背景:添加富血小板血浆可促进细胞体外成骨表型的快速转化,从而有效成骨.目的:观察富血小板血浆对脂肪间充质干细胞体内和体外成骨能力的影响.方法:第3代兔脂肪间充质干细胞进行成骨诱导培养,分为对照组和富血小板血浆组.细胞接种到钙磷陶瓷支架上后,体内外观察细胞/载体复合物的成骨情况.结果与结论:两组细胞随着诱导时间的延长碱性磷酸酶活性增高,达到高峰值后随后逐渐下降,诱导后14 d时,富血小板血浆组即达到高峰值,对照组18 d达到高峰值.细胞/载体复合体切片Von Kossa染色显示两组载体的孔隙内衬面呈多层黑染状,有大量钙盐沉积.甲苯胺蓝染色显示载体的孔隙中可见成熟的骨质存在,周边区域较中心多.体外钙盐沉积对照组多,体内成骨面积富血小板血浆组多(P 〈 0.05).说明,富血小板血浆可有效诱导脂肪间充质干细胞体内和体外成骨.
BACKGROUND: Platelet-rich plasma (PRP) can promote in vitro rapid transformation of osteoblastic phenotype. OBJECTIVE: To investigate the effects of PRP on in vitro and in vivo osteoblastic ability of adipose-derived mesenchymal stem cells (ADSCs). METHODS: ADSCs form rabbits in passage 3 were cultured by osteogenic induction culture medium (control group) or osteogenic induction culture medium containing 10 mL/L PRP. ADSCs of passage 3 were implanted into hydroxyapatite/tricalcium phosphate (HA/TCP) scaffold and cultured by osteogenic induction culture medium with or without 10 mL/L PRP to detect the bone formation. RESULTS AND CONCLUSION: Alkaline phosphatase activity increased to the maximal level on day 14 in the PRP group and on day 18 in the control group, and then gradually declined, but remained a relative high level until to day 28. Histologically Von Kossa staining showed that a lot of mineralized bone nodules which stained black adhered to the wall of inside porous scaffold in two groups. A large number of calcium deposits were found. Toluidine blue dye showed that new bone formed in the pores, which was greater in the surrounding area than in the central area. The amount of in vitro calcium deposits was greater in the control group, and the amount of new bone was more in the PRP group than the control group (P 0.05). PRP could effectively speed up the osteogenic differentiation of ADSCs in vitro and induce bone formation in vivo.