Surface modification of titanium substrate with a novel covalently-bound copolymer thin film for improving its platelet compatibility.

Despite of its widely uses in various clinical applications, the titanium-based material still faces different challenges, such as hemocompatibility and anti-biofouling characteristics required in various situations. The objective of this investigation was to develop a novel surface modification strategy for titanium-based material to improve the platelet compatibility that is important in rigorous blood-contacting cardiovascular applications. In this work, a series of copolymers, which composed of novel 6-acryloyloxy hexyl phosphonic acid (AcrHPA) and sulfobetaine methacrylate (SBMA) was synthesized. The phosphonic acid group in these copolymers can impart covalent binding to the titanium substrate while the zwitterionic sulfobetaine functionality is considered being able to reduce the platelet adhesion and activation on the modified titanium substrate. NMR analyses suggested that copolymerization reaction is likely not an ideal statistical reaction but to add the monomers in a random order. Studies have shown that the composition of the monomers affected the surface characteristics and platelet compatibility of these covalent-bound AcrHPA-SBMA copolymers on titanium substrate. Contact angle analysis has shown the addition of SBMA can increase surface hydrophilicity of the spun-coated copolymers. In addition, AFM analyses have revealed that the surface roughness of the spun-coated copolymer layer were varied with the ratio of AcrHPA and SBMA. The most platelet compatible surface was noted on the one modified by the highest amount of SBMA added (i.e. 70 mol%) in copolymerization. In summary, the surface modification scheme presented here would be of potential as well as manufacturing process applicable for future development in blood-contacting titanium-based biomedical devices.

Application of near-infrared-to-blue upconversion luminescence for the polymerization of resin cements through zirconia discs.

OBJECTIVES: To investigate a near-infrared-to-blue luminescence upconversion curing method for polymerizing resin cements under zirconia discs. METHODS: Lava zirconia discs of different thicknesses (0.5-2.0 mm) were manufactured. First, the transmittances of the NIR and two blue lights (BLs) (LED and halogen lights) through these discs were measured. Second, NaYF4:Yb3+/Tm3+ upconversion phosphor (UP) powder was milled into 0.5-µm particle sizes. A light-curable resin cement VariolinkII base was chosen as the control (UP0), and an experimental cement (UP5) was prepared by adding 5 % UPs. These two cements were examined using multiphoton excitation microscopy for particle distribution. UP5 and UP0 were polymerized with or without zirconia shielding then subjected to a microhardness test. A multifold analysis was performed to examine the effects of zirconia thickness, curing protocols (pure BL or combined BL and NIR curing), and cement type. RESULTS: The transmittance of NIR was superior to that of BL through zirconia discs of all thicknesses. UP particles were homogeneously distributed in UP5 and emitted blue luminescence under 980-nm NIR excitation. UP5 showed higher microhardness values than UP0 under any curing protocol or zirconia shielding condition. The combination of 20-s BL and 40-s NIR curing yielded the highest microhardness in uncovered UP5. However, combining 40-s BL and 20-s NIR curing surpassed the other groups when the zirconia discs were thicker than 0.5 mm. SIGNIFICANCE: NIR exhibits higher transmission through zirconia than BL. UP particles work as strengthen fillers and photosensitizers in cements. NIR upconversion curing could be a new strategy for polymerizing resin cements under thick zirconia restorations.

Investigations of silane-MDP interaction in universal adhesives: A ToF-SIMS analysis.

OBJECTIVES: The purposes of this study were to investigate whether the presence of silane in universal adhesives affects the functions of 10-methacryloyloxydecyl dihydrogen phosphate (MDP) and adhesion to zirconia. METHODS: Two silane-containing universal adhesives (Scotchbond Universal (SBU) and Clearfil Universal-Bond (CUB)) and two silane-free adhesives (All-Bond Universal (ABU) and SE-Bond primer (SE)) were individually applied on zirconia disks. Time-of-flight secondary-ion-mass-spectrometry (ToF-SIMS) examined the distributions of MDP- and silane-related ions, as well as evidence of zirconium phosphate (ZrP) compounds, on the surface and interfacial regions using a depth profiling mode. The hydrophilicity and resin wettability of the treated zirconia were examined using a contact angle test. For the shear bond strength (SBS) test, the zirconia disks were air-blasted, treated with the assigned adhesives, and bonded with pre-cured composite cylinders using a resin cement. These resin-zirconia assemblies received a bond test after 24-h storage. RESULTS: Both SBU and CUB exhibited silane-related ions and ZrO2(OH)-, but fewer PO- ions in the interfacial regions. CUB had more siloxane-related ions. SE-treated zirconia had abundant PO- ions and particularly high PO3-- and ZrP- related ions in the interfacial regions. The silane-free adhesives exhibited a higher affinity to both water and adhesive liquids. SE showed significantly higher SBSs compared to ABU, while SBU and CUB were not statistically different. SIGNIFICANCE: The silane content may cause hydroxylation of zirconia and affect MDP adsorption. An acidic pH accelerated the condensation of silanol. The bond performance of the MDP-based adhesive could be influenced by the silane content and other components.

Interaction of silane with 10-MDP on affecting surface chemistry and resin bonding of zirconia.

OBJECTIVE: To investigate the effect of silane contents on their chemical interaction with 10-methacryloyloxydecyl-dihydrogen phosphate (MDP), and affecting the bonding of MDP to zirconia by time-of-flight secondary ion mass spectrometry (TOF-SIMS) and solid-state nuclear magnetic resonance (SSNMR) spectroscopy. METHODS: Zirconia (Cercon ht, Dentsply) slabs were prepared and fully sintered. Experimental primers SE-5 and SE-10 were formulated by adding 5 wt% and 10 wt% γ-methacryloxypropyltrimethoxysilane to an MDP-based primer SE BOND (SE), respectively. SE, SE-5, and SE-10 were applied on the assigned zirconia slabs. The chemical compositions on the surface and adhesive interfaces were examined by TOF-SIMS in a depth-profiling mode. Hydrophilicity and resin affinity of treated zirconia were analyzed. The bond strengths to resin cylinder were examined either after 24-h storage or thermocycles. In addition, zirconia powders treated with three primers were assessed by SSNMR spectrometry for the adsorption of MDP. RESULTS: TOF-SIMS analysis showed that SE treatment generated the greatest amount of P-O-Zr related ions, which reduced in SE-5 and SE-10 groups. The 3D ion-images illustrated the generation of ZrO2(OH)- ions with silane contents. The SSNMR analysis revealed that the chemical bonding was mainly P-O-Zr ionic bonds in SE but shifted to P-OH-Zr hydrogen bonds in SE-5 and SE-10. SE-5 and SE-10 treated zirconia presented higher hydrophilicity and affinity to resin compared to Zr did. SE showed the highest initial bond strength which significantly decreased after thermocycling. SIGNIFICANCE: MDP adsorption onto zirconia via P-O-Zr ionic bond promotes bonding with resin. The silane enhances the hydroxylation of zirconia and impairs the adsorption of MDP, but does not adversely affect the bond durability.

Improving the surface biocompatibility with the use of mixed zwitterionic self-assembled monolayers prepared by a proper solvent.

In this study, the mixed self-assembled monolayers (SAMs) containing the mixture of long-chain alkanethiol, SH(CH(2))(11)NH(2) and SH(CH(2))(10)SO(3)H, was prepared as a model surface to examine the interaction between the biological environment and artificial surface. The 10% (v/v) NH(4)OH ethanolic solution and DMSO were chosen as the solvents for the preparation of these mixed SAMs and the "solvent effect" was discussed. X-ray photoelectron spectroscopy (XPS) has indicated that -SO(3)H/-NH(2) mixed SAMs formed from 10% (v/v) NH(4)OH ethanolic solution were surface "-SO(3)H poor", while a nearly equivalent amount of surface -SO(3)H functionality was presented on the mixed SAMs formed from DMSO. This has resulted from the different solvation capability between solvent molecules and the alkanethiol. Such solvent effects were also reflected in various surface properties such as surface wettability and surface zeta potential. The mixed SAMs formed from DMSO were more surface hydrophilic and less negatively surface charged than from 10% (v/v) NH(4)OH ethanolic solution. In addition, these mixed SAMs formed from DMSO exhibited the least amount of protein adsorbed as well as a better platelet compatibility than its counterpart from 10% (v/v) NH(4)OH ethanolic solution. These findings indicated that choosing a proper solvent for mixed zwitterionic SAM can greatly affect its surface properties and biocompatibility, such as to form a surface with near neutrality for reducing protein adsorption and subsequent platelet adhesion and activation.

Effect of alkyl chain length and fluorine content on the surface characteristics and antibacterial activity of surfaces grafted with brushes containing quaternized ammonium and fluoro-containing monomers.

Although considerable efforts have been made to vary the alkyl chain length in the quaternary ammonium compounds (QACs) for optimizing the antibacterial activity, only few researchers have systematically investigated the combinatory effects of alkyl chain length and another acryl monomers with the different chemical configuration on the antibacterial activity of the modified substrate. In this study, by surface grafting of various copolymeric brushes, different modified cotton substrates were prepared by surface-initiated reversible addition-fragmentation chain transfer (RAFT) polymerization reaction for exploring the effects of alkyl chain length of QACs and the fluorine content on antibacterial and anti-microbial adhesion characteristics. The quaternized monomers used were prepared by quaternization of 2-(dimethylamino) ethyl methacrylate (DMAEMA) with 1-bromooctane (DMAEMA + 8), and 1-bromopropane (DMAEMA + 3). The fluoro-containing monomer was 2,2,2-Trifluoroethyl methacrylate (TFEMA). Ethyl methacrylate (EMA) was also used for comparison. Results have shown that the optimal antibacterial and anti-microbial adhesion characteristics were noted on the substrates grafted with DMAEMA + 8 and TFEMA. This can be attributed to the enhanced degree of surface quaternization due to the hydrophobic interactions between the grafted TFEMA and DMAEMA + 8 chains, leading to an increase in antibacterial efficacy of modified cotton substrates.

Novel Polymerization of Dental Composites Using Near-Infrared-Induced Internal Upconversion Blue Luminescence.

Blue light (BL) curing on dental resin composites results in gradient polymerization. By incorporating upconversion phosphors (UP) in resin composites, near-infrared (NIR) irradiation may activate internal blue emission and a polymerization reaction. This study was aimed to evaluate the competency of the NIR-to-BL upconversion luminance in polymerizing dental composites and to assess the appropriate UP content and curing protocol. NaYF4 (Yb3+/Tm3+ co-doped) powder exhibiting 476-nm blue emission under 980-nm NIR was adapted and ball-milled for 4-8 h to obtain different particles. The bare particles were assessed for their emission intensities, and also added into a base composite Z100 (3M EPSE) to evaluate their ability in enhancing polymerization under NIR irradiation. Experimental composites were prepared by dispensing the selected powder and Z100 at different ratios (0, 5, 10 wt% UP). These composites were irradiated under different protocols (BL, NIR, or their combinations), and the microhardness at the irradiated surface and different depths were determined. The results showed that unground UP (d50 = 1.9 µm) exhibited the highest luminescence, while the incorporation of 0.4-µm particles obtained the highest microhardness. The combined 20-s BL and 20-120-s NIR significantly increased the microhardness on the surface and internal depths compared to BL correspondents. The 5% UP effectively enhanced the microhardness under 80-s NIR irradiation but was surpassed by 10% UP with longer NIR irradiation. The combined BL-NIR curing could be an effective approach to polymerize dental composites, while the intensity of upconversion luminescence was related to specific UP particle size and content. Incorporation of 5-10% UP facilitates NIR upconversion polymerization on dental composites.

Studies of zwitterionic sulfobetaine functionalized polypropylene surface with or without polyethylene glycol spacer: surface characterization, antibacterial adhesion, and platelet compatibility evaluation.

Microbial adhesion reduction as well as platelet compatibility improvement have been suggested as the key requirements for developing blood-contacting synthetic biomaterials. Surface grafting of hydrophilic polyethylene glycol chains or alkyl chains with zwitterionic terminal ends has been proposed for reducing microbial or platelet adhesion. Nonetheless, none has been reported to incorporate both polyethylene glycol and zwitterionic terminal functionality on the same surface-grafted alkyl chain. In this investigation, a novel surface modification scheme was reported for grafting zwitterionic alkyl chains with or without polyethylene glycol as the spacer. It was noted the bacterial adhesion reduction capability on the zwitterionic modified surface was dependent upon the use of polyethylene glycol spacer or not and the strain of microbe tested. Besides, the zwitterionic modified ones all showed greater antimicrobial adhesion capability than the surface modified with polyethylene glycol alone. On the other hand, significantly reduced platelet adhesion and activation were found, but with no statistical differences noted among the polyethylene glycol-modified surface and zwitterionic ones, with or without polyethylene glycol spacer. These suggested that the use of polyethylene glycol spacer on the zwitterionic terminated surface could further enhance the antimicrobial adhesion against gram-negative bacterial while still keeping its platelet compatibility.

Studies of polypropylene surface modified with novel beta-thiopropionate-based zwitterionic polymeric brush: synthesis, surface characterization, and significantly reduced fouling characteristics evaluation.

Creating a surface with anti- or reduced fouling characteristics can lead to a reduction in nonspecific protein adsorption as well as the bacterial adhesion and platelet adhesion/activation that occur as follows. A zwitterionic polymer that consists of both cationic and anionic functionalities have been reported as an effective material to achieve these goals, likely resulted from the strongly-adsorbed hydration layer after being immersed in the physiological environment. In this investigation, a novel beta-thiopropionate-based zwitterionic monomer, 2-ammonio-3-((3-(2-hydroxy-3-(methacryloyloxy)propoxy)-3-oxopropyl)thio)-3-methylbutanoate (DPAMA), was synthesized through a facial process. And then the hydrophobic polypropylene was surface modified with this novel zwitterionic polymer through the surface-initiated atom transfer radical polymerization technique. Surface characterization analyses have been employed to investigate the modified surface properties in each reaction stage. In vitro protein adsorption, bacterial adhesion, and platelet compatibility evaluations have shown the polyDPAMA-modified polypropylene surface has significantly reduced fouling characteristics and good hemocompatibility. Henceforth, this novel zwitterionic polyDPAMA grafting PP and the associated grafting reaction scheme have great potential for future clinical applications.

Studies of proliferation and chondrogenic differentiation of rat adipose stem cells using an anti-oxidative polyurethane scaffold combined with cyclic compression culture.

The adipose stem cell is a potential candidate for the autologous chondrocytes repairing approach because of the abundance of fat in the animal body and its versatile differentiation capability. In this study, rat adipose stem cells (rASCs) were seeded into anti-oxidative N-acetylcysteine (NAC) grafted polyurethane (PU) scaffold and then combined with short dynamic compressive stimulation (24 h) to induce rASCs chondrogenesis differentiation in vitro. The inner pore surface of the PU scaffold was first modified via alginate and type I collagen to promote rASCs adherence. The modified layers crosslinked by genipin showed outstanding stability after ultrasonic treatment, indicating the modified layers were stable and can keep the cells adhesion well during dynamic compressive stimulation. After inner pore surface modification and 10 mM NAC grafting, the PU scaffold-A-C-G (graft 10 mM NAC) has shown the best proliferation efficiency with homogeneous cell distribution after 72hr static culture. After short term dynamic compressive stimulation, significant gene expression in chondrogenic markers, Sox-9, and Aggrecan, were noted in both PU scaffold-A-C-G and PU scaffold-A-C-G (graft 10 mM NAC). Considering the cell proliferation efficiency and gene expression, the anti-oxidative NAC grafted PU scaffold combined with short term dynamic compressive stimulation could be useful for cell culturing in stem cell therapy.

The inhibition of TNF-alpha-induced E-selectin expression in endothelial cells via the JNK/NF-kappaB pathways by highly N-acetylated chitooligosaccharides.

Chitooligosaccharides (COS) have been shown to regulate various cellular and biological functions. However, the effect of COS on inflammatory responses of the cells remains unclear. We investigated the regulatory effect of highly N-acetylated COS (NACOS) on tumor necrosis factor-alpha (TNF-alpha)-induced endothelial cell (EC) E-selectin expression, which is crucial for leukocyte recruitment. ECs were kept as controls or pre-treated with NACOS for different times, and then stimulated with TNF-alpha for 4h. The results show that pre-treating ECs with NACOS inhibited the TNF-alpha-induced E-selectin expression in a dose- and time-dependent manner. This NACOS-mediated inhibition in E-selectin expression was regulated at the transcriptional level, but not due to changes in mRNA stability. Stimulation of ECs with TNF-alpha-induced rapid increases in the phosphorylation of their mitogen-activated protein kinases (MAPKs) [extracellular signal-regulated kinase (ERK), c-Jun-NH2-terminal kinase (JNK), and p38 MAPK]; the inhibitor for JNK (i.e., SP600125), but not those for ERK (i.e., PD98059) and p38 MAPK (i.e., SB203580), attenuated this TNF-alpha-induced E-selectin expression. Pre-treating ECs with NACOS inhibited the TNF-alpha-induced JNK activation, suggesting that JNK was involved in the inhibitory effect of NACOS on TNF-alpha-induced E-selectin expression. Pre-treating ECs with NACOS inhibited the TNF-alpha-induced p65 and p50 mRNA expressions. Gel shifting and chromatin immunoprecipitation assays showed that NACOS blocked the TNF-alpha-induced increases in the binding activity and in vivo promoter binding of nuclear factor-kappaB (NF-kappaB) in ECs. Our findings provide a molecular mechanism by which NACOS inhibit TNF-alpha-induced E-selectin expression in ECs, and a basis for using NACOS in pharmaceutical therapy against inflammation.

Surface characterization and platelet adhesion studies for the mixed self-assembled monolayers with amine and carboxylic acid terminated functionalities.

The mixed self-assembled monolayers (SAM) prepared from long chain alkanethiols, HS(CH(2))(11)NH(2) and HS(CH(2))(10)COOH, on gold are employed as the model surface for investigating the interactions between the biological environment and synthetic surface. A distinctive SAM preparation scheme was utilized in this investigation. The triethylamine was added to the alkanethiol solution during SAM formation and then followed by additional rinsing of SAM with 10% CH(3)COOH or 1% HCl ethanolic solution. The contact angle values of NH(2) + COOH mixed SAMs were between those of the pure SAMs, except that it was prepared with solution mole fraction of amine-terminated alkanethiol at 0.2. X-ray photoelectron spectroscopy (XPS) analysis has indicated that these two distinctive SAM preparation procedures had both resulted in a reduction in oxidized sulfur species on pure --NH(2) terminated SAM. However, the procedure utilizing 1% HCl ethanolic washing solution was more effective in reducing the unbound thiol fraction and to form a pure --NH(2) SAM with better quality. XPS analysis has also revealed that the surface of NH(2) + COOH mixed SAMs was "amine-rich". In vitro platelet adhesion assay has shown that the amount of adherent platelets on pure positive charged --NH(2) terminated SAM is less than that on anionic --COOH terminated counterpart in both acidic ethanolic washing schemes. Moreover, the lowest platelet adhesion density was noted on the mixed SAM surfaces with surface amine mole fraction at 0.51 and 0.57. This finding suggests that the surface charge with near neutrality might be of importance in reducing platelet adhesion and activation on artificial biomaterial.

Effects of silane- and MDP-based primers application orders on zirconia-resin adhesion-A ToF-SIMS study.

OBJECTIVE: To evaluate the 3-methacryloyloxypropyltrimethoxysilane (MPS)- and 10-methacryloyloxydecyl-dihydrogen-phosphate (MDP)-base primers, in their single or sequential applications, with regard to modifying zirconia surfaces and improving resin-zirconia adhesion. METHODS: Zirconia disks received different treatments: without primer (Zr), MPS-base primer (S), MDP-base primer (M), MPS/MDP mixture (SMmix), MPS followed by MDP (SM), and MDP followed by MPS (MS). The compositions and chemical interactions of the coatings to zirconia were analyzed using time-of-flight secondary ion mass spectrometry (ToF-SIMS) and reconstructed 3D ion images. Surface wettability of these coatings to water and resin adhesive was assessed. The shear bond strength (SBS) between resin and the treated zirconia was also examined before and after thermocycling. RESULTS: Groups S and MS presented substantial OH- ions in the coatings and zirconia substrate. PO2- and PO3- fragments existed in all MDP-treatment groups with various proportions and distributions, while groups M and SM showed higher proportions of PO3- and the zirconium phosphate related ions. In 3D ion images, PO3- in groups M and SM was denser and segregated to the interface, but was dispersed or overlaid above PO2- in SMmix and MS. All the primers increased the surface wettability to water and resin, with M and SM presenting superhydrophilic surfaces. All MDP-treatment groups showed improved SBS before thermocycling, while M and SM retained higher SBS after this. SIGNIFICANCE: The MDP-base primer shows a relevant function in facilitating POZr bonding and enhancing resin-zirconia bonding. The co-treated MPS impairs the chemical activity of MDP, especially if it is the final coat.

Solvent and concentration effects on the surface characteristics and platelet compatibility of zwitterionic sulfobetaine-terminated self-assembled monolayers.

The structural organization of a monolayer influences biological responses as the material makes contact with the bodily fluid. Zwitterionic materials containing the sulfobetaine functionalities have been shown to exhibit protein-repelling characteristics. In this study, the effect of solvent and thiol concentrations on the sulfobetaine-terminated SAM (self-assembled monolayer) is discussed. Four different types of solvents were selected: deionized water, PBS, methanol and ethanol. The total thiol concentration was set at either 2mM or 0.1 mM. X-ray photoelectron analyses indicated that all SAMs demonstrated similar chemical configurations. Reflection adsorption infrared spectroscopy showed that conformation of the SAMs was more organized when prepared from a 0.1 mM solution compared to a 2 mM solution. The contact angle of the SAMs prepared from 2 mM concentration was dependent upon the solvent utilized and was more hydrophobic than the SAMs prepared from 0.1 mM concentration. Moreover, all of these sulfobetaine-terminated SAMs showed a fairly negative zeta potential in PBS at pH 7.4. After contact with blood, these sulfobetaine-terminated SAMs demonstrated distinct platelet reactivity among each other. The highest platelet compatibility was shown on the SAMs prepared in 0.1 mM solution and the one formed in 2 mM ethanolic solution, where they exhibited a more organized conformation and enhanced hydrophilic properties. These properties might be caused by the different hydration layers, which are affected by the assembly conditions on the topmost monolayer. This study demonstrated that optimizing solvent and concentration conditions could control the structural organization of zwitterionic sulfobetaine-terminated SAMs and, consequently, modify biomedical properties.

Riboflavin-ultraviolet-A-induced collagen cross-linking treatments in improving dentin bonding.

OBJECTIVES: To evaluate the collagen cross-linkers, riboflavin-ultraviolet-A (RF/UVA) and glutaraldehyde, with regard to their efficacy in cross-linking the dentinal collagen and improving dentin bonding. METHODS: Glutaraldehyde and different RF/UVA protocols (0.1%RF/1-minUV, 0.1%RF/2-minUV, and 1%RF/1-minUV) were first evaluated by gel electrophoresis to determine their abilities of collagen cross-linking. The mechanical properties of acid-etched dentin receiving these cross-linking treatments were examined in either dry or wet condition by a nanoindentation test. Fifteen teeth with exposed occlusal dentin received the microtensile bond strength (µTBS) test. The teeth were primed either with RF/UVA or glutaraldehyde, followed by adhesive treatment and composite restorations, and then cut into resin-dentin microbeams. Half of the microbeams received the µTBS test after 24h, and the other half received test after 5000 thermocycles. Nanoleakage at the bond interface was examined under TEM. The alignments of collagen fibrils in the hybrid layers were also defined by an image analysis. RESULTS: Gel electrophoresis showed that glutaraldehyde induced strong collagen gelation, while RF/UVA generated milder collagen cross-linking. Glutaraldehyde, 0.1%RF/2-min-UVA, and 1%RF/1-minUV showed higher stiffness compared to untreated and 0.1%RF/1-minUV in wet condition. All the crosslinking treatments improved early µTBS, but 0.1%RF/2-minUVA treatment maintained high µTBS after theromocycles. Under TEM, glutaraldehyde-treated dentin showed dense and enclosed collagen network on the adhesive interface. 0.1%RF/2-minUVA showed the least nanoleakage, and this could be associated with the suspended collagen fibrils in the hybrid layer. SIGNIFICANCE: 0.1%RF/2-minUVA treatment enhanced resin-dentin bond possibly through enhancing the stiffness and maintaining the expanding collagen matrix in the hybrid layer.

Surface phosphorylation for polyelectrolyte complex of chitosan and its sulfonated derivative: surface analysis, blood compatibility and adipose derived stem cell contact properties.

Many studies have tried to look for the application of chitosan in tissue engineering since its structure is similar to glycoaminoglycans, the main components of the extracellular matrix. Previous studies had indicated that the incorporation of sulfonic or phosphonic functionalities would be beneficial to the growth of certain cells. However, no study has explored the effect of incorporation of both above-mentioned anionic functionalities onto the chitosan structure. In this study, we have surface-phosphorylated the polyelectrolyte film formed by chitosan and water-soluble sulfonated chitosan with the aim to incorporate phosphonic and sulfonic functionalities onto the film surface. Surface analyses by ESCA and ATR-FT-IR have shown that these two functional groups have been successfully grafted onto the surface, and that the ratio of P/S was dependent upon the weight ratio of phosphorylation agents added. Blood compatibility evaluation indicated that phosphorylated polyelectrolyte complexes extended the plasma recalcification time as compared to non-treated chitosan and direct-phosphorylated chitosan film. In addition, these phosphorylated polyelectrolyte complexes showed similar or slightly less platelet reactivity than the non-phosphorylated counterpart. In contrast, significant platelet activation and adhesion were noted on the direct-phosphorylated chitosan. This implicated the incorporation of sulfonic acid onto the phosphorylated surface can increase the platelet compatibility. An adipose-derived stem cell incubation study has demonstrated that the incorporation of both phosphonic and sulfonic acid functionalities onto the chitosan surface can enhance the stem cell growth. Therefore, the phosphorylated polyelectrolyte complexes were not only blood compatible but also stem cell compatible, and could be a novel biomaterial in tissue-engineering applications.

Surface characterization and platelet compatibility evaluation of binary mixed self-assembled monolayers containing novel sulfonic acid terminated alkanethiol.

The interactions between the biological environment and artificial surface are greatly influenced by the surface characteristics of substrate. Self-assembled monolayer prepared by long-chain alkanethiol on gold has been considered as a good model surface to study the effects of surface characteristics upon biological responses. In this study, two different series of mixed SAMs prepared by lab-synthesized sulfonic acid terminated alkanethiol with hydrophobic -CH(3) or hydrophilic -OH terminated one were characterized. It was noted that the surface hydrophilicity of -SO(3)H/-CH(3) mixed SAMs was increased with the solution mole fraction of -SO(3)H terminated thiol. However, the surfaces were all hydrophilic on the -SO(3)H/-OH mixed SAMs. All of these mixed SAMs were "-SO(3)H poor" and negatively charged on surface. In vitro platelet adhesion study indicated that -SO(3)H/-OH mixed SAMs exhibited a better platelet compatibility than -SO(3)H/-CH(3) mixed ones, suggesting a surface with fair hydrophilicity and least negative surface charges might be of potential as a candidate for developing a platelet/blood compatible artificial surface.

Synthesis and characterization of poly(vinyl alcohol) membranes with quaternary ammonium groups for wound dressing.

2-[(acryloyloxy)ethyl]Trimethylammonium chloride (AETMAC) was grafted onto poly(vinyl alcohol) (PVA) using ceric ammonium nitrate (CAN) as a redox initiator. A series of graft co-polymer (PVA-g-PAETMAC) membranes with different contents of AETMAC were prepared with a casting method. The incorporation of AETMAC into PVA chains was confirmed by element analysis and Fourier transform infrared spectroscopy. The effects of grafting on the thermal properties, water take, water vapor transmission rate (WVTR), contact angle, antibacterial activity and cytotoxicity of PVA-g-PAETMAC membranes were investigated. The experiment results showed that PVA-g-PAETMAC membrane has a higher equilibrium swelling ratio, surface hydrophilicity and WVTR than pure PVA membrane. Moreover, the higher the content of AETMAC, the higher were equilibrium swelling ratio, surface hydrophilicity and WVTR. In vitro bacterial adhesion study demonstrated a significantly reduced number of Staphylococcus aureus and Escherichia coli on PVA-g-PAETMAC surfaces when compared to PVA surface. In addition, no significant difference in the in vitro cytotoxicity was observed between PVA and PVA-g-PAETMAC membranes. The presence of quaternary ammonium groups did not reduce L929 cell growth. Therefore, the PVA-g-PAETMAC membranes have the potential for wound-dressing application.

Bioactivity and platelet adhesion study of a human thrombomodulin-immobilized nitinol surface.

Nitinol is a newly developed biomaterial that is gaining popularity in many biomedical applications. It has been reported that nitinol would not induce an inflammatory response and repulsion by the immunization after implantation in the human body. Besides, nitinol is a kind of shape memory alloy, which can memorize shapes at different temperatures. This can improve the convenience in surgery. However, nitinol has poor blood compatibility, so that further modification was needed to improve the antithrombogenicity. Human thrombomodulin (hTM), an endothelial-cell-associated glycoprotein, can be considered as a natural potent anticoagulant by converting thrombin from a procoagulant protease to an anticoagulant. In this study, the surface of nitinol was pre-activated by utilizing silanization with amino-terminated silane. The incorporated amino groups were available for the subsequent covalent immobilization of hTM by 2,4,6-trichloro-1,3,5-triazine (TCT), the coupling reagent. The surface density of immobilized hTM was determined by the Bradford method. The bioactivity of immobilized hTM and blood compatibility of various nitinol substrates were evaluated by the protein C activation assay and platelet adhesion test. It was observed that the immobilized hTM still had the ability to enhance protein C activation, though its activity was lower than the free hTM in solution. Furthermore, the platelet adhesion test showed that only a few platelets were adhered on the hTM-immobilized nitinol substrate. Therefore, the immobilization of thrombomodulin onto nitinol substrate could improve the blood compatibility of nitinol and might have the potential of application in antithrombogenic medical applications.

Surface characterization and in vitro platelet compatibility study of surface sulfonated chitosan membrane with amino group protection-deprotection strategy.

Glycosaminoglycans (GAGs) are the main components of the extracellular matrix (ECM). Studies have indicated that scaffolds modified by GAGs could improve cell proliferation and differentiation. Chitosan, the second-most abundant nature polysaccharide, has a structure similar to that of GAGs. Due to its relatively lower cost as compared to GAGs, many researchers have tried to incorporate sulfonate or carboxyl groups into the chitosan structure with the aim to form a GAG-like structure. However, these modifications were carried out on the reactive amino groups that were thought as the major character, resulting in special biological properties associated with chitosan. Such a decrease of amino-functional group density would very likely alter the specific biological properties of chitosan. Therefore, an amino group protection-deprotection strategy was explored in this study for surface sulfonation of chitosan membrane with the aim to imitate GAG structures. Various surface chemical characterization results, as well as surface zeta potential measurements have indicated that both sulfonate/sulfonic and amino functionalities were coexistent on the deprotected sulfonated chitosan specimen. In vitro platelet adhesion testing has shown that such a deprotected sulfonated chitosan membrane can increase the amount of platelet adhesion while keep those adhered remained unactivated. At the same time the presence of deprotected sulfonated chitosan film extended the plasma recalcification time value. With this protection-deprotection strategy, a further chemical grafting of bioactive molecules, such as RGD peptide, using the recovered amino functionalities, can be pursued on these sulfonated chitosan specimens.