Platelet-rich plasma/osteoblasts complex induces bone formation via osteoblastic differentiation following subcutaneous transplantation
Platelet-rich plasma/osteoblasts complex induces bone formation via osteoblastic differentiation following subcutaneous transplantation
复制标题
DOI：
10.1111/j.1600-0765.2006.00892.x
复制
发表时间：
2006-10-01
影响因子：
3.5
通讯作者：
Izumi, Y.
中科院分区：
文献类型：
--
作者：
Goto, H.;Matsuyama, T.;Izumi, Y.
背景和目的：富含血小板血浆(PRP)可促进局部应用后的骨移植的成熟，并对体外培养的成骨细胞具有生物学效应。然而，PRP在临床上并没有单独应用，因为它在治疗大型骨缺损方面的益处很差。本研究旨在开发一种临床替代自体骨的方法，通过研究PRP与成骨细胞的联合应用并评价其移植后的效果。材料和方法：从ddy小鼠的血液中通过两步离心法制备PRP和血小板贫乏血浆(PPP)。移植前用荧光碳菁标记MC3T3-E1细胞。将标记细胞和PRP凝胶联合移植到严重联合免疫缺陷(SCID)小鼠背部，4wk后进行放射学和免疫组织学评价。移植前用碱性磷酸酶(ALP)染色和von Kossa染色评价PRP的作用，用逆转录聚合酶链式反应(RT-PCR)分析骨相关标志物的表达。结果：与PPP相比，PRP移植前Osterix和骨涎蛋白mRNAs的表达增强。此外，PRP还能提高ALP活性，诱导矿化结节的形成。移植后，标记细胞与PRP凝胶结合形成矿化组织，荧光显微镜下可见移植细胞表达骨钙素和I型胶原。结论：PRP/成骨细胞复合体的应用具有促进成骨细胞分化的作用。
Background and Objectives: Platelet-rich plasma (PRP) has been shown to enhance the maturation of bone grafts following local application and to have biological effects on osteoblasts in vitro. However, PRP is not applied by itself clinically as a result of its poor benefits in large bone defects. The present study was undertaken to develop a clinical alternative to autologous bone, by investigating the application of PRP in combination with osteoblastic cells and evaluating its effects after transplantation.Material and Methods: PRP and platelet-poor plasma (PPP) were prepared from blood, obtained from ddY mice, by two centrifugation steps. MC3T3-E1 cells were labeled with fluorescent carbocyanine just before transplantation. The combination of labeled cells and PRP gel was subcutaneously transplanted into the back of severe combined immunodeficient (SCID) mice, and the transplants were evaluated radiographically and immunohistologically after 4 wk. The effects of PRP were assessed by alkaline phosphatase (ALP) staining and von Kossa staining, and the expression of bone-related markers was analyzed by reverse transcription-polymerase chain reaction before transplantation.Results: Before transplantation, PRP enhanced the expression of Osterix and bone sialoprotein mRNAs compared with PPP. Furthermore, PRP elevated ALP activity and induced the formation of mineralized nodules. After transplantation, the combination of labeled cells and PRP gel formed mineralized tissue, and the transplanted cells visualized in the tissue using fluorescence microscopy expressed osteocalcin and type I collagen.Conclusion: These results suggest that the application of a PRP/osteoblasts complex has beneficial effects for transplanting engineered cells into bone defects through the promotion of osteoblastic differentiation.