重组类人胶原蛋白Ⅱ膜的生物相容性

Biocompatibility of recombinant human-like collagen Ⅱ membranes crosslinked with glutaraldehyde

目的探讨用戊二醛交联得到的重组类人胶原蛋白Ⅱ（recombinant human-like collagenⅡ,RHLCⅡ）膜的生物相容性。方法用浓度为0.005 mol/L和0.01 mol/L的戊二醛分别交联RHLCⅡ得到两组RHLCⅡ膜（0.005 mol/L组和0.01 mol/L组）;把成纤维细胞接种到两组RHLCⅡ膜上,用倒置显微镜和扫描电子显微镜观察细胞贴附情况;用MTT法分析成纤维细胞在两组RHLCⅡ膜上的生长和增殖情况;通过细胞毒性实验评价两组RHLCⅡ膜的细胞毒性。结果当成纤维细胞培养到6 h时,均能贴附于两组RHLCⅡ膜上;培养到96 h时,两组RHLCⅡ膜上的成纤维细胞均融合成片并形成多层细胞;经MTT法分析,细胞接种后96 h,成纤维细胞在两组RHLCⅡ膜上均能实现增殖;0.01 mol/L组比0.005 mol/L组细胞毒性要大,但两组在各时间段,毒性评定均在0-1级,细胞毒性实验合格。结论戊二醛浓度为0.005 mol/L和0.01 mol/L的RHLCⅡ膜具有良好的生物相容性。

Abstract： Aim To study the biocompatibility of recombinant human-like collagen Ⅱ （ RHLC Ⅱ ） membranes crosslinked with glutaraldehyde. Methods RHLC Ⅱwas crosslinked with different concentrations （0. 005 mol/L and 0. 01 mol/L） of glutaraldehyde to form two groups of RHLC Ⅱ membranes. Fibroblasts were cultured as seeding cells on RHLC Ⅱ membranes, then the cells-material complex was observed with contrast and scanning electronic microscope. The proliferation of fibroblasts were tested by MTT. RHLC Ⅱ membranes＇ cytotoxicity was evaluated. Results After 6 h cultivation, fibroblasts adhered to both groups. Fibroblasts formed several confluent layers on two groups of RHLC membranes in 96 h. Fibroblasts can proliferate on two kinds of RHLC Ⅱ membranes within 96 h. Compared with RHLC Ⅱ membranes crosslinked by 0. 005 mol/L glutaraldehyde, RHLC Ⅱ membranes with 0. 01 mol/L glutaraldehyde have stronger cytotoxicity. But reaction scales of both RHLC Ⅱ membranes 0 - 1 level. Cytotoxicity of RHLC Ⅱ membranes met the requirement of standards. Conclusion RHLC Ⅱ membranes crosslinked with 0. 005 mol/L and 0.01 mol/L glutaraldehyde have good biocompatibility and can become ideal materials for skin tissue engineering.