羟基磷灰石/左旋聚乳酸对人骨髓有核细胞毒性的实验  

Hydroxyapatite/poly-L-lactide for cytotoxicity of human bone marrow karyote

作　　者：于博[1] 靳安民[1] 吴广森[1] 汤善华[1] 梁伦高[1] 田京[1] 张辉[1] 柳晓秋[2] 胡国栋[3] 

机构地区：[1]南方医科大学珠江医院脊柱骨病科,广东省广州市510282 [2]南方医科大学珠江医院神经生物研究所,广东省广州市510282 [3]南方医科大学南方医院呼吸科,广东省广州市516516

出　　处：《中国临床康复》2005年第46期42-43,T0003,共3页Chinese Journal of Clinical Rehabilitation

基　　金：广东省自然科学基金重点攻关课题(2001A302020205)~~

摘　　要：目的:评价不同浓度羟基磷灰石/左旋聚乳酸与人体骨髓有核细胞复合体体外培养时的细胞毒性及生物相容性。方法:实验于2003-03/05在南方医科大学珠江医院神经外科实验室完成。将自行制备的羟基磷灰石/左旋聚乳酸配制成不同浓度(4,2,1,0.5,0.1g/L)与来自临床志愿者骨髓的人体骨髓有核细胞进行复合体外培养,对照组单纯接种细胞。采用相差显微镜逐日观察细胞生长及与生物材料的附着情况。并采用四甲基偶氮唑盐微量酶反应比色法于不同培养时间(2,4,6,8d)测定细胞增值率(以酶联仪上570nm处测吸光度值表示),以反映羟基磷灰石/左旋聚乳酸复合材料对骨髓有核细胞的毒性作用。结果:①对照组及不同浓度(4,2,1,0.5,0.1g/L)羟基磷灰石/左旋聚乳酸细胞培养溶液在培养2,4,6,8d时所测得光密度值均比较接近,差异无显著性(F=0.070～0.255,P=0.929～0.996)。②相差显微镜观察细胞增殖形态学变化见骨髓基质细胞与羟基磷灰石/左旋聚乳酸材料复合培养组早期可见培养板内细胞增多,少量细胞向材料移行贴附于材料周边,部分细胞直接贴附于材料表面,随复合培养时间延长,附着的细胞增多,无细胞衰老及异常分裂现象。结论:从形态学观察羟基磷灰石/左旋聚乳酸对人体骨髓有核细胞增殖无明显影响,未见明显细胞毒性。AIM： To evaluate the cytotoxicity and biocompatibility of hydroxyapatite/ poly-L-lactide （HA/PLEA） composite of different concentration when it is in vitro cultured with human bone marrow karyote. METHODS： The study was made in the Neurosurgery Laboratory of Zhujiang Hospital of Southern Medical University between March and May 2003. Human Bone marrow stramal cells were cultured in vitro with HA/ PLLA composite of different concentration of （4,2,1,0.5,0.1 g/L）, the cells were inoculated only in the control group. The cell growth and adhesion of biomaterial were observed with phase contrast microscope day by day. The rate of cell proliferation was detected with met, lyl-thiazol-tetrazolium （MTT） colorimetry at different time points （2, 4, 6 and 8 days）, which were expressed by the absorbance at 570 nm on enzyme-link meter, to reflect the cytotoxicity of HA/PLIA composite to bone marrow karyote. RESULTS： ① The absorbances of HA/PLLA cell culture medium of different concentration （4, 2, 1, 0.5, 0.1 g/L） and control group obtained at 2, 4, 6 and 8 days were close, and there were insignificant differences （F =0.070-0.255, P=0.929-0.996）. ② The morphological changes of cell proliferation observed under phase contrast microscope showed that in the bone marrow stromal cell and HA/PLIA composite culture group, there was increase of cells in culture plate, a few cells moved towards the material and adhered to the marginal region, and part of the cells adhered to the surface of the material directly; With the prolongation of co-cultured time, the adherent cells were increased, no cell aging and abnormal splitting were observed. CONCLUSION： It is morphologically observed that HA/PLLA has no obvious effect on the proliferation of human bone marrow karyote, and there is no obvious cytotoxicity.

关 键 词：羟基磷灰石类 骨髓细胞 生物相容性材料 

分 类 号：R318.08[医药卫生—生物医学工程]