复合骨髓间充质干细胞和富血小板血浆煅烧骨在大鼠下颌骨缺损修复中的应用

Application of calcined bone compounded with bone marrow mesenchymal stem cells and platelet-rich plasma in repairing mandibular defect in rats

背景:用于修复骨缺损的方法有直接植入、复合细胞植入以及加入生长因子植入。近年来,复合细胞植入和加入生长因子植入的研究受到比较广泛的关注。目的:观察复合骨髓间充质干细胞和富血小板血浆的煅烧骨修复颌骨缺损的作用。设计、时间及地点:随机对照动物实验,于2008-04在青岛大学医学院附属医院中心实验室及口腔研究室完成。材料:取成年牛长骨骨骺端,经脱脂、脱蛋白、煅烧处理后制得煅烧骨。将第3代的骨髓间充质干细胞悬液加到煅烧骨上制备煅烧骨-骨髓间充质干细胞复合体。将富血小板血浆因子滴加到将要植入的煅烧骨-骨髓间充质干细胞复合体上,制得含富血小板血浆因子的煅烧骨-骨髓间充质干细胞复合体。方法:用牙钻在Wistar大鼠左侧下颌骨下缘备出1.0cm×0.8cm大小缺损,将30只大鼠随机分为3组,每组10只。对照组:将煅烧骨直接植入大鼠下颌骨缺损中。复合骨髓间充质干细胞组:将煅烧骨-骨髓间充质干细胞复合体植入大鼠下颌骨缺损中。复合骨髓间充质干细胞+富血小板血浆组:将含富血小板血浆因子的煅烧骨-骨髓间充质干细胞复合体植入大鼠下颌骨缺损中。主要观察指标:观察各组煅烧骨在4,8,12周的成骨情况。结果:复合骨髓间充质干细胞+富血小板血浆组煅烧骨在4,8,12周3个时间点的成骨情况明显优于对照组煅烧骨及复合骨髓间充质干细胞组煅烧骨(P<0.01),对照组和复合骨髓间充质干细胞组之间的成骨情况差异无显著性意义(P>0.05)。结论:复合骨髓间充质干细胞和富血小板血浆的煅烧骨可以促进下颌骨缺损修复,增加血管化与骨化。

BACKGROUND： Bone defect usually repaired by direct embedding, compound cells embedding, as well as compound growth factors embedding. Recently, research regarding compound cells embedded calcined bone combined with growth factor were given close attention. OBJECTIVE： To observe the effects of calcined bone modified by bone marrow mesenchymal stem cells （BMSCs） and platelet-rich plasma （PRP） in repairing mandibular defect. DESIGN, TIME AND SETTING： A randomized control experiment was performed at the central laboratory and Department of Stomatology, the Affiliated Hospital of Medical College, Qingdao University in April 2008. MATERIALS： The epiphyseal end of bovine long bone was treated by degrease, deproteinization as well as calcinations to obtain calcined bone. The third passages of BMSCs were added into the calcined bone to prepare calcined bone compounded with BMSCs. Based on this, PRP was dropped to harvest calcined bone compounded with BMSCs and PRP. METHODS： Defects with size of 1.0 cm×0.8 cm was prepared from inferior border of mandible of rats with toothdrill, then randomly divided rats into 3 groups, with 10 animals in each group. Calcined bone, BMSCs combined calcined bone, as well as calcined bone compounded with BMSCs and PRP were respectively implanted in the model of mandibular defects on rats. MAIN OUTCOME MEASURES： The condition of osteogenesis was observed at the weeks 4, 8 and 12 after operation. RESULTS： Bone formation of BMSCs＋PRP group was significant superior to control and BMSCs groups at the weeks 4, 8 and 12 after operation （P 〈 0.01）. However, there was no significant difference between control group and BMSCs group at the weeks 4, 8 and 12 after operation week （P 〉 0.05）. CONCLUSION： The calcined bone added PRP combined with BMSCs can accelerate repairing of mandibular defect, as well as increase vascularization and calcification.