仿生合成梯度含锶磷酸钙骨水泥的体外细胞生物学性能检测

Biomimetic synthesis of gradient Sr-containing calcium phosphate cement and cytobiological performance assay in vitro

目的:通过在仿生理条件下构建梯度含锶磷酸钙骨水泥,并应用L-929成纤维细胞和幼兔成骨细胞检测含锶磷酸钙骨水泥的体外生物学性能。方法:实验于2006-11/2007-04在解放军第四军医大学口腔医院颌面外科和西安交通大学金属材料强度国家重点实验室完成。①实验材料:仿生理条件下合成梯度含锶磷酸钙骨水泥。②实验方法:应用了L-929成纤维细胞和成骨细胞两种细胞,将不同浓度的梯度含锶磷酸钙骨水泥分别与两种细胞接触。③评估指标:采用MTT检测法,对细胞增殖活性进行检测,计算细胞相对增殖率,用6级毒性分类法评级;并应用相差显微镜、扫描电镜进行形态学观察。用碱性磷酸酶测定试剂盒检测浸提液对兔成骨细胞功能表达的影响;检测其细胞毒性、促成骨细胞黏附、增殖及表达能力以及生物降解潜能。结果:仿生理条件下合成的梯度含锶磷酸钙骨水泥无细胞毒性,表面成骨细胞的黏附性良好,增殖活跃,并显示出可能存在良好的生物降解能力。结论:含锶磷酸钙骨水泥体外细胞生物相容性良好,是安全的新型骨组织工程支架材料。

AIM： This study constructed novel gradient Sr-containing calcium phosphate cement （CPC） by means of biomimetic synthesis and investigated the biocompatibility of this material using L-929 fibroblasts and young rabbit osteoblasts. METHODS： From November 2006 to April 2007, the experiment was carried out in the Department of Oral-Maxillofacial Surgery, Stomatological College at the Fourth Military Medical University of Chinese PLA and the State Key Laboratory tbr Mechanical Behavior of Materials at Xi＇an Jiaotong University.（1）Novel gradient Sr-containing CPCs were biomimetic synthesized（22）The new biomaterials were contacted with L-929 fibroblast cells and rabbit＇s osteoblasts in different situations. （3）MTT assay was used to detect the cell proliferation activity, and relative proliferation rate was calculated and graded according to 6-level toxicity classification. Morphology of cells was determined by using contrast phase microscope and scanning electron microscope. Alkaline phosphatase assay was applied for the influence of leaching liquor on the rabbit osleoblast function. Cytotoxicity, capacity of protnoting the adhesion, prolifizration and expression of osteoblasts as well as biodegradable potential were all detected. RESULTS： No cytotoxicity, good adhesion and proliferation of osteoblasts to the surface were detected in Sr-containing CPC by means of biomimetic synthesis, indicating active biodegradation might occur in Sr-containing CPC. CONCLUSION： The gradient Sr-containing CPC synthesized biomimetically shows good biocompatibility in vitro, and can bc used safely in tissue engineering bone scaffold.