纳米羟基磷灰石/聚酰胺66与人胚胎成纤维细胞的生物相容性研究

目的:研究新型骨修复重建复合材料纳米羟基磷灰石/聚酰胺66(n-HA/PA66)对人胚胎成纤维细胞可能存在的细胞毒性,为该材料应用于临床提供实验依据.方法:采用人胚胎成纤维细胞,经材料及材料浸提液分别与细胞共培养等方式对n-HA/PA66进行细胞毒性测试,细胞形态观察法观察各组细胞在24h、48h、72h、5天各时相点的细胞形态学变化,细胞生长抑制法(MTT比色法),测定各组1,2,4,7天细胞的相对增殖率.结果:材料表面的细胞黏附生长良好,各浸提液组和直接接触组分别与阴性对照组进行两两比较,各组对细胞的增殖均无显著影响(P＞0.05),各浸提液组和直接接触组分别与阳性对照组进行两两比较显示有显著差异(P＜0.05),材料的细胞毒性为0～1级.结论:体外试验结果显示,n-HA/PA66与人胚胎成纤维细胞相容性好,符合GB/T16886.5-1997-ISO 10993-5:1992规定的医用材料相应标准1].

Study on biocompatability of nano-hydroxyapatite crystals and polyamide 66 composite with human embryonic fibroblast

Objective:To evaluate the biocompatability of nano-hydroxyapatite crystals and polyamide composite(n-HA/PA66) with the human embryonic fibroblast(HEFs),to provide an experimental evidence for its clinical application.Methods:The morphological observation was made and pictures of HEFs were taken 1d,2d,4d,and 7d after co-culture with extract of n-HA/PA66,and direct contact with n-HA/PA66.To determine absorbance of every hole under 500 nm with enzyme linked immunity instrument after 1d,2d,4d,and 7d of contact of n-HA/PA66 extract with HEF cells,and direct contact with n-HA/PA66.In the meanwhile calculate the relative multiplication rate of cells,and evaluate them by six degree tests for cytotoxicity.Results:The HEFs in direct contact culture with composite were attached and proliferated well on the materials surface.The growth and proliferation of cultured cells was not significantly inhibited between every composite group with the negative group ( P >0.05).There was significant difference between every composite group with the positive group( P <0.05).In this experiment the toxicity of n-HA/PA66 to HEF cells ranges from zero to one degree.Conclusion:n-HA/PA66 have good biocompatibility with human embryonic fibroblast(HEFs) in vitro,up to the standard of International Organization for Standardization(ISO).