新型β-磷酸三钙的制备与成骨能力的实验研究  

An experimental study on the preparation of a novelβ-tricalcium phosphate and its osteogenic potential as scaffold for bone tissue engineering

作　　者：刘广鹏[1] 赵莉[1] 刘波[1] 袁捷[1] 崔磊[1] 刘伟[1] 曹谊林[1] 

机构地区：[1]上海组织工程研究与开发中心,200235

出　　处：《中华创伤骨科杂志》2006年第8期755-759,共5页Chinese Journal of Orthopaedic Trauma

基　　金：国家"973"组织工程基本科学问题项目(G1999054304);国家"863"组织工程化骨构建技术与产品开发项目(2002AA20511)

摘　　要：目的采用有机泡沫浸渍法制备多孔β-磷酸三钙(β-TCP)生物陶瓷,探讨其作为骨组织工程支架材料的可能性。方法有机泡沫浸渍法制备β-TCP支架材料,接种人骨髓基质干细胞,体外成骨诱导培养。相同条件下培养的单层细胞作为对照组。采用扫描电镜和噻唑蓝比色法观察细胞在材料上的增殖能力,同时测定碱性磷酸酶活性和骨钙蛋白含量来观察细胞在材料上的成骨分化能力。将体外培养7 d的细胞材料复合物和单纯材料回植裸鼠皮下,分别于术后4、8、12周取材观察异位成骨情况。结果扫描电镜观察显示实验组细胞在材料上增殖良好,MTT结果与对照组无明尼差异,但碱性磷酸酶活性和骨钙蛋白含量均高于单层培养的细胞。细胞材料复合物在裸鼠皮下4周已经成骨,并随时间延长骨量增多。单纯材料组在各时间点均未成骨。结论有机泡沫浸渍法制备的多孔β-TCP生物陶瓷具有良好的支持成骨能力,是一种较理想的骨组织工程支架材料。Objective To prepare porous β-tricaleium phosphate（β-TCP） ceramics by the polymeric sponge method and investigate its potential as scaffold for bone tissue engineering. Methods Porous β-TCP ceramics were prepared by the polymeric sponge method. Human bone marrow stromal cells （BMSCs） at passage two were seeded onto the ceramics and cultured in osteogenic media in vitro, with monolayer cells treated under the same condition as control. Cell proliferation and osteogenic differentiation on the scaffold were evaluated by means of scanning electron microscope （SEM） observation, tetrazolium salt （MTT） assay, and measurement of alkaline phosphatase （ALP） activity and osteocalcin （OCN） content. Moreover, after seven days＇ culture, the scaffolds with and without human BMSCs were implanted subcutaneously into nude mice. Samples were harvested at four, eight, and 12 weeks after treatment respectively to observe the heterotopic ossification. Results SEM observation showed that human BMSCs attached well on the scaffolds and proliferated rapidly. Compared with the cells in monolayer culture, they mainta/ned higher ALP activity and OCN content. On the scaffolds with human BMSCs, new bone formed after four weeks of implantation, and osteogenesis progressed with time. However, no bone formation could be found in the β-TCP scaffolds without the cells at each time point checked. Conclusion Novel β-TCP scaffolds prepared by the polymeric sponge method have excellent osteogenic potentials and can be used as biomaterials for bone tissue engineering.

关 键 词：Β-磷酸三钙 有机泡沫浸渍法 骨髓基质干细胞 成骨能力 

分 类 号：R318.08[医药卫生—生物医学工程] R336[医药卫生—基础医学]