蛋白石/聚左旋乳酸复合材料与人成骨细胞的相容性  

Compatibility of opal shale/poly-L-lactic acid composite with human osteoblasts

作　　者：管映亭[1] 陶用珍[2] 金志浩[1] 

机构地区：[1]西安交通大学材料科学与工程学院金属材料强度国家重点实验室 [2]西安交通大学生命科学与技术学院生物科学与工程系,陕西省西安市710049

出　　处：《中国临床康复》2006年第45期68-70,共3页Chinese Journal of Clinical Rehabilitation

摘　　要：目的:在体外细胞培养条件下,从细胞形态和细胞增殖方面观察和评价蛋白石/聚左旋乳酸复合材料与人成骨细胞的生物相容性。方法:实验于2004-09/2005-11在香港理工大学应用生物和化学系细胞实验室及西安交通大学生命科学与技术学院生物科学与工程系完成。参照美国药典对医用高分子材料制作浸提液的方法,以无菌生理盐水作为浸提介质,对蛋白石/聚左旋乳酸复合材料和聚左旋乳酸材料分别制取浸提液。测定提取液及其稀释液(1∶1,1∶4,1∶16,1∶32,1∶64,1∶128,1∶256)的pH值,并在加有此浸提液的培养基中培养成骨细胞,空白对照组加等量无菌生理盐水,在倒置相差显微镜下观察细胞的生长情况;选用四甲基偶氮唑盐(MTT)法测定细胞的增殖情况。结果:①蛋白石/聚左旋乳酸复合材料提取液及其稀释液的pH值高于相应浓度聚左旋乳酸材料,说明复合材料释酸量减少,有利于避免因聚左旋乳酸释酸而引起的各种不良反应。②倒置显微镜观察显示,加入蛋白石/聚左旋乳酸复合材料浸提液的培养皿中成骨细胞的增殖与空白对照组差异无显著性意义,均未发现细胞衰老及异常分裂现象。③MTT法测定表明不同浓度的蛋白石/聚左旋乳酸复合材料和聚左旋乳酸材料浸提液加入后,细胞的增殖不论是与空白对照比较,还是它们二者之间相互比较,差异均无显著性意义(P>0.05)。以美国药典中细胞相对增殖率与细胞毒性分级的关系,蛋白石/聚左旋乳酸复合材料为USP毒性0级,即无毒级。结论:蛋白石/聚左旋乳酸复合材料与聚左旋乳酸材料表现出相似的细胞相容性。考虑到蛋白石的加入使聚左旋乳酸的释酸量减少,再加上蛋白石的负离子释放功能,蛋白石/聚左旋乳酸复合材料必将在组织工程领域存在着广阔的应用前景。AIM： This paper reports and evaluates the biocompatibility of the opal shale/poly-L-lactic acid （PLLA） composite with human osteoblasts by observing the morphology and proliferation of in vitro cultured cells. METHODS： The experiment was carried out in the Cell Laboratory of Applied Biology and Chemical Technology Department, Hong Kong Polytechnic University and Department of Biological Science and Engineering, School of Life Science and Technology, Xi＇an Jiaotong University from September 2004 to November 2005. Referring to the method of solution extraction from medical macromolecule materials by the US Pharmacopoeia （USP）, opal shale/PLLA composite and PLLA were respectively extracted by using asepsis saline as medium. And pH values of the extracted solution and their diluted ones （1：1, 1：4, 1：16, 1：32, 1：64, 1：128, and 1：256） were tested. Then osteoblasts were added into the cultivation medium, while equal volume of saline was injected in control group. Cellular morphology was observed by inverted phase contrast microscope. The proliferation of osteoblasts was quantitively evaluated by 3-[4, 5-dimethyl thiazol-2-yl]-2, 5-diphenyl tetrazolium bromide （MTT） colorimetry method. RESULTS： ①The pH values of extracts and their diluted ones from composite were higher than those obtained from PLLA. This meant that the release of acid from composite was reduced and harmful effects from the generation of acid was avoided. ②Through observation via inverted phase contrast microscope, the proliferation of osteoblasts growing in the medium with composite were almost the same as those growing without composite. No cell aging or abnormal cell division was observed. ③MTT study showed that extracts from opal shale/PLLA composite or PLLA had no statistical difference for the growth of osteoblasts and neither of them had statistical difference from contrast （without extracts） （P 〉 0.05）. The cell toxicity of opal shale/PLLA composite was graded 0 according to cytotoxicity grade

关 键 词：材料试验 聚合物/代谢 乳酸/代谢 成骨细胞 生物相容性材料 

分 类 号：R318[医药卫生—生物医学工程]