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目的研究不同蚕丝蛋白材料对鼠胚表皮细胞的毒性及其影响细胞增殖的因素。方法采用组织块培养法体外原代培养鼠胚表皮细胞,用直接接触法和浸提液法体外培养鼠胚表皮细胞,MTT比色法检测其在不同蚕丝蛋白材料上的增殖活力和细胞相对增殖率,进行毒性分析;并用活细胞计数法观察不同蚕丝蛋白材料对鼠胚表皮细胞生长的影响。结果鼠胚表皮细胞在1、2、4、5、7、8、10号各型蚕丝蛋白材料上生长,其增殖活力与细胞对照组相当(P>0.05);在3、6、9号化学交联或HY-823交联型蚕丝蛋白材料上生长,其增殖活力均低于细胞对照组(P<0.05);而在11号蚕丝蛋白材料其增殖活力显著低于细胞对照组(P<0.01)。1、7号丝胶材料细胞毒性分级为0级(无毒),2、3、4、5、6、8、9、10号各型蚕丝蛋白材料细胞毒性分级为1级(轻微毒),而11号蚕丝蛋白材料细胞毒性分级为2级(中度毒)。活细胞计数结果与MTT检测结果基本相一致。结论除11号蚕丝蛋白材料外,1~10号各型蚕丝蛋白材料无明显细胞毒性,能支持鼠胚表皮细胞正常生长,具有良好的细胞相容性。
Objective To study the toxicity of different silk fibroin materials on mouse embryonic epidermal cells and the factors affecting cell proliferation. Methods The primary cultured human embryonic epidermal cells were cultured with tissue culture method. The direct contact method and the extraction method were used to culture the epidermal cells in vitro. MTT assay was used to detect the proliferative activity and relative proliferation of fibroblasts in different silk fibroin materials The toxicity was analyzed. The effects of different silk fibroin materials on the growth of mouse embryonic epidermal cells were observed by viable cell counting. Results The mouse embryonic epidermal cells grew on silk fibroin materials 1, 2, 4, 5, 7, 8 and 10 with a similar proliferative activity as that of the cell control group (P> 0.05) (P <0.05). However, the proliferation activity of silk fibroin No. 11 was significantly lower than that of the cell control group (P <0.01), and the proliferation activity of HY-823 cross-linked silk protein material was lower than that of the cell control group ). The cytotoxicity grade of No. 1 and No. 7 sericin materials was grade 0 (non-toxic). The cytotoxicity grade of various types of fibroin materials of No. 2,3,4,5,6,8,9,10 were classified as Grade 1 (mild toxicity) The No. 11 silk protein material cytotoxicity grade 2 (moderate poison). Live cell count results and MTT test results are basically the same. Conclusion Except No. 11 silk protein material, all types of silk fibroin materials No. 1-10 have no obvious cytotoxicity and can support the normal growth of mouse embryonic epidermal cells with good cell compatibility.