碱性成纤维细胞及角蛋白对丝素蛋白膜改性的实验研究  被引量:1

Characteristics of silk fibroin membrane modified by basic fibroblast growth factor and keratin

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作  者:刁小娟[1] 赵东旭[1] 李珺山 陈帼玲 

机构地区:[1]北京理工大学生命学院,100081 [2]Med Star Georgetown University Hospital

出  处:《中华细胞与干细胞杂志(电子版)》2015年第4期29-32,共4页Chinese Journal of Cell and Stem Cell(Electronic Edition)

基  金:973计划课题(2005CB623906)

摘  要:目的探讨碱性成纤维细胞生长因子(bFGF)及角蛋白对丝素蛋白膜改性后的生物相容性改变。方法还原法制备水溶性人发角蛋白,用共混法使人发角蛋白与丝素蛋白形成共混膜;化学交联法用bFGF对丝素膜进行表面修饰形成交联膜。通过MTT法及荧光显微镜观察检测3T3细胞在三种丝素基材料上的增殖情况。通过接触角测定,对上述三种材料亲水性进行表征。不同的丝素基材料对细胞增殖的促进情况以及接触角的测定2组间SUV_(max)比较采用两样本t检验。结果制备的人发角蛋白分子量范围在44-66KDa之间,条带清晰。MTT法及显微镜观察表明,含角蛋白40﹪共混膜对细胞生长的促进作用最为明显(0.718±0.03,P<0.05),交联膜次之(0.545±0.022,P<0.05),最后是丝素膜(0.463±0.027,P<0.01)。接触角测定结果表明,共混膜的接触角(17.5±1.6,P<0.01)最小,交联膜(47±1.8,P<0.01)次之,丝素膜的接触角(61±1.5,P<0.05)最大,即共混膜黏附性最好。结论制备的40﹪共混膜具有最强生物相容性,b FGF修饰的丝素蛋白交联膜次之,最后是丝素膜。Objective The biocompatibility of silk-based membrane basis which was modified by basic fibroblast growth factor (bFGF) and keratin was evaluated. Methods The water-soluble keratin was made from hair keratin via reduction reaction and blended with silk fibroin. Basic fibroblast growth factor (bFGF) was crosslinked to the surface of silk fibroin membrance. The effects of different silk fibrin-based materials on 3T3 cell proliferation were evaluated using MTT method and cells were observed by fluorescence microscopy. The hydrophilic property was characterized through the measurement of contact angle of the silk based materials. SUVmax was compared using two sample t test. Results Molecular weight of prepared keratin ranged between 44-66 KDa, and the strips were clear. MTT and microscopic observations showed that blended membrane (0.718 土0.03, P 〈 0.05) was most effective in promoting cell growth, followed by erosslinked membrane (0.545 土 0.022, P 〈 0.05), and silk fibroin membrane (0.463 土 0.027, P 〈 0.01). Blended membrane had the smallest contact angle (17.5 土 1.6, P 〈 0.01) followed by the crosslinked membrane (47.5土1.8, P 〈 0.01), and the silk fibroin membrance (61 土 1.5, P 〈 0.05), so the blend membranes has the minimum contactangle. Conclusion The blend membrane had the best biocompatibility, then the silk fibroin cross-linked by bFGF, finally was the silk fibroin membrane.

关 键 词:成纤维细胞 丝素蛋白 生长因子 角蛋白 

分 类 号:R318.08[医药卫生—生物医学工程]

 

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