β-磷酸三钙复合成骨细胞的异位成骨

Ectopic osteogenesis of osteoblasts combined with beta-tricalcium phosphate

背景:β-磷酸三钙作为理想的骨替代材料己成为众多学者研究的重点之一,但与新生大鼠颅盖骨来源成骨细胞复合的研究较少。目的:评价多孔β-磷酸三钙支架与成骨细胞复合植入大鼠肌袋模型后的异位成骨能力。设计、时间及地点:同体对照动物实验,于2007-09/2008-10在西安交通大学医学院中心实验室和实验医学动物中心完成。材料:新生SD大鼠2只,12周龄SD大鼠20只。多孔β-磷酸三钙购自上海贝奥路公司,大小为5mm×5mm×5mm。方法:应用扫描电镜观察β-磷酸三钙结构特点。成骨细胞体外分离、培养,倒置显微镜下观察细胞生长情况。使用第3代细胞与β-磷酸三钙复合后,植入大鼠背部左侧肌袋,将β-磷酸三钙空白支架植入右侧肌袋。分别在植入1,4,8周后,将成骨细胞β-磷酸三钙复合物取出行苏木精-伊红染色观察新骨形成情况,并应用KS4003.0软件进行新骨生成分析。主要观察指标:①β-磷酸三钙结构特征。②成骨细胞生长情况。③组织学观察新骨生成情况。④图像分析新骨生成结果。结果:β-磷酸三钙孔径大小一般为(500±150)μm,孔内连接径(150±50)μm。倒置显微镜下观察成骨细胞呈三角形、多角形或梭形。植入1周后所有标本都未发现骨生成;在4,8周时成骨细胞复合β-磷酸三钙组新骨生成量均多于β-磷酸三钙组(P<0.05,0.01)。结论:多孔β-磷酸三钙复合成骨细胞支架在大鼠体内具有异位成骨能力。

BACKGROUND： As an ideal replace materials, β-tricalcium phosphate （β -TCP） has become one of the key researches, however, studies regarding composites of β -TCP and osteoblasts in neonatal calvaria cultures are few. OBJECTIVE： To evaluate the ability of ectopic osteogenesis of composites osteoblasts with β -TCP in rat＇s muscle pouch. DESIGN, TIME AND SETTING： The homebody control experiment was performed at the Central Laboratory and Animal Experimental Center of Xi＇an Jiaotong University from September 2007 to October 2008. MATERIALS： Two new born Sprague Dawley rats, and 20 rats with 12-weeks-old. β-TCP, with size of 5 mm×5 mm×5 mm, was purchased from Shanghai Bio-lu Biomaterials Co., Ltd. METHODS： Structural feature of.β -TCP was observed under scanning electron microscope. Isolated and cultured osteoblasts in vitro, growth status of cells was observed by inverted microscope. The third passage of osteoblasts was combined with β -TCP, and implanted into muscle pouch of left side, the β -TCP was implanted into right side. Specimens were harvested at weeks 1, 4 and 8 and analyzed by hematoxylin-eosin staining. The percentages of new bone formation were determined by KS400 3.0 software. MAIN OUTCOME MEASURES： （1）The structural feature of β -tricalcium phosphate. （2）Growth status of osteoblasts. （3） Osteogenic capability was observed by histological observation. （4）Image analysis the percentages of new bone formation. RESULTS： The spherical pores diamete of β-TCP was （500±150） μ m, with （150±50） μ m interconnections in diameter. Osteoblasts were showed triangle, polygon and fusiform shape under inverted microscope. After 1 week of implantation, there was no new bone formation in all groups. At weeks 4 and 8, the amount of bone formation in the osteoblasts/β -TCP group was greater than that of β -TCP group（P 〈 0.05, 0.01）. CONCLUSION： Bone substitutes constructed byporous β-TCP combined with osteoblasts have the ability ofectopic osteogenesis.