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1.
The effect of surface chemistry on proliferation and morphology of bone cells cultured on surface modified poly(3‐hydroxybutrate‐co‐3‐hydroxyvalerate) (PHBV) and untreated PHBV was evaluated. The surface of cast PHBV film was physically and chemically immobilized with collagen. For preparing chemically immobilized collagen surface, PHBV film was ozone treated followed by grafting of PMAA chains and the immobilization of collagen. The surface roughness and hydrophilicity of PHBV film were determined by atomic force microscopy (AFM) and contact angle measurements, respectively. It was found that the duration of ozone exposure and monomer concentration used for grafting PMMA chains influenced the amount of collagen immobilized. The cell proliferation on PHBV surfaces with chemically and physically immobilized collagen was compared with untreated PHBV using 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyl tetrazolium bromide (MTT) assay. The bone cell activity on chemically and physically immobilized collagen PHBV films was found to be 246 and 107% for UMR‐106 and 68 and 9% for MC3T3 cell lines, respectively. Although the results are very preliminary, the chemically grafted collagen on PHBV surface provided a favorable matrix for cell proliferation. © 2004 Wiley Periodicals, Inc. J Appl Polym Sci 93: 2445–2453, 2004  相似文献   

2.
Porous poly(3‐hydroxybutrate‐co‐3‐hydroxyvalerate) (PHBV) film was prepared by solute leaching of salt/PHBV cast film. The surface chemistry of the PHBV membrane was modified by performing graft polymerization of methacrylic acid (MAA) on ozone treated porous PHBV film, followed by immobilization of type I collagen. The surface characteristics of the modified and nonmodified porous films were measured by water contact angle. The rat osteosarcoma cell line UMR‐106 osteoblast like cells were used as model cells to evaluate the cell viability on surfaces. The initial cell attachment, growth pattern, and proliferation as measured by MTT assay were used to evaluate the bone cell viability on the modified and nonmodified films. Among the PHBV films studied, the nonmodified porous PHBV and the porous PHBV film type I collagen dip coated showed no significant difference in cell attachment and proliferation, while the porous PHBV membrane that was collagen immobilized after MAA grafting showed considerable activity of osteoblast like cells. © 2005 Wiley Periodicals, Inc. J Appl Polym Sci 98: 1916–1921, 2005  相似文献   

3.
Cell and protein-repellent poly(ethylene glycol) (PEG) hydrogels surfaces were modified to covalently attach cell adhesion proteins and subsequently promote cell adhesion. Collagen was chosen as a cell adhesion protein and covalently immobilized to the hydrogel surface via a 5-azidonitrobenzoyloxy N-hydroxysuccinimide (NHS), bifunctional linker, which has a phenyl azide group and a protein-binding NHS group on either end. Using the photochemistry of phenyl azide groups, the bifunctional linker was chemically fixed to the hydrogel surface by UV exposure and the N-hydroxysuccinimide groups were allowed to react with the free amine groups of collagen. The immobilization of collagen on the PEG hydrogel surface was demonstrated with XPS by confirming the formation of a new nitrogen peak and the resulting amount of immobilized collagen was dependent on the concentration of bifunctional linker. Cell adhesion studies revealed that collagen immobilization resulted in a significant improvement of cell adhesion and spreading on the PEG hydrogel substrates. Photochemical fixation combined with photolithography produced well-defined collagen micropatterns on the PEG hydrogels and cells adhered only on the collagen-modified region due to the lack of adhesion for proteins and cells to PEG.  相似文献   

4.
BACKGROUND: Dextran, a bacterial polysaccharide, has been reported to be as good as poly(ethylene glycol) in its protein‐rejecting and cell‐repelling abilities. In addition, the multivalent nature of dextran is advantageous for surface grafting of biologically active molecules. We report here a method to photochemically bind dextran hydrogel films to aminated poly(ethylene terephthalate) (PET) surfaces in aqueous media using a heterobifunctional crosslinker, 4‐azidobenzoic acid. In order to achieve this, dextran was first functionalized with the crosslinker using carbodiimide chemistry followed by photo‐crosslinking and immobilization onto the nucleophile‐rich aminated PET surfaces. RESULTS: The presence of the immobilized dextran on PET was verified by attenuated total‐reflection Fourier transform infrared spectroscopy, X‐ray photoelectron spectroscopy, scanning electron microscopy and contact angle measurements. The grafted surface was highly hydrophilic due to the heavily hydrated polysaccharide network on the surface as demonstrated by the near zero water contact angle. CONCLUSION: A photochemical method for surface immobilization of dextran onto aminated PET using aryl azide chemistry is a facile technique to generate highly hydrophilic and more hemocompatible surfaces. The aryl nitrenes generated by photolysis produce a metastable, electron‐deficient intermediate, azacycloheptatetraene, which is believed to be responsible for the simultaneous crosslinking of dextran and its immobilization onto the aminated PET surface. The aryl azide chemistry reported here for dextran could be useful as a versatile technique for surface modification of other nucleophile‐rich polymers to create dextran‐ or similar polysaccharide‐immobilized surfaces. Copyright © 2007 Society of Chemical Industry  相似文献   

5.
The DNA adsorption properties of poly‐L ‐lysine‐immobilized poly(2‐hydroxyethyl methacrylate) (pHEMA) membrane were investigated. The pHEMA membrane was prepared by UV‐initiated photopolymerization and activated with epichlorohydrin. Poly‐L ‐lysine was then immobilized on the activated pHEMA membrane by covalent bonding, via a direct chemical reaction between the amino group of poly‐L ‐lysine and the epoxy group of pHEMA. The poly‐L ‐lysine content of the membrane was determined as 1537 mg m?2. The poly‐L ‐lysine‐immobilized membrane was utilized as an adsorbent in DNA adsorption experiments. The maximum adsorption of DNA on the poly‐L ‐lysine‐immobilized pHEMA membrane was observed at 4 °C from phosphate‐buffered salt solution (pH 7.4, 0.1 M; NaCl 0.5 M) containing different amounts of DNA. The non‐specific adsorption of DNA on the plain pHEMA membrane was low (about 263 mg m?2). Higher DNA adsorption values (up to 5849 mg m?2) were obtained in which the poly‐L ‐lysine‐immobilized pHEMA membrane was used. Copyright © 2003 Society of Chemical Industry  相似文献   

6.
Surface wettability on anchorage‐dependent cells has an important role in cell growth rate. In our previous studies, we prepared a wettability gradient on polyethylene (PE) surfaces using corona discharge treatment from a knife‐type electrode whose power increased gradually along the sample length. The PE surfaces were oxidized gradually with increasing power and characterized by Fourier transform infrared spectroscopy, contact angle goniometry, and electron spectroscopy for chemical analysis. The purpose of this study is to determine the rate of proliferation on polymer surfaces with different wettability. The behavior of cell growth for NIH/3T3 fibroblast cells attached on the polymer surfaces with different hydrophilicity was investigated using wettability gradient PE surfaces prepared by corona discharge treatment. They were investigated for the number of grown cells from 24 to 60 h in terms of surface wettability. From the slope of cell number on PE gradient surface versus culture time, the proliferation rates (number of cell/cm2 · h) were calculated. It was observed that the proliferation rate was increased more on positions with moderate hydrophilicity of the wettability gradient surface than on the more hydrophobic or hydrophilic positions, i.e., 1111 (number of cell/cm2 · h) of 57° of water contact angle at the 2.5‐cm position (P < 0.05). This result seems closely related to the serum protein adsorption on the surface: the serum proteins were also adsorbed more on the moderately hydrophilic surface. In conclusion, surface wettability plays an important role in cell adhesion, spreading, and proliferation on the polymer surfaces. © 2004 Wiley Periodicals, Inc. J Appl Polym Sci 92: 599–606, 2004  相似文献   

7.
A novel and effective protocol for the preparation of superparamagnetic immunomicrospheres has been developed. First, micro‐size magnetic poly (methacrylate‐divinylbenzene) (PMA‐DVB) spheres were prepared by a modified suspension polymerization method. The oleic acid coated magnetite (Fe3O4) nanoparticles made by coprecipitation were mixed with monomers of MA, DVB, and initiator benzoyl peroxide (BPO) to form oil in water emulsion droplets with the presence of poly (vinyl alcohol) (PVA‐1788) as a stabilizer. The polymerization reaction was carried out in a 2‐L beaker equipped with four vertical stainless steel baffleplates by increasing the temperature of the mixture at a controlled rate. The resulting magnetic microspheres are micro‐sized (less than 8μm in diameter) and 80 percent of them are in the size ranging from 1 to 5 μm. Then, they were highly functionalized via ammonolysis reaction with ethylenediamine, and the surface amino‐modified magnetic microspheres were obtained. The morphology and properties of these magnetic microspheres were examined by SEM, TEM, VSM, and FT‐IR. Affinity ligand protein A (ProtA) was covalently immobilized to the amino‐modified magnetic microspheres by the glutaraldehyde method. These ProtA‐immobilized magnetic immunomicrospheres were effective for affinity bioseparation processes, as was demonstrated by the efficient immunoaffinity purification of antibodies IgG2a (22mg per gram of microspheres) from mouse ascites. © 2004 Wiley Periodicals, Inc. J Appl Polym Sci 94: 2205–2211, 2004  相似文献   

8.
The enzyme microreactor has considerable potential for use in biotechnological syntheses and analytical studies. Simplifying the procedure of enzyme immobilization in a microreactor is attractive, and it is achievable by utilizing enzyme immobilization techniques and taking advantage of the characteristics of microfluidics. We previously developed a facile and inexpensive preparation method for an enzyme‐immobilized microreactor. The immobilization of enzymes can be achieved by the formation of an enzyme‐polymeric membrane on the inner wall of the microchannel through cross‐linking polymerization in a laminar flow. However, this method is unsuitable for use in conjunction with electronegative enzymes. Therefore, a novel preparation method using poly‐L ‐lysine [poly(Lys)] as a booster and an adjunct for the effective polymerization of electronegative enzymes was developed in this study. Using aminoacylase as a model for an electronegative enzyme, the reaction conditions for the enzyme‐cross‐linked aggregation were optimized. On the basis of the determined conditions, an acylase‐immobilized tubing microreactor was successfully prepared by cross‐linking polymerization in a concentric laminar flow. The resulting microreactor showed a higher stability against heat and organic solvents compared to those of the free enzyme. The developed method using poly(Lys) was applicable to various enzymes with low isoelectric points, suggesting that this microreactor preparation utilizing a cross‐linked enzyme in a laminar flow could be expanded to microreactors in which a broad range of functional proteins are employed.  相似文献   

9.
The epoxy‐group‐containing microspheres from cross‐linked glycidyl methacrylate and methyl methacrylate, poly(GMA–MMA), were prepared by suspension polymerisation. The epoxy groups of the poly(GMA–MMA) microspheres were used for grafting with an anionic polymer polyethylenimine (PEI) to prepare non‐specific affinity adsorbents (poly(GMA–MMA)–PEI) for bilirubin removal. The specificity of the poly(GMA–MMA)–PEI adsorbent to bilirubin was further increased by immobilization of human serum albumin (HSA) via adsorption onto PEI‐grafted poly(GMA–MMA) adsorbent. Various amounts of HSA were immobilized on the poly(GMA–MMA)–PEI adsorbent by changing the medium pH and initial HSA concentration. The maximum HSA content was obtained at 68.3 mg g?1 microspheres. The effects of pH, ionic strength, temperature and initial bilirubin concentration on the adsorption capacity of both adsorbents were investigated in a batch system. Separation of bilirubin from human serum was also investigated in a continuous‐flow system. The bilirubin adsorption on the poly(GMA–MMA)–PEI and poly(GMA–MMA)–PEI–HSA was not well described by the Langmuir model, but obeyed the Freundlich isotherm model. The poly(GMA–MMA)–PEI affinity microspheres are stable when subjected to sanitization with sodium hydroxide after repeated adsorption–desorption cycles. Copyright © 2004 Society of Chemical Industry  相似文献   

10.
Well‐defined and characterized polymeric matrices showing close chemical similarities but wide differences in water uptake and swellability in aqueous medium were used for lipase immobilization. Biphasic networks of 2‐hydroxypropylcellulose (HPC) were synthesized with acrylamide (AAm), methacrylamide (MAAm), N‐isopropylacrylamide (N‐i‐PAAm), and 2‐acrylamido‐2‐methyl‐1‐propanesulfonic acid (AMPSA) and simultaneously crosslinked with N,N‐methylene bisacrylamide in aqueous medium by using simultaneous γ‐radiation technique. Lipase enzyme was produced from a mesophilic bacterial isolate (HBK‐8) and was immobilized onto all the matrices by adsorption method. The activity of the immobilized enzyme was optimized for pH, temperature, and amount of crude enzyme and effect of dehydration. High relative activity for the immobilized enzymes was observed and loss of activity with time was minimal; reusability was found to be good. The activity of the immobilized enzyme was also observed to be good in both esterification and hydrolysis of esters. In the present study, lipase immobilization, hydrolysis of p‐nitrophenyl palmitate, and optimum pH and temperature for substrate hydrolysis were evaluated for different matrices to study polymer structure and enzyme activity relationship in diverse physical environments. © 2004 Wiley Periodicals, Inc. J Appl Polym Sci 92: 3135–3143, 2004  相似文献   

11.
The aim of this study was the immobilization of COOH‐modified poly(ethylene glycol) (PEG) layers onto β‐cyclodextrin‐coated surfaces by formation of inclusion complexes, in view of biosensors applications. To this end, PEGs with one phenyladamantyl and one carboxylic end group (Ad‐PEG‐COOH) were prepared according to a three‐step procedure. After modification of PEG with 4‐toluenesulfonylchloride, the reaction of the tosyl intermediate with the alcoholate of 4‐(1‐adamantyl)‐phenol was carried out in tetrahydrofuran to avoid the formation of by‐products. Then, it was shown by high performance liquid chromatography that the association between β‐cyclodextrin cavities and Ad‐PEG‐COOH polymers was not hindered by the presence of the COOH group. Last, the Ad‐PEG‐COOH polymer was immobilized onto β‐cyclodextrin‐coated gold surfaces by formation of inclusion complexes. The immobilization was performed in water, at room temperature, with a rapid kinetics. After activation of COOH groups with N‐hydroxysuccinimide, β‐lactoglobulin was coupled to the biocompatible PEG layer. Functionalization of the gold surface with β‐cyclodextrin cavities, immobilization of Ad‐PEG‐COOH onto the surface, and coupling of the protein to the reactive PEG layer were followed in real time by surface plasmon resonance imaging system. © 2006 Wiley Periodicals, Inc. J Appl Polym Sci 100: 2362–2370, 2006  相似文献   

12.
Fe3O4/poly(styrene‐co‐maleic anhydride) core–shell composite microspheres, suitable for binding enzymes, were prepared using magnetite particles as seeds by copolymerization of styrene and maleic anhydride. The magnetite particles were encapsulated by polyethylene glycol, which improved the affinity between the magnetite particles and the monomers, thus showing that the size of the microspheres, the amount of the surface anhydrides, and the magnetite content in the composite are highly dependent on magnetite particles, comonomer ratio, and dispersion medium used in the polymerization. The composite microspheres, having 0.08–0.8 μm diameter and containing 100–800 μg magnetite/g microspheres and 0–18 mmol surface‐anhydride groups/g microsphere, were obtained. Free α‐amylase was immobilized on the microspheres containing reactive surface‐anhydride groups by covalent binding. The effects of immobilization on the properties of the immobilized α‐amylase [magnetic immobilized enzyme (MIE)] were studied. The activity of MIE and protein binding capacity reached 113,800 U and 544.3 mg/g dry microspheres, respectively. The activity recovery was 47.2%. The MIE had higher optimum temperature and pH compared with those of free α‐amylase and showed excellent thermal, storage, pH, and operational stability. Furthermore, it can be easily separated in a magnetic field and reused repeatedly. © 2004 Wiley Periodicals, Inc. J Appl Polym Sci 95: 328–335, 2005  相似文献   

13.
We developed a synthetic concept for the immobilization of enzymes onto monolithic supports. In addition, we elaborate on a generally applicable method for the rapid screening of the activity of such immobilized enzymes. For these purposes, we prepared monolithic acrylate‐based systems by electron‐beam (EB)‐triggered free‐radical polymerization within the confines of 200‐μm capillary columns. Aiming for protein immobilization, we subjected the polyacrylate‐based monoliths to EB‐mediated grafting processes to introduce functional surface‐located groups suited for the subsequent generation of functional units that themselves could bind different proteins. For the generation of the functional units, we used ring‐opening metathesis polymerization and free‐radical polymerization. The produced systems were tested for their ability to bind or repel proteins as exemplified by the use of the serine protease trypsin, which was used to catalyze the hydrolysis of N‐α‐benzoyl‐L ‐arginine ethyl ester (BAEE). Finally, the monolith‐immobilized trypsin systems were used for enzymatic peptide synthesis purposes, such as the acyl transfer of BAEE to amino acid amides. Complementarily, we used an immobilized trypsin variant, which we additionally subjected to on‐column chemical modification with succinic anhydride to alter its synthetic properties. © 2010 Wiley Periodicals, Inc. J Appl Polym Sci, 2011  相似文献   

14.
Epoxy group‐containing poly(hydroxyethyl methacrylate/glycidyl methacrylate), p(HEMA/GMA), membrane was prepared by UV initiated photopolymerization. The membrane was grafted with chitosan (CH) and some of them were chelated with Fe(III) ions. The CH grafted, p(HEMA/GMA), and Fe(III) ions incorporated p(HEMA/GMA)‐CH‐Fe(III) membranes were used for glucose oxidase (GOD) immobilization via adsorption. The maximum enzyme immobilization capacity of the p(HEMA/GMA)‐CH and p(HEMA/GMA)‐CH‐Fe(III) membranes were 0.89 and 1.36 mg/mL, respectively. The optimal pH value for the immobilized GOD preparations is found to have shifted 0.5 units to more acidic pH 5.0. Optimum temperature for both immobilized preparations was 10°C higher than that of the free enzyme and was significantly broader at higher temperatures. The apparent Km values were found to be 6.9 and 5.8 mM for the adsorbed GOD on p(HEMA/GMA)‐CH and p(HEMA/GMA)‐CH‐Fe(III) membranes, respectively. In addition, all the membranes surfaces were characterized by contact angle measurements. © 2006 Wiley Periodicals, Inc. J Appl Polym Sci 103: 3084–3093, 2007  相似文献   

15.
陈佳璐  陈国  赵珺  陈宏文 《化工学报》2012,63(4):1175-1181
利用共沉淀法结合高锰酸钾氧化制备所得表面羧基修饰的超顺磁性纳米粒子吸附于罗伊氏乳酸杆菌表面,在磁场协助下实现细胞的固定化。吸附机理分析表明小尺寸相互作用和静电相互作用是磁性纳米粒子与细胞之间的主要作用。分别考察了菌体/磁性粒子质量比、pH、温度、时间等对固定化罗伊氏乳酸杆菌的影响,确定最佳固定条件为菌体与磁性纳米粒子相对质量比为2.25,在pH=3、温度25℃的条件下固定化0.5 h,可实现91%的细胞固定化。最后,对固定化后的细胞进行再培养,与游离细胞相比,两者表现出类似的代谢特征,证实细胞经固定化后仍具有活性。因此,羧基修饰的超顺磁性纳米粒子可成功用于细胞固定化,在不影响细胞活性的情况下,通过磁分离实现细胞的重复利用。  相似文献   

16.
In this article, urease was immobilized in a conducting network via complexation of poly(1‐vinyl imidazole) (PVI) with poly(2‐acrylamido‐2‐methyl‐1‐propanesulfonic acid) (PAMPS). The preparation method for the polymer network was adjusted by using Fourier transform infrared (FTIR) spectroscopy. A scanning electron microscope (SEM) study revealed that enzyme immobilization had a strong effect on film morphology. The proton conductivity of the PVI/PAMPS network was measured via impedance spectroscopy, under humidified conditions. The basic characteristics (Michealis‐Menten constants, pHopt, pHstability, Topt, Tstability, reusability, and storage stability) of the immobilized urease were determined. The obtained results showed that the PAA/PVI polymer network was suitable for enzyme immobilization. © 2010 Wiley Periodicals, Inc. J Appl Polym Sci, 2011  相似文献   

17.
Ai-Fu Che  Xiao-Dan Huang  Ke Yao 《Polymer》2005,46(24):11060-11065
Asymmetric membranes fabricated from poly(acrylonitrile-co-maleic acid) (PANCMA) were immobilized with heparin and/or insulin to improve their surface properties. These biomacromolecule-immobilized PANCMA membranes were prepared by the amination of the membrane surface with ethylenediamine, followed by the reaction of the amino groups with heparin and/or insulin in the presence of 1-ethyl-3-(3-dimethyl amidopropyl) carbodiimide. The surface-modified membranes were analyzed by X-ray photoelectron spectroscopy to confirm the immobilization of the biomacromolecules. Morphological changes on the membrane surface and in the cross section were characterized by scanning electron microscopy. The surface hydrophilicity and hemocompatibility of the studied membranes were evaluated on the basis of water contact angle, platelets adhesion and cell attachment measurements. It was found that, after the immobilization of the biomacromolecules, the water contact angle and the amount of adhered platelets and macrophages on the membrane decreased significantly when compared with the nascent ones, indicating the improvement of surface hydrophilicity. Furthermore, the heparin immobilized membrane showed the best hemocompatibility among the corresponding membranes studied.  相似文献   

18.
Nitrogen plasma was used to amino‐functionalize polystyrene surfaces, which were further modified via the selective introduction of polyamines suitable for the immobilization of biological compounds. This chemical modification was carried out using a multifunctional amine compound linked to glutaraldehyde, leading to the formation of hyperbranched structures at the surface. Up to three generations of branched polymers at the polystyrene (PS) surface were created by successive addition of the functional compounds. Amine functions introduced at the surface were labeled with 2,3,4,5,6‐pentafluorobenzaldehyde and analyzed by X‐ray photoelectron spectroscopy (XPS), confirming the successful attachment of each generation of branching. Finally, bovine serum albumin and trypsin were immobilized on N2‐plasma‐treated PS modified with different amounts of branched graft polymer and found to remain bioactive after immobilization. © 2009 Wiley Periodicals, Inc. J Appl Polym Sci, 2009  相似文献   

19.
Styrene/divinyl benzene‐based macroporous polyHIPE composites were prepared from water‐in‐oil (w/o) high internal phase emulsion (HIPE) templates by using both organo‐modified montmorillonite (MMT) and a nonionic surfactant. For this purpose, Spirulina (Sp) microalgae was immobilized onto Na‐MMT clay by using two different modification techniques. They are based on conventional adsorption in solution (SOL) and novel cryoscopic expansion (C‐XP) assisted adsorption. Highly porous nanocomposites were prepared by using different percentages of modified nanoclays (SpSOLM/SpXPM) with a constant internal phase volume of 80%. The emulsion stability, morphology, and dye adsorption capacities were discussed by paying attention to nanoclay immobilization techniques, clay loading degree and surfactant concentration. The critical amount of nonionic surfactant for formation of the stable neat HIPE template was found to be only 5 vol% with respect to volume of organic phase. However, this amount was further reduced to much less value (2 vol%) with Sp immobilized nanoclays via help of cooperative interactions of Sp and MMT nanoclay. The C‐XP assisted modification of clay led to nanocomposites with 580% higher adsorption capacity for cationic dye. This remarkable benefit was obtained with even 0.5% clay loading and only 2% surfactant concentration. POLYM. ENG. SCI., 58:1229–1240, 2018. © 2017 Society of Plastics Engineers  相似文献   

20.
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