纳米羟基磷灰石/聚氨基酸复合材料体外成骨样细胞相容性评价

Biocompatibility Evaluation of Nano-hydroxyapatite/Poly-amino Acid(n-HA/PAA) Composite with Osteoblast-like Cells in Vitro

为评价自制纳米羟基磷灰石/聚氨基酸(n-HA/PAA)复合材料体外细胞相容性,采用MG63成骨样细胞与材料共同培养,以n-HA为对照进行以下实验:①细胞形态学观察;②细胞吸光度值的MTT检测;③细胞碱性磷酸酶(ALP)活力检测;④细胞骨钙素(OCN)含量的酶联免疫法(ELISA)检测;⑤流式细胞仪检测细胞周期及细胞凋亡情况。结果表明,n-HA/PAA复合材料对MG63细胞形态无明显影响,材料周围及表面细胞贴壁生长良好,并随时间的延长细胞逐渐增多。MTT检测细胞吸光度值随培养时间的延长而逐渐增加,且细胞增殖明显,实验组与对照组间各时间点比较无明显差异(P>0.05)。细胞ALP活力及OCN含量随培养时间的延长逐渐增加,实验组与对照组比较无明显差异(P>0.05)。两组材料对细胞的生长周期及凋亡均无明显影响,且两组细胞周期及凋亡细胞各自比较无明显差异(P>0.05)。

To evaluate the biocompatihility of self-made nancrhydroxyapatite/poly-amino acid（n-HA/PAA） composite in vitro, the MG63 cell line was used to be cultured with n-HA and n-HA/PAA composite, n-HA was used as control group to carry out the following experiments： ① The cells morphologic observation;② The detection of the OD value by MTT assay; ③ The ALP activity test; 0 The OCN level test; ④ The detection of cell cycle and apoptosis by flow cytometry. The results show that there are no obvious cell morphologic changes in the n-HA and n-HA/ PAA groups, and the adherent MG63 cells around n-HA and n-HA/PAA and on their surface grow well. MTT assay reveals that the MG63 cells had obvious proliferation, the OD value increases gradually with the increasing time of culture in the two groups, and there is no significant difference（P）0. 05） in the two groups at all time points. The ALP activity and OCN level of MG63 cells increase gradually with the increasing time of culture in the two groups, and there is no significant difference（P〉0. 05） in the two groups at all time points. The n-HA and n-HA/PAA composite can＇t inhibit the cell cycle of MG63 cells and promote the apoptosis, and there is no obvious difference（P〉0. 05） of the cell cycle and apoptosis in the two groups.