Optimization of dye-doped silica nanoparticles prepared using a reverse microemulsion method.

Fluorescent labeling based on silica nanoparticles facilitates unique applications in bioanalysis and bioseparation. Dye-doped silica nanoparticles have significant advantages over single-dye labeling in signal amplification, photostability and surface modification for various biological applications. We have studied the formation of tris(2,2'-bipyridyl)dichlororuthenium(II) (Ru(bpy)) dye-doped silica nanoparticles by ammonia-catalyzed hydrolysis of tetraethyl orthosilicate (TEOS) in water-in-oil microemulsion. The fluorescence spectra, particle size, and size distribution of Ru(bpy) dye-doped silica nanoparticles were examined as a function of reactant concentrations (TEOS and ammonium hydroxide), nature of surfactant molecules, and molar ratios of water to surfactant (R) and cosurfactant to surfactant (p). The particle size and fluorescence spectra were dependent upon the type of microemulsion system chosen. The particle size was found to decrease with an increase in concentration of ammonium hydroxide and increase in water to surfactant molar ratio (R) and cosurfactant to surfactant molar ratio (p). This optimization study of the preparation of dye-doped silica nanoparticles provides a fundamental knowledge of the synthesis and optical properties of Ru(bpy) dye-doped silica nanoparticles. With this information, these nanoparticles can be easily manipulated, with regard to particle size and size distribution, and bioconjugated as needed for bioanalysis and bioseparation applications.

Coloring Agents↗

Cellular basis for failure of joint prosthesis.

In recent times, loosening of joint prosthesis resulting in failure, is of grave concern to orthopedicians. It is estimated that 50% of total hip replacements become loose after 15 years, and most of them require either revision surgery or resection orthoplasty. Neither, newer operative techniques, change in design, use of novel materials, nor surface modifications have helped to circumvent the problem. It is in this context, that attention has been focussed on the role of tissue surrounding the implant, in the loosening of the prosthesis. Tissue response around prosthesis results in either formation of a fibrous layer around the implant, ingrowth into fenestrations on implant or direct bone apposition on prosthesis. Long-term implantation results in implant debris being released into surrounding tissue. These particles initiate a chronic granulonatous inflammation with a significant number of activated macrophages and foreign body type of giant cells, all engaged in attempts to get rid of the debris. These features have been found to be invariably associated with peri-prosthetic lysis of bone. Since such bone resorption is also observed around non-cemented prosthesis, possibly causes other than cement are responsible for the osteolysis. Retrospective studies on failed implants suggest that peri-prosthetic osteolysis is mediated by activated macrophages. Cytokines are capable of stimulating bone resorbing cells, the osteoclasts. Bone resorption results in further loosening of the prosthesis, changes in stress, frictional wear, release of more wear debris and recruitment of more macrophages. Bone death and proliferation of macrophages, thus appear to be the cause for pain and loosening of prosthesis.

Biocompatible Materials↗

A shift in pathway of iron-mediated perchloroethylene reduction in the presence of sorbed surfactant--a column study.

Surface modification of zero-valent iron (ZVI) to enhance its reduction rates for chlorinated ethanes and ethenes has recently attracted great attention. In this research, the enhancement of perchloroethylene (PCE) reduction by ZVI in the presence of sorbed micelles of the cationic surfactant hexadecyltrimethylammonium (HDTMA) was examined in a series of laboratory column tests with varying flow rates and input PCE concentrations. Model simulations using HYDRUS-1D showed that the overall pseudo first-order rate constants for PCE reduction by ZVI increased by a factor of four in the presence of sorbed HDTMA admicelles. The increase in reduction rate was attributed to a higher distribution coefficient (K(d)) for contaminant sorption on surfactant-modified ZVI (SM-ZVI) compared to untreated ZVI. Modeling results also showed that in the presence of HDTMA admicelles 58-100% of PCE reduction occurred via hydrogenolysis. In contrast, only 12-25% PCE underwent hydrogenolysis when HDTMA was absent. The significant increase in TCE production during PCE reduction by SM-ZVI verified a shift in reaction pathway previously observed in batch studies, most likely from beta-elimination to hydrogenolysis. Although this shift in reaction pathway resulted in a higher accumulation of TCE, the combined concentrations of chlorinated hydrocarbons in the effluent were 1.5-5 times lower when SM-ZVI rather than unmodified ZVI was used.

Adsorption↗

Neural differentiation of midbrain cells on various protein-immobilized polyethylene films.

The effect of surface modification of polyethylene (PE) film on differentiation of midbrain (MB) cells obtained from rat embryos was determined by their micromass culture system. When cultured on untreated PE film, cell differentiation was suppressed to approximately two-thirds of that observed in a control culture dish. On the contrary, type I collagen-immobilized PE film increased differentiated foci of the MB cells more than did the untreated PE film. RGDS (Arg-Gly-Asp-Ser) peptide immobilization onto PE film resulted in almost the same differentiation activity as the collagen immobilized PE film. Bovine serum albumin (BSA) immobilization onto PE film enhance the differentiation activity more than did the untreated PE film, but not up to the levels of collagen- and RGDS-immobilized PE. The number of differentiated foci of the MB cells on untreated PE film were increased by the addition of the condition medium prepared from the collagen-immobilized PE film. However, the number of foci was not increased by the addition of other condition media obtained from control dish, untreated, BSA-, and RGDS-immobilized PE. On the other hand, none of these condition media enhanced a differentiation of the neuronal cell line of PC12 cells, suggesting that some factors effectively differentiate midbrain cells, composed of neuronal epithelial and mesenchymal cells, but not the PC12 cells secreted in the condition media prepared from collagen-immobilized PE. In addition, it is probable that neural growth factor was not secreted in these condition media, which could not induce the differentiation of PC12 cells.

Animals↗

Does plasma irradiation improve shear bond strength of acrylic resin to cobalt-chromium alloy?

OBJECTIVES: Plasma treatment leads to surface modification such as the improved wettability of dental materials. Studies have suggested that plasma treatment may bring about an improvement in the shear bond strength between cobalt-chromium alloy and self-curing acrylic resin. METHODS: Forty-eight cobalt-chromium alloy specimens were randomly divided into four groups (each of the groups consisted of 12 specimens); air abrasion, adhesive primer, adhesive primer after air abrasion, or after plasma irradiation. A self-curing acrylic resin, 2 mm thick and 3.6 mm in diameter, was processed on all specimens before storage in distilled water at 37 degrees C for 24 h. Shear bond strength values were determined at a crosshead speed of 0.5 mm/min. Data was statistically analyzed using Scheffé's test. RESULTS: Adhesive primer clearly increased the shear bond strength compared with air abrasion (p < 0.05). The synergy of air abrasion and adhesive primer greatly increased the shear bond strength, showing a five-fold increase over air abrasion, or one and a half-fold over adhesive primer. A similar tendency in increase was observed in the adhesive primer after plasma treatment, but to a lower degree. Adhesive primer after plasma treatment showed half the shear bond strength of the treatment of adhesive primer after air abrasion (p < 0.05). Moreover, adhesive primer after plasma treatment showed a lower shear bond strength than adhesive primer alone although the difference was not statistically significant. SIGNIFICANCE: Plasma treatment does not improve adhesion between cobalt-chromium alloy and self-curing acrylic resin.

Acrylic Resins↗

Preparation and application of a photoreactive thrombin analogue: binding to human platelets.

alpha-Thrombin has previously been shown to bind to specific, saturable glycoproteins on the platelet surface. Modification of the thrombin active site with tosyllysyl chloromethyl ketone (TosLysCH2Cl) does not alter thrombin's binding characteristics. Interaction of alpha-thrombin with high-affinity binding sites (KD = 10(-9) M) initiates the platelet response which involves proteolytic hydrolysis of this glycoprotein. Although TosLysCH2Cl--thrombin binds to and competes for the same sites as alpha-thrombin, it cannot induce platelet stimulation because it is enzymatically inactive. In this study, we describe the preparation and application of photoreactive tritium-labeled thrombin analogues. The alpha-thrombin derivative retains its platelet-stimulating and enzymatic activities and, upon photoactivation, covalently binds to specific platelet membrane components. When freshly washed human platelets are exposed to less than saturation doses (less than or equal to 2 nM) of the thrombin derivatives in the dark and photoactivated, a single labeled complex is detected. The same experiment with greater than saturating doses (greater than or equal to 20 nM) of the thrombin derivative yields a similar complex as well as two additional ones. Molecular weight estimates of these thrombin-bound complexes were obtained by gel filtration and NaDodSO4--polyacrylamide gel electrophoresis. The low dose (high affinity) complex with TosLysCH2Cl--thrombin has an approximate molecular weight of 200 000, while that with active alpha-thrombin is smaller, approximately 120 000, due to enzymatic cleavage. The additional complexes detected with the high thrombin dose had estimated molecular weights of 400 000 and 46 000, respectively, and appeared to be the same for TosLysCH2Cl--thrombin and for the alpha-thrombin coupled platelets. These isolated complexes appear to correspond to the two previously detected populations of thrombin binding sites on the platelet.

Azides↗

Metal chloride primers for bonding dentine with tri-n-butylborane-initiated luting agents.

OBJECTIVES: The bonding of resin to dentine is dependent largely on surface modifications. The purpose of the present study was to test the hypothesis that a role of the metal chlorides used as primers is to initiate polymerization of resin at the resin-dentine interface, thereby affecting bond strength. METHODS: Nine primers were evaluated, comprising aqueous 2-hydroxyethyl methacrylate (HEMA) solutions containing AICl3, CeCl3, CoCl2, CuCl2, FeCl3, NiCl2, MgCl2, SnCl2 or ZnCl2, respectively. One of the two luting agents (Super-Bond resin) consisted of methyl methacrylate (MMA), 4-methacryloyloxyethyl trimellitate anhydride (4-META) and tri-n-butylborane (TBB) initiator. The other luting agent (MMA-TBB resin) consisted of MMA and TBB without 4-META. Extracted bovine teeth were ground to expose the dentine, etched with an aqueous solution of 10% phosphoric acid, primed, and then bonded with stainless-steel rods; tensile bond strengths were determined after 1-day immersion in water. Data were analysed by ANOVA and Duncan's new multiple range test (P < 0.05). RESULTS: Four of the metal chlorides (CuCl2, FeCl3, MgCl2 and ZnCl2) enhanced the bond strengths of MMA-TBB resin to dentine. With Super-Bond resin, maximum bond strengths of 15.6 MPa and 19.0 MPa were recorded with primer containing 2.0 x 10(-5) mol g(-1) FeCl3 and 2.0 x 10(-6) mol g(-1) CuCl2, respectively. CONCLUSIONS: Bonding techniques, combining the use of either cupric primer or ferric primer with 10% phosphoric acid etchant may be suitable for application in seating resin-bonded prostheses with TBB-initiated luting materials.

Acid Etching, Dental↗

Vesicle formed by amphiphilc cucurbit[6]uril: versatile, noncovalent modification of the vesicle surface, and multivalent binding of sugar-decorated vesicles to lectin.

We report a novel vesicle formed by an amphiphilic CB[6] derivative, the surface of which can be easily modified via host-guest interactions by taking advantage of molecular cavities, readily accessible at the vesicle surface, and their strong affinity toward polyamines. Amphiphilic CB[6] derivative 1 synthesized by reaction between (allyloxy)12CB[6] and 2-[2-(2-methoxyethoxy)ethoxy]ethanethiol affords a vesicle that has been characterized by TEM, light scattering, and fluorescent dye entrapment experiments. Treatment of vesicle 1 with FITC (fluorescein isothiocyanate)-spermine conjugate ligand 2, in which spermine serves as a binding motif to CB[6] and FITC as a fluorescent tag, produced a surface-modified vesicle, which can be easily visualized by a confocal microscope. This result provides us with a new noncovalent, modular approach to the modification of vesicle surfaces. By treating the vesicle derived from the amphiphilic CB[6] with a tag-attached polyamine, we can easily decorate the surface of the vesicle with the tag. Sugar-decorated vesicles were prepared by this noncovalent method, and their interactions with concanavalin A (ConA) were studied. The binding constant of the vesicle decorated with mannose-spermidine conjugate 3 to ConA was measured to be approximately 3 x 104 M-1, which is almost 3 orders of magnitude higher than that of free ligand 3 to ConA (K = approximately 50 M-1). On the other hand, the binding constant of the vesicle coated with galactose-spermidine conjugate 4 to ConA was too small to be measured. These results illustrate the specific and multivalent interactions between the mannose-decorated vesicle and ConA. The ability for facile surface modification suggests many practical applications, including its use in targeted drug delivery and immunization.

Bridged-Ring Compounds↗

Influence of surface and protein modification on immunoglobulin G adsorption observed by scanning force microscopy.

Scanning force microscopy has been used successfully to produce images of individual protein molecules. However, one of the problems with this approach has been the high mobility of the proteins caused by the interaction between the sample and the scanning tip. To stabilize the proteins we have modified the adsorption properties of immunoglobulin G on graphite and mica surfaces. We have used two approaches: first, we applied glow discharge treatment to the surface to increase the hydrophilicity, favoring adhesion of hydrophilic protein molecules; second, we used the arginine modifying reagent phenylglyoxal to increase the protein hydrophobicity and thus enhance its adherence to hydrophobic surfaces. We used scanning force microscopy to show that the glow discharge treatment favors a more homogeneous distribution and stronger adherence of the protein molecules to the graphite surface. Chemical modification of the immunoglobulin caused increased aggregation of the proteins on the surface but did not improve the adherence to graphite. On mica, clusters of modified immunoglobulins were also observed and their adsorption was reduced. These results underline the importance of the surface hydrophobicity and charge in controlling the distribution of proteins on the surface.

Adsorption↗

Learning about truth and biases through experience: section surface corrugation, protein denaturation, and staining.

By developing and investigating the use of thin resin sections for electron microscopy, we discovered that many of the current biases became invalid: 1) No evidence could be produced that the involved organic liquids produce the protein denaturation responsible for loss of antigenicity and lowered resolution. For immunolabeling, "water-soluble" resins are not per se to be preferred over less polar and more hydrophobic ones. 2) The relief formed by the corrugated section surface enables access to the antigenic sites. The depth of relief is determined by the strength of copolymerization and depends on temperature, the chemical nature of resin and biological matter, and, thus, also on surface modification through fixatives. The stronger the relief, the better the immunolabeling and the less the image resolution. Strong copolymerization favors flatter relief, but also a hiding of antigenic sites by thin layers of resin. 3) Heavy metal stain remains the main culprit for low resolution of sections when compared to results obtained with other preparation methods.

Image Enhancement↗

Photoreactive analog of peptide FN-C/H-V from the carboxy-terminal heparin-binding domains of fibronectin supports endothelial cell adhesion and spreading on biomaterial surfaces.

The extracellular matrix protein fibronectin (FN) plays an important role in cell adhesion, spreading, and motility. Several cell-adhesion promoting domains exist within fibronectin, and peptide sequences from these domains have been shown to play an important role in cell interactions with fibronectin. Recently, a peptide sequence (FN-C/H-V) from the 33/66 kD carboxy-terminal heparin-binding domains of fibronectin was shown to promote the adhesion and spreading of vascular endothelial cells in vitro. Endothelial cell spreading on this peptide was followed by cytoskeletal reorganization, focal contact formation, and, ultimately, cell migration. In the current study, a photoreactive analog of FN-C/H-V (ASD-V) was generated using a heterobifunctional photoreactive crosslinking agent, sulfosuccinimidyl 2-(pazidosalicylamido) ethyl-1,3'-dithio-propionate. ASD-V was then covalently coupled to polystyrene (PS) and polyethylene terephthalate film (PET) in order to assess the utility of ASD-V for preparing biomaterial surfaces with endothelial cell-adhesion promoting properties. The effects of pre-adsorption time and initial coating concentration on the efficiency of ASD-V coupling to PS and to PET were examined. Contact angle measurements and atomic force microscopy were used to characterize ASD-V-modified surfaces. Finally, the adhesion and spreading of vascular endothelial cells on ASD-V-modified surfaces was assessed. Our results suggest that photoreactive peptides are an effective and convenient means of modifying biomaterial surfaces to impart adhesion-promoting properties and that ASD-V, when coupled to PS and PET, promotes endothelial cell adhesion and spreading and may therefore be useful as a biomaterial surface modification in applications where re-endothelialization is desired (e.g., autologous endothelial seeding of vascular grafts, or transplantation of genetically engineered endothelial cells via polymer-coated stents.

Animals↗

Evaluating sol-gel ceramic thin films for metal implant applications. II. Adhesion and fatigue properties of zirconia films on Ti-6AI-4V.

The degree to which ceramic coatings or thin films applied to bone-interfacing metallic implants can improve the overall performance of these implants with respect to implant fixation, wear, or corrosion relies especially on the response of these films to loading. In this study, the adhesion and fatigue properties of sol-gel zirconia films that could be reproducibly deposited onto polished Ti-6AI-4V substrates was investigated. For zirconia films on the order of 100 nm thick, a shear lag-based strain approach indicated a shear adhesion strength of approximately 275 MPa. Small variations in film thickness and substrate surface preparation had little effect on this adhesion, which was believed to be due to alkoxide molecule interactions with free hydroxyl groups on the substrate surface as well as some limited interfacial diffusion following the 500 degrees C anneal. Subsequent fatigue testing of these films in air using novel tapered rotating beam fatigue samples demonstrated their excellent fatigue characteristics, with films surviving up to 10(7) cycles, the endurance limit of the Ti-6AI-4V (approximately 635 MPa). Overall, the exceptional mechanical properties of this ZrO2/Ti-6AI-4V system along with the inherent advantages of sol-gel processing support continued studies to utilize this technology for implant surface modification.

Alloys↗

Controlling cell interactions by micropatterning in co-cultures: hepatocytes and 3T3 fibroblasts.

The repair or replacement of damaged tissues using in vitro strategies has focused on manipulation of the cell environment by modulation of cell-extracellular matrix interactions, cell-cell interactions, or soluble stimuli. Many of these environmental influences are easily controlled using macroscopic techniques; however, in co-culture systems with two or more cell types, cell-cell interactions have been difficult to manipulate precisely using similar methods. Although microfabrication has been widely utilized for the spatial control of cells in culture, these methods have never been adapted to the simultaneous co-cultivation of more than one cell type. We have developed a versatile technique for micropatterning of two different cell types based on existing strategies for surface modification with aminosilanes linked to biomolecules and the manipulation of serum content of cell culture media. This co-culture technique allowed manipulation of the initial cellular microenvironment without variation of cell number. Specifically, we were able to control the level of homotypic interaction in cultures of a single cell type and the degree of heterotypic contact in co-cultures over a wide range. This methodology has potential applications in tissue engineering, implant biology, and developmental biology, both in the arena of basic science and optimization of function for technological applications.

3T3 Cells↗