以再生丝素蛋白为原料，采用旋转干燥法，通过控制温度和湿度及乙醇改性，最终得到不溶于水的丝素膜。为了评价再生丝素膜的生物安全性，进行体外细胞毒性、全身过敏反应、迟发型超敏反应、急性毒性反应、热原反应以及溶血性试验。结果：用丝素膜浸提液对体外培养的成纤维细胞进行处理后，细胞平均相对增殖率为92.3%，属于细胞毒性I级。小鼠经由100%丝素膜浸提液多次致敏后再次激发，未产生全身性过敏反应；供试小鼠静脉大剂量注射100%丝素膜浸提液后全部存活，无急性全身毒性反应，7d观察期内小鼠的体质量逐步增长；用100%丝素膜浸提液对家兔进行静脉注射后，兔的单只体温升高幅度为0.2～0.5 ℃，未超0.6 ℃上限，总和也未超1.4 ℃上限；选取不同剂量（100%、50%、25%、10%）丝素膜浸提液对家兔进行溶血试验，溶血率为4.02%、2.57%、1.56%、1.01%，均小于5%。综上所述，本研究制备的再生丝素膜状材料，具有良好的生物安全性。
Abstract
Water-fast silk fibroin films were created from regenerated silk fibroin by drying at a controlled temperature and humidity and degenerating with ethanol. To evaluate the biosafety of these silk fibroin membranes, cytotoxicity test in vitro, general allergic, delayed- type hypersensitivity assay, acute toxicity, pyrogenic reaction and hemolytic test on erythrocytes of rabbits were conducted. The results showed that: after in vitro cultured fibroblasts were treated with silk fibroin membrane leaching liquor, they proliferated appropriately at 92.3%, indicating the cytotoxicity of silk fibroin membrane leaching liquor was grade I; after repeated injections of 100% silk fibroin membrane leaching liquor, no mice presented generalized anaphylaxis; all the mice that received injections of 100% silk fibroin membrane leaching liquor survived, no acute general toxic reactions; the body weight of mice gradually increased within 7 days. After injection of 100% silk fibroin membrane leaching liquor into the ear vein of Japanese big eared rabbits, the body temperature of per rabbit increased by 0.2-0.5℃, but did not exceed the upper limit of 0.6, and 1.4℃ in total. The hemolysis test showed that the hemolytic rates of 100%, 50%, 25%, and 10% silk fibroin membrane leaching liquor were 4.02%, 2.57% , 1.56% , and 1.01% ，respectively, all rates were lower than 5% . In short, the data indicate that regenerated silk fibroin films exhibit good biosafety.
关键词
再生丝素;蛋白;膜状材料;生物安全性;
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Key words
regenerated silk fibroin;silk fibroin;membrane;biosafety;
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参考文献
[1]朱希山, 曾丽芬, 赵春华.组织工程化人工皮肤研究的新进展[J].中国组织工程研究与临床复, 2007, 11(6):1145-1148
[2]汝玲, 黄毅萍, 陈萍, 等.蚕丝素蛋白最新研究进展[J].化学推进剂与高分子材料, 2007, 5(4):26-36
[3]赵广建, 赵耀.人工皮肤替代物的材料种类及其特征[J].中国组织工程研究与临床复, 2010, 14(29):5467-5470
[4]裴莹, 郑学晶, 汤克勇.胶原及明胶基天然高分子复合材料研究进展[J].高分子通报, 2010, 2(2):58-68
[5]谢长琼, 周元林, 马佳俊.聚己内脂改性研究进展[J].塑料科技, 2009, 37(4):100-104
[6]许海叶, 徐卫林, 刘秀英.非水溶性丝素粉体与聚氨酯再生丝素膜的性能[J].纺织学报, 2008, 29(11):16-21
[7]张加忠, 陈建勇, 冯新星, 等.聚乳酸改性丝素膜的性能研究[J].功能材料, 2007, 12(38):2048-2051
[8]吴瑢蓉, 王敏超, 曹厚宝, 等.交联剂对再生是丝素蛋白结构域性能的影响[J].高科技纤维与应用, 2012, 37(06):66-69
[9]Altman GH, Diaz F, Jakuba C, et al.Silk-based biomaterials[J].Biomaterials, 2003, 24(3):401-416
[10]Minoura N, Tsukada M, Nagura M.Physico-chemical properities of silk fibroin membrane as biomaterial[J].Biomaterials, 1990, 11(6):430-434
[11]黄训亭, 邵正中, 陈新.天然蚕丝与丝素蛋白多孔膜的生物降解性研究[J].化学学报, 2007, 65(22):2592-2596
[12]周婵, 刘彬, 王介平, 等.家蚕丝素蛋白在组织工程中的应用研究进展[J].中国畜牧兽医杂志, 2013, 23(49):91-94
[13] 代时春, 徐水, 李慧, 等.家蚕后部丝腺丝素蛋白的制备[J].中国组织工程研究与临床康复, 2009, 13(12):2269-2272
[14] Vepari C, Kaplan DL.Silk as a biomaterial[J].Prog Polym Sci, 2007, 12(32):991-1007
[15]陈春花, 朱良均, 邹凤竹.丝素壳聚糖共混膜性能测定及制备配方探讨[J].蚕业科学, 2007, 33(1):79-84
[16] 曹惠.自透析法再生丝素纤维的制备及其结构性能研究[D].硕士学位论文. 苏州：苏州大学, 2005.
[17]周婵, 徐水, 成国涛, 等.丝素纳米凝胶共混膜的生物安全性评估试验[J].蚕业科学, 2011, 3(37):466-472
[18]张瑞, 韩宝三, 彭承宏.壳聚糖球形多孔微载体的组织相容性评价[J].内蒙古医学院学报, 2011, 2(33):119-124
[19]关静, 高萍, 黄姝杰, 等.聚乙烯醇水凝胶的生物相容性评价[J].中国组织工程研究与临床康复, 2009, 12(13):2227-2231
[20]黄泉, 杨吉成, 缪竟诚.丝素蛋白材料生物相容性评价研究进展[J].国际生物医学工程杂志, 2007, 30(3):163-166
[21] 侯春春.丝素蛋白/碳酸钙复合支架材料的制备及其理化性能、生物相容性研究[D].硕士学位论文. 重庆：西南大学, 2011.
[22]Minoura N, Aiba S, Higuchi M, et a.Attachment and growth of fibroblast cells on silk fibroin[J].Biochem Biophys Res Commun, 1995, 208(2):511-516
[23]张道明, 涂献玉, 余杨, 等.炭纤维增强壳聚糖内固定棒的家兔热原试验[J].长江大学学报 自科版, 2005, 12(2):339-340
[24] 国家食品药品监督管理局．化学药物刺激性、过敏性和溶血性研究技术指导原则[S].2005，5-27.
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