New biocompatible polypyrrole-based films with good blood compatibility and high electrical conductivity

A novel O-butyryl chitosan (OBCS)-grafted polypyrrole (PPy) film was described. The immobilization was accomplished by photocrosslinking the OBCS onto PPy films under ultraviolet light irradiation. The surfaces of OBCS-grafted PPy film were characterized by attenuated total reflection Fourier transform infrared (ATR-FTIR) spectroscopy and electron spectroscopy for chemical analysis (ESCA). The blood compatibility of the OBCS-grafted PPy film was evaluated by platelet-rich plasma (PRP) contacting experiments and protein adsorption experiments in vitro. These results have demonstrated that the surface with immobilized OBCS shows much less platelet adhesive and fibrinogen adsorption compared to the control surface. The bulk conductivity values of PPy films were measured by a modified four-probe method. The composite films have both good blood compatibility and high electrical conductivity that make them suitable for using as potential biomaterials, such as electrically conducting blood vessel and functionally haemocompatible substrate of biosensor used directly in whole blood.     

Various approaches to modify biomaterial surfaces for improving hemocompatibility

In this paper, the mechanism of thrombus formation on the surface of polymeric materials and the various approaches of modifying biomaterial surfaces to improve their hemocompatibility are reviewed. Moreover, the blood compatibility of the cellulose membrane grafted with O-butyrylchitosan (OBCS) by using a radiation grafting technique was studied. Surface analysis of grafted cellulose membrane was verified by attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR) and electron spectroscopy for chemical analysis (ESCA), which confirmed that OBCS was successfully grafted onto the cellulose membrane surfaces. Blood compatibility of the grafted cellulose membranes was evaluated by platelet rich plasma (PRP) contacting experiments and protein adsorption experiments using blank cellulose membranes as the control. The blood compatibility of OBCS grafted cellulose membranes is better than that of blank cellulose membranes. These results suggest that the photocrosslinkable chitosan developed here has the potential of serving in blood-contacting applications in medical use.     

Introduction of O-butyrylchitosan with a photosensitive hetero-bifunctional crosslinking reagent to silicone rubber film by radiation grafting and its blood compatibility

Based on an in vitro test for an improvement of the blood compatibility of silicone rubber (SR) films by grafting O-butyrylchitosan (OBCS), OBCS was covalently immobilized onto SR film surface using the photosensitive hetero-bifunctional crosslinking reagent, 4-azidobenzoic acid, which was previously bonded to OBCS by reaction between an acid group of the crosslinking reagent and a free amino group of OBCS. Surface properties of SR film were investigated by attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR), electron spectroscopy for chemical analysis (ESCA) and the water contact angle measurements. The blood compatibility of SR film was evaluated by platelet rich plasma (PRP) contacting experiments and the results were observed by scanning electron microscopy (SEM). The state of platelet adhesion was described. The suitable modifications could be carried out to tailor SR films biomaterial to meet the specific needs of different biomedical applications. These results suggest that the blood compatible of SR films/OBCS films show their suitability as potential biomaterials.     

Blood compatibility of polyurethane surface grafted copolymerization with sulfobetaine monomer

Surface modification is an effective way to improve the hemocompatibility and remain bulk properties of biomaterials. Recently, polymer tailed with zwitterions was found having good blood compatibility. In this study, the grafting copolymerization of sulfobetaine onto polyurethane surface was obtained through two steps. In the first step, polyurethane film coupled with vinyl groups was obtained through the reaction between the carboxyl group of acrylic acid (AA) and the NH-urethane group of polyurethane by dicyclohexylcarbodiimide (DCC). In the second step, sulfobetaine was grafted copolymerization on the surface using AIBN as an initiator. The reaction process was monitored with ATR-IR spectra and X-ray photoelectron spectroscopy (XPS) spectra. The wettability of films was investigated by water contact angle measurement. The blood compatibility of the grafted films was evaluated by platelet adhesion in platelet rich plasma (PRP) and protein absorption in bovine fibrinogen (BFG). Low platelet adhesion was observed on the grafted films incubated in PRP for 1 and 3 h, respectively. The protein absorption was reduced on the grafted films after incubated in bovine fibrinogen for 2 h. All of these results revealed that the improved blood compatibility was obtained by grafting copolymerization with zwitterionic monomer of sulfobetaine onto polyurethane film. In addition, introducing vinyl groups onto surface through DCC and AA is a novel method to functionalize polyurethane for further modification.     

Synthesis and characterization of nitric oxide-releasing sol-gel microarrays

Diazeniumdiolate-modified sol-gel microarrays capable of releasing low levels of nitric oxide are reported as a viable means for improving the blood compatibility of a surface without fully modifying the underlying substrate. Several parameters are characterized including: (1) NO surface flux as a function of sol-gel composition and microarray geometry; (2) microstructure dimensions and spacing for optimal blood compatibility; and (3) the effect of sol-gel surface modification on analyte accessibility to platinum electrodes. The sol-gel microarrays release biologically relevant levels of NO under physiological conditions for >24 h. In vitro platelet adhesion assays indicate that a NO surface flux of 2.2 pmol cm(-2) s(-1) effectively reduces platelet adhesion to glass substrates modified with sol-gel microstructures separated by 50 microm. The blood compatibility observed for these micropatterned surfaces is comparable to NO-releasing sol-gel films. When the separation between NO-releasing microstructures is reduced to 10 microm, the NO surface flux required to reduce platelet adhesion is lowered to 0.4 pmol cm(-2) s(-1). Finally, the oxygen response of platinum electrodes modified with NO-releasing sol-gel microarrays indicates that selective modification via micropatterning enhances analyte accessibility to the sensor surface.     

Blood compatibility of surface-engineered poly(ethylene terephthalate) via o-carboxymethylchitosan

Poly(ethylene terephthalate) (PET) films were treated by argon plasma following by graft copolymerization with acrylic acid (AAc). The obtained PET-surface grafted PAA (PET-g-PAA) was coupled with chitosan (CS) and o-carboxymethylchitosan (OCMCS) molecules, respectively. Their surface physicochemical properties were characterized by X-ray photoelectron spectroscopy (XPS), water contact angle and streaming potential measurements. The PET-g-PAA surface containing carboxylic acid, CS immobilized PET surface containing amino and OCMCS immobilized PET surface containing both carboxylic acid and amino groups, make the PET surface exhibited a hydrophilic character. The blood compatibility was evaluated by platelet contacting experiments and protein adsorption experiments in vitro. The results demonstrate that the PET surface coupling OCMCS shows much less platelet adhesive and fibrinogen adsorption compared to the other surface modified PET films. The anticoagulation of PET-OCMCS is ascribed to the suitable balance of hydrophobicity/hydrophilicity, surface zeta potential and the low adsorption of protein.     

Chitosan based surfactant polymers designed to improve blood compatibility on biomaterials

We developed chitosan based surfactant polymers that could be used to modify the surface of existing biomaterials in order to improve their blood compatibility. These polymers consist of a chitosan backbone, PEG side chains to repel non-specific protein adsorption, and hexanal side chains to facilitate adsorption and proper orientation onto a hydrophobic substrate via hydrophobic interactions. Since chitosan is a polycationic polymer, and it is thrombogenic, the surface charge was altered to determine the role of this charge in the hemocompatibility of chitosan. Charge had a notable effect on platelet adhesion. The platelet adhesion was greatest on the positively charged surface, and decreased by almost 50% with the neutralization of this charge. A chitosan surface containing the negatively charged SO(3)(-) exhibited the fewest number of adherent platelets of all surfaces tested. Coagulation activation was not altered by the neutralization of the positive charge, but a marked increase of approximately 5-6 min in the plasma recalcification time (PRT) was displayed with the addition of the negatively charged species. Polyethylene (PE) surfaces were modified with the chitosan surfactant resulting in a significant improvement in blood compatibility, which correlated to the increasing PEG content within the polymer. Adsorption of the chitosan surfactants onto PE resulted in approximately an 85-96% decrease in the number of adherent platelets. The surfactant polymers also reduced surface induced coagulation activation, which was indicated by the PEG density dependent increase in PRTs. These results indicate that surface modification with our chitosan based surfactant polymers successfully improves blood compatibility. Moreover, the inclusion of either negatively charged SO(3)(-) groups or a high density of large water-soluble PEG side chains produces a surface that may be suitable for cardiovascular applications.     

316L钢表面纳米银镀层的制备、性质及血液相容性

采用恒电位电沉积方法在316L不锈钢表面沉积银纳米镀层, 并将钢板置于全氟硅烷溶液中浸泡, 通过动态凝血实验、 抗凝血时间测定、 血小板黏附实验、 溶血实验和蛋白吸附实验等手段, 测试材料的血液相容性. 结果表明, 通过上述方法可明显改善316L不锈钢的血液相容性, 而抗凝血性能、 溶血率及纤维蛋白吸附量不亚于裸钢板. 与316L裸不锈钢相比, 银镀膜全氟硅烷浸泡316L不锈钢具有良好的血液相容性, 是一种比较理想的冠状动脉支架材料, 具有良好的应用前景.     

Surface modification of SPEU films by ozone induced graft copolymerization to improve hemocompatibility

Surface modification of segmented poly(ether urethane) (SPEU) by graft copolymerization with N,N′-dimethyl-N-methacryloyloxyethyl-N-(3-sulfopropyl) ammonium (DMMSA), a zwitterionic sulfobetaine structure, was conducted. A simple two-step procedure for grafting of DMMSA onto the surface of SPEU film was used. The surface was first treated with ozone to introduce active hydroperoxide groups. The active surface was then exposed to the DMMSA solution in the sealed tube. Grafted SPEU film was characterized by ATR–FTIR, XPS and contact angle measurement. ATR–FTIR and XPS investigations confirmed the graft copolymerization. The monomer concentration, copolymerization temperature and time were varied to maximize the efficiency of DMMSA grafting. The equilibrium water content (EWC) and contact angle measurements showed that the hydrophilicity of the film had been greatly improved. The blood compatibility of the grafted films was evaluated by platelet adhesion in platelet rich plasma (PRP), deposits in blood control and protein adsorption in bovine fibrinogen using SPEU film as the control. No platelet adhesion and no thrombus were observed for the grafted films incubated in PRP for 300 min and in blood for 120 min, respectively. The protein adsorption was reduced on the grafted films after incubation in bovine fibrinogen for 120 min. These results proved that improved blood compatibility was obtained by grafting this new zwitterionic sulfobetaine structure monomer onto SPEU film.     

血液相容材料的合成研究Ⅷ.磺铵两性离子单体在聚醚氨酯表面的臭氧活化接枝

生物相容性 ,特别是血液相容性是生物医用材料极其重要的性能[1] .提高不凝血性一直是生物材料研究与发展 (Ｒ Ｄ)的主要内容之一 ,半个多世纪来 ,不凝血材料的Ｒ Ｄ已取得了很大的发展[2 ] .但还不能满足心血管植入物 (Ｃａｒｄｉｏｖａｓｃｕｌａｒｉｍｐｌａｎｔｓ)及心血管医物 (Ｃａｒｄｉｏｖａｓｃｕｌａｒｄｅｖｉｃｅｓ)对不凝血性的需要 .Ｒａｔｎｅｒ[3 ] 在最近一次的血液相容性问题研讨会上再次强调了不凝血材料研究的紧迫性 .会议的报告也反映了该领域的研究现状 ,并提出了今后要研究的问题等 .目前不凝血性较好的材料仅有聚…     

Platelet adhesion and protein adsorption on silicone rubber surface by ozone-induced grafted polymerization with carboxybetaine monomer

Platelet adhesion and protein adsorption on the silicone rubber film grafted with N,N'-dimethyl-N-methacryloyloxyethyl-N-(2-carboxyethyl) ammonium (DMMCA) was studied. The grafting was carried out by means of ozone-induced method and was confirmed by ATR-FTIR and XPS investigations. The grafted films possessed relatively hydrophilic surface revealed by contact angle measurement. The blood compatibility of the grafted film was evaluated in vitro by platelet adhesion in platelet-rich plasma (PRP) and protein absorption in bovine fibrinogen (BFG) using silicone film as the reference. No substantial platelet adhesion was observed for the grafted films incubated in PRP for 60 and 180 min. The protein absorption was also significantly reduced after incubated in bovine fibrinogen for 60 min. Both the results indicated that the blood compatibility of silicone rubber was greatly improved by ozone-induced grafting of carboxybetaine zwitterionic polymer onto its surface.     

Persistent Polarization in Polymers and Blood Compatibility

The use of persistent electrical polarization in polymers has been explored as a means of reducing surface thrombogenicity. Parallel efforts have been carried out on the characterization of surface properties related to the electret state in polymers, on the stability of the electret state in desirable prosthetic materials under simulated blood pumping conditions, and on in vitro and in vivo blood compatibility experiments. Preliminary correlations among critical surface tension, electrical polarization, and blood compatibility have been obtained. In vitro blood compatibility has been explored using both a modified Lee-White clotting test and a platelet adsorption experiment. No significant difference in clotting time was found for polarized samples; however, those with a negative charge did adsorb fewer platelets. A series of in vivo studies has been carried out using electrified right atrial flags (36) of poly-vinylidene chloride and vena cava rings (22) of FEP Teflon, Hypalon rubber, and Hypalon/2% polysulfonate. Negatively polarized samples showed improved blood compatibility in all cases. About 2/3 of the negatively charged atrial flags showed little or no thrombus and the remainder moderate thrombus whereas 2/3 of the positively charged flags showed severe thrombus. About 3/4 of the negative ring implants showed little or no thrombus. A correlation with anomalous contact angle or low charge was found in 4 of 6 cases of the rings with moderate to heavy thrombus.     

Platelet adhesive resistance of polyurethane surface grafted with zwitterions of sulfobetaine

A possible approach to improve the blood compatibility of poly(etherurethane)s (PU) involves the covalent attachment of key molecular on its surface. Recently, polymer tailed with zwitterions was found having good blood compatibility. The purpose of present study was to design and synthesis a novel nonthrombogenic biomaterial by modifying the surface of poly(etherurethane) with zwitterions of sulfobetaine via HDI spacer. The films of polyurethane were grafted with sulfobetaine by a three-step procedure. In the first step, the film surfaces were treated with hexamethylene diisocyanate (HDI) in toluene at 50 degrees C in the presence of di-n-butyl tin dilaurate(DBTDL) as a catalyst. The extent of the reaction was measured by ATR-IR spectra; a maximum number of free NCO group was obtained after a reaction time of 2.5 h. In the second step, the primary amine group of N,N-diethylethylenediamine (DEA) or N,N-dimethylethylenediamine (DMA) was allowed to react in toluene with isocyanate groups bound on surface. In the third step, two kinds of sulfobetaines were formed in the surface through the ring-opening reaction between tertiary amine of DMA or DEA and 1,3- propanesultone (PS). The reaction process was monitored with ATR-IR spectra and XPS spectra. The wettability of films was investigated by water contact angle measurement. A platelet adhesion experiment was conducted as a preliminary test to confirm the improved blood compatibility of PU. The number of platelets adhering to PU decreased greatly compared to original after 1 h and 3 h of contact with human plate-rich plasma.     

Improved blood compatibility of rapamycin-eluting stent by incorporating curcumin

This paper dealt with improving the blood compatibility of the rapamycin-eluting stent by incorporating curcumin. The rapamycin- and rapamycin/curcumin-loaded PLGA (poly(d,l-lactic acid-co-glycolic acid)) coatings were fabricated onto the surface of the stainless steel stents using an ultrasonic atomization spray method. The structure of the coating films was characterized by Fourier transform infrared spectroscopy (FTIR). The optical microscopy and scanning electron microscopy (SEM) images of the drug-eluting stents indicated that the surface of all drug-eluting stents was very smooth and uniform, and there were not webbings and "bridges" between struts. There were not any cracks and delaminations on stent surface after expanded by the angioplasty balloon. The in vitro platelet adhesion and activation were investigated by static platelet adhesion test and GMP140 (P-selection), respectively. The clotting time was examined by activated partially prothromplastin time (APTT) test. The fibrinogen adsorption on the drug-loaded PLGA films was evaluated by enzyme-linked immunosorbent assay (ELISA). All obtained data showed that incorporating curcumin in rapamycin-loaded PLGA coating can significantly decrease platelet adhesion and activation, prolong APTT clotting time as well as decrease the fibrinogen adsorption. All results indicated that incorporating curcumin in rapamycin-eluting coating obviously improve the blood compatibility of rapamycin-eluting stents. It was suggested that it may be possible to develop a drug-eluting stent which had the characteristics of not only good anti-proliferation but also improved anticoagulation.     

Determination of platelet monoamine oxidase activity by high-performance liquid chromatography with electrochemical detection.

A procedure for determining human platelet monoamine oxidase (MAO) with dopamine (DA) as substrate is described. High-performance liquid chromatography (HPLC) with electrochemical detection (ED) was used to separate and detect components of the reaction mixture. The method for platelet preparation was also improved and only 2 ml of blood were required. Following a 10-min incubation of the platelet preparation with DA in 0.1 M Tris buffer (pH 9.0), excess DA substrate was removed by adsorption on a cation-exchange resin. The reaction product, 3,4-dihydroxyphenylacetaldehyde, was adsorbed on acid-washed alumina, eluted with 0.1 M perchloric acid and analyzed by HPLC. Simple, clean chromatograms were obtained with good reproducibility using 3,4-dihydroxybenzylamine as an internal standard. The within-sample, between-samples and between-day relative standard deviations were 0.9, 3.7 and 6.1%, respectively. The apparent Michaelis constant and maximum velocity were 0.10 mM and 0.37 nmol/min.mg protein, respectively. This HPLC-ED method offers a good alternative to methods using radioactivity.     

Surface-localized release of nitric oxide via sol-gel chemistry

The release of nitric oxide (NO) from polymers has proven to be highly effective at inhibiting platelet adhesion and thus enhancing the blood compatibility of medical implants. Micropatterning techniques were used to design surfaces that release NO while preserving the underlying substrate for other applications (e.g., sensors). Micropatterned NO-releasing substrates based on aminosilane-containing methyltrimethoxysilane sol-gels were prepared and characterized in terms of stability, NO surface flux, and resistance to in vitro platelet adhesion. We have found that surface-localized NO release from substrates modified with sol-gel micropatterns exhibit enhanced blood compatibility relative to controls.     

Surface modification of polyacrylonitrile-based membranes by chemical reactions to generate phospholipid moieties

A novel approach for the surface modification of poly(acrylonitrile-co-2-hydroxyethyl methacrylate) (PANCHEMA) membranes by introducing phospholipid moieties is presented, which involved the reaction of the hydroxyl groups on the membrane surface with 2-chloro-2-oxo-1,3,2-dioxaphospholane (COP) followed by the ring-opening reaction of COP with trimethylamine. The chemical changes of phospholipid-modified acrylonitrile-based copolymers (PMANCP) membranes were characterized by Fourier transfer infrared spectroscopy and X-ray photoelectron spectroscopy. The surface properties of PMANCP membranes were evaluated by pure water contact angle, protein adsorption, and platelet adhesion measurements. Pure water contact angles measured by the sessile drop method on PMANCP membranes were obviously lower than those measured on the PANCHEMA membranes and decreased with the increase of the content of phospholipid moieties on the membrane surface. It was found that the bovine serum albumin adsorption and platelet adhesion were suppressed significantly with the introduction of phospholipid moieties on the membranes surface. These results demonstrated that the described process was an efficient way to improve the surface biocompatibility for the acrylonitrile-based copolymer membrane.     

Immobilization of bovine serum albumin onto porous polyethylene membranes using strongly attached polydopamine as a spacer

Based on the self-polymerization and strong adhesion characteristics of dopamine in aqueous solution, a novel and convenient approach was developed to immobilize protein onto porous polyethylene (PE) membranes. A thin polydopamine (pDA) layer was formed and tightly coated onto PE membrane by dipping simply the membrane into dopamine aqueous solution for a period of time. Subsequently, bovine serum albumin (BSA) was bound onto the obtained PE/pDA composite membranes via the coupling between BSA and the reactive polydopamine layer. The firm immobilization of polydopamine layer and BSA was verified by attenuated total reflection-Fourier transform infrared spectroscopy (ATR-FTIR) and X-ray photoelectron spectroscopy (XPS). The results of water contact angle measurement showed that the hydrophilicity of PE membrane was significantly improved after coating polydopamine and binding BSA. The experiments of blood platelet adhesion indicated that BSA-immobilized PE membrane had better blood compatibility than the unmodified PE and the PE/pDA composite membranes. The investigations on hepatocyte cultures and cell viability revealed that the polydopamine coating endowed PE membrane with significantly improved cell compatibility. Compared to BSA surface, polydopamine surface is more favorable for cell adhesion, growth, and proliferation.     

Blood compatibility and permeability of heparin-modified polysulfone as potential membrane for simultaneous hemodialysis and LDL removal

Heparin was covalently immobilized on PSf membranes to obtain a dialysis membrane with high affinity for LDL. WCA and streaming potential measurements were performed to investigate wettability and surface charge of the membranes. The morphology of the membranes was investigated by SEM. An ELISA was used to measure the adsorption and desorption of LDL on plain and modified PSf. Blood compatibility was studied by measurement of thrombin time, partial thromboplastin time, kallikrein activity and platelet adhesion. It was found that the blood compatibility of the membrane was improved by covalent immobilization of heparin at its surface. However, PSf-Hep membrane showed higher flux recovery after BSA solution filtration, which revealed antifouling property of PSf-Hep membranes.     

Porous and electrically conductive polypyrrole-poly(vinyl alcohol) composite and its applications as a biomaterial

Bulk modification of polypyrrole (PPY) with poly(vinyl alcohol) (PVA) was carried out by the electropolymerization of pyrrole in the presence of PVA in the reaction solution, with tetraethylammonium perchlorate (TEAP) as the electrolyte. The surface morphology of the as-synthesized PPY-TEAP-PVA film was investigated using scanning electron microscopy, and the film was further characterized using X-ray photoelectron spectroscopy, electrical conductivity, the water contact angle, and BET surface area measurements. The PPY-TEAP-PVA composite is electrically conductive, hydrophilic, and microporous with a high surface area. Its potential as a biomaterial was investigated with respect to its blood compatibility and function as a substrate for biosensor fabrication and cell culture. The presence of PVA in the film attenuates blood protein adsorption, and the porous nature of the PPY-TEAP-PVA film results in a 10-fold increase in the amount of glucose oxidase covalently immobilized on the film over that on a nonporous PPY film. PC12 cell attachment and growth on the PPY-TEAP-PVA film was also shown to be enhanced compared with that on tissue culture polystyrene. The attached cells proliferated and formed a monolayer on the film surface after 48 h of seeding.