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中国农学通报 ›› 2014, Vol. 30 ›› Issue (33): 156-161.doi: 10.11924/j.issn.1000-6850.2014-2155

• 生物技术科学 • 上一篇    下一篇

再生丝素膜的生物安全性评估试验

周婵,王介平,吕金凤,王海燕,曾秀   

  1. 重庆市畜牧科学院,重庆市畜牧科学院,重庆市畜牧科学院,重庆市畜牧科学院,重庆市畜牧科学院
  • 收稿日期:2014-08-07 修回日期:2014-08-07 接受日期:2014-09-05 出版日期:2015-01-08 发布日期:2015-01-08
  • 通讯作者: 曾秀
  • 基金资助:
    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)。

Evaluation of the Biological Safety of SF Membrane

  • Received:2014-08-07 Revised:2014-08-07 Accepted:2014-09-05 Online:2015-01-08 Published:2015-01-08

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,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。

关键词: 多聚体, 多聚体

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.