脱细胞软骨生物支架材料的生物学特性

目的 探讨脱细胞软骨生物支架材料（ACM）在细胞毒性、溶血试验、急性全身毒性等方面的生物学特性：方法①ACM的制备：猪膝关节软骨冻干加工为粉末，胰酶消化，曲拉通洗脱，蒸馏水洗净冻干，紫外线照射（UVI）后成型；③细胞毒性测定：材料浸提液培养细胞进行细胞形态人体观察，MTT法观察细胞活性；③急性全身毒性反应：材料浸提液注射入SD大鼠腹腔观察材料对动物表现及体重变化的影响；④溶血试验：材料浸提液与稀释动物鲜血混合观察红细胞溶解情况，492nm下检测OD值计算相对溶血率；结果 ①细胞毒性试验：24h、48h、72h各时间段内三组细胞OD值两两比较（P〉0．05），无显著差异，ACM细胞毒性为0级；②动物急性毒性实验：Ⅰ生物材料处理组和Ⅱ生理盐水处理组对动物体重影响没有差异（P〉0．05），Ⅰ生物材料处珊组和Ⅲ苯酚处理组对动物体重有显著差异（P〈0．05）；③溶血试验：材料相对溶血率为2.92％，低于5％的标准，尢明显溶血现象；结论ACM存细胞毒性、溶血实验、动物急性毒件反应方面符合软骨组织工程中对于支架材料的要求，提示支架材料何良好的牛物相容性和安全性。

Biological properties researches of acellular cartilage material

Objective To investigate the biocompatibility of the acellular cartilage material in test for in vitro cytotoxicity, acute systemic toxicity and test of hemolysis; Methods ①ACM preparation： The articular cartilage of pigs was processed into powder. The components of cells were eliminated by Trypsin. TritonX-100 and distilled water. Finally the powder was freezed out by freezedryer and irradiated by ultraviolet;②Test for in vitro cytotoxicity： The BMSCs were cultured in the special L-DMEM that ACM were dipped in and then observed through microscope and MTT; ③Test for acute systemic toxicity： The changes of the behavior and the avoirdupois of the SD rats were evaluated after the special L- DMEM was injected into the SD rats by intraperitoneal injection; ④Test of hemolysis： The grade of hemolysis was made in the macroscopical phenomenon and the measures of optical density at 492nm; Results ①Test for in vitro cytotoxicity： At every time interval, the OD scores of three groups were not significant differences （P〉0.05）.The grade of the cytotoxicity of the ACM was 0;②Test for acute systemic toxicity： The avoirdupois scores of both group Ⅰ and Ⅱ were not statistically significant differences （P〉0.05）, but the differences between group Ⅰ and Ⅲ were statistically significant （P〈0.05）;③Test of hemolysis： Hemolysis Percentage was 2.92%; Conclusion ACM accords with the biomaterial standards of cartilage tissue engineering in test for in vitro cytotoxicity, test of hemolysis and test for acute systemic toxicity.