三维多孔β-磷酸三钙支架与骨髓基质细胞构建组织工程骨的生物相容性及显微形态学观察  

Biological compatibility and micromorphologic observation of three dimensional porous beta-tricalcium phosphate scaffolds and mesenchymal stem cells in constructing tissue engineered bone

作　　者：程晓兵[1] 马秦[1] 侯锐[1] 杨耀武[1] 毛天球[1] 敖建华[1] 

机构地区：[1]解放军第四军医大学口腔医学院颌面外科,陕西省西安市710032

出　　处：《中国组织工程研究与临床康复》2007年第9期1640-1643,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

摘　　要：目的:定量及显微观察三维多孔β-磷酸三钙支架在构建组织工程骨时的组织相容性。方法:实验于2002-07/2005-07在第四军医大学口腔医学院颌面外科组织工程实验室完成。将兔的骨髓基质细胞,按一定量分置在培养板的各孔内,将多孔β-磷酸三钙支架材料置入孔内共培养,采用MTT法和碱性磷酸酶试剂盒检测骨髓基质细胞活性,共聚焦显微镜观察其在β-磷酸三钙支架表面的微观形态。结果:①倒置显微镜下观察接种前的细胞形态基本均匀一致。②接种后6d扫描电镜观察显示,材料表面可见较多细胞和细胞合成的丝状细胞外基质。③细胞接种后6d激光共聚焦显微镜观察显示,细胞数量明显增多,基本长满材料的表面,同时在孔隙中,也有大量细胞长入。④β-磷酸三钙对骨髓基质细胞的增殖作用:培养第5天,β-磷酸三钙组对骨髓基质细胞的增殖作用(A)低于对照组,差异有显著性意义(分别为0.47±0.02,0.52±0.02,t=2.281,P<0.05)。⑤β-磷酸三钙对骨髓基质细胞的碱性磷酸酶的活性比较:在骨髓基质细胞诱导培养5,7,9d,β-磷酸三钙组碱性磷酸酶的活性(A)明显低于对照组,差异有显著性意义(分别为5.18±0.31,6.05±0.54;12.35±0.62,14.17±0.70;15.68±0.87,17.13±1.14,t=3.302,6.025,6.019,P<0.05)。结论:三维多孔β-磷酸三钙支架有良好的组织相容性,为其应用于临床提供了可靠的实验依据。AIM： A quantitative and micromorphologic observation of tissue compatibility of three-dimensional porous beta-tricalcium phosphate （β-TCP） scaffold in tissue engineered bone. METHODS： From July 2002 to July 2005 the experiment was performed in Tissue Engineering Laboratory, Department of Oral and Maxillofacial Surgery, Stomatological College, Fourth Military Medical University of Chinese PLA. Mesenchymal stem cells （MSCs） of rabbit was placed in the wells of microplate quantitatively, then β-TCP scaffolds were put into the wells for coculture. Activity of MSCs was evaluated by MTT assay and alkaline phosphatase （ALP） kit, and the micromorphology of MSCs was observed on the surface of β-TCP by confocal microscope. RESULTS： （1）The osteoblasts had uniformity appearance under the inverted microscope before inoculation. （2） days after inoculation there were many osteoblast and filarious extracellular matrix under the scanning electron microscope. （3）6 days after inoculation, there were large number of osteoblast on the surface and in the pore of the scaffold material under the laser confocal microscopy. （4）Proliferative effect of β-TCP on MSCs： On the 5^th day, the proliferative effect （A） of β-TCP on MSCs in the β-TCP group was lower than that in the control group. There were significant differences （0.47±0.02,0.52±0.02, t=2.281, P 〈 0.05）. （5）Activity of alkaline phosphatase in osteoblasts： The activity of alkaline phosphatase （A） in β-TCP group was obviously lower than that in the control group at days 5, 7 and 9. There were significant differences （5.18±0.31, 6.05±0.54; 12.35±0.62,14.17±0.70; 15.68±0.87,17.13±1.14, t =3.302,6.025,6.019, P 〈 0.05）. CONCLUSION： Three dimensional porous β-TCP scaffold has favourable tissue compatibility, and these experiments offer a credible experimental evidence to clinical application of β-TCP scaffold.

关 键 词：组织工程 磷酸三钙 骨髓基质细胞 支架 

分 类 号：R318.08[医药卫生—生物医学工程]