二氧化锆梯度复合羟基磷灰石体外激活末梢血单个核细胞的实验研究

Peripheral blood mononuclear cells activated by graded zirconia-hydroxyapatite composite in vitro

目的:体外检测二氧化锆(ZrO2)梯度复合羟基磷灰石(HAP)材料浸提液对健康青年人外周血单个核细胞(PBMC)的激活,来评价其免疫原性。方法:以二氧化锆(ZrO2)单纯复合羟基磷灰石(HAP)为对照材料,制备两种材料浸提液并分离健康年轻人PBMC。MTT法检测两种材料浸提液刺激PBMC转化的作用;ELISA法检测两种材料浸提液刺激PBMC分泌TNFα和IL-6的作用;流式细胞术检测两种材料浸提液刺激PBMCCD69、CD25的作用。结果:培养5d,单纯复合组对PBMC的增殖作用弱于对照组(P<0.05),而梯度复合组与阴性对照组比较无差异;LPS共培养24h,两材料组TNFα和IL-6水平差异显著(P<0.05);PHA作用24h,两材料组PBMC中CD69和CD25阳性细胞率差异显著(P<0.01)。结论:ZrO梯度复合HAP生物陶瓷材料免疫原性弱,对PBMC的激活程度较轻。

AIM： To evaluate the immunogenicity of a novel orthopedics materials （graded zirconia - hydroxyapatite composite） in vitro by using peripheral blood mononuclear cells （PBMCs） from healthy young people, and simple zirconia- hydroxyapatite composited material was used as control materials. METHODS： Proliferation of PBMCs cultured in different liquid after 5 days was measured by MTr methods. ELISA was used to detect TNF -α and IL - 6 concentration in the supernatant of PBMCs cultured in the extracts after 24 hours. Flow cytometery was used to measure CD69 and CD25 in activated PBMCs cultured in the extracts of the two kinds of materials after 24 hours. RESULTS ： The proliferation rate of the simple composite group was significantly lower than that in negative group （ P 〈 0.05）, but there was no difference between the graded composited group and negative group. After 24 hours culture with LPS, the concentrations of TNF - α and IL - 6 in the simple composited group were significantly higher than that in graded composited group, respectively （ P 〈 0.05）. Cultured with PHA for 24 hours, the ratio of CD69 and C1Y25 positive PBMCs in the simple composited group was all significantly higher than that in graded composited material group （P 〈 0.01）. CONCLUSION ： The number of PBMCs activated by the graded composited material is less than the simple composited material and the immunogenicity of the graded composited material is lower than the simple composited material.