再生丝素膜的生物安全性评估试验  

Evaluation of the Biological Safety of SF Membrane

作　　者：周婵[1] 王介平[1] 吕金凤[1] 王海燕[1] 曾秀[1] 

机构地区：[1]重庆市畜牧科学院,重庆402460

出　　处：《中国农学通报》2014年第33期156-161,共6页Chinese Agricultural Science Bulletin

基　　金：国家自然科学基金资助项目"phrinB2/蚕丝组织工程神经的研究"(31271036);重庆市基本科研业务费资助项目"桑蚕中肠细胞原代培养研究"(2012csfc-jbky-00909)

摘　　要：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,7天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,研究制备的再生丝素膜状材料,具有良好的生物安全性。Water-fast silk fibroin films were created from regenerated silk fibroin by drying at a controlledtemperature and humidity and degenerating with ethanol. To evaluate the biosafety of these silk fibroinmembranes, 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 proliferatedappropriately at 92.3%, indicating the cytotoxicity of silk fibroin membrane leaching liquor was grade I; afterrepeated 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 generaltoxic reactions; the body weight of mice gradually increased within 7 days. After injection of 100% silk fibroinmembrane leaching liquor into the ear vein of Japanese big eared rabbits, the body temperature of per rabbitincreased by 0.2-0.5℃, but did not exceed the upper limit of 0.6, and 1.4℃ in total. The hemolysis test showedthat 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 thatregenerated silk fibroin films exhibit good biosafety.

关 键 词：再生丝素 蛋白 膜状材料 生物安全性 

分 类 号：S881.3[农业科学—特种经济动物饲养] R318.08[农业科学—畜牧兽医]