【摘要】 背景:使用富血小板血浆(PRP)等凝胶样物质作为接种介质的双相接种法可优化组织工程骨构建,但PRP的促成骨作用仍存在较大争议。目的:进一步探讨自体PRP凝胶用于双相接种法构建组织工程骨的可行性,及其促成骨性能。方法:实验组利用自体PRP凝胶接种骨髓间充质干细胞(MSCs)于β-磷酸三钙(β-TCP)支架构建组织工程骨,对照组采用常规的DMEM静态接种法构建。体外实验:通过MTT实验、碱性磷酸酶(ALP)活性检测、骨钙素(OC)放免测定,比较两种方法对细胞增殖和成骨分化影响。体内实验:将构建材料植入兔桡骨大段骨缺损,通过组织学观察、X线检查、单光子发射计算机断层检查比较两种方法构建材料的成骨性能。结果:体外实验表明PRP组MSCs显示良好的增殖和成骨分化能力,明显优于对照组。体内实验表明PRP组修复大段骨缺损的能力显著优于对照组。结论:自体PRP具有良好的促成骨性能,可有效构建组织工程骨。更多还原

【Abstract】 Background: The platelet-rich plasma(PRP) gels used in the biphasic seeding can optimize the construction of tissue-engineered bone. However, osteogenic capacity of PRP is still in dispute. Objective: To explore the feasibility of constructing tissue-engineered bone by biphasic seeding with autologous PRP gels so as to give more insight to the contradictory osteogenic capacity of PRP. Methods: Autologous PRP gel was applied to inoculate mesenchymal stromal cells(MSCs)into β-tricalcium phosphate(β-TCP) scaffolds to construct tissue engineered bone in PRP group. In control group, tissue engineered bone was established by conventional static seeding using Dulbecco’s modified Eagle’s medium(DMEM) as cell seeding vehicle. In vitro, proliferation and osteogenic differentiation of MSCs on fabricated constructs were investigated by MTT assay, alkaline phosphatase(ALP) activity and osteocalcin(OC) content. The in vivo bone-forming capacity of the fabricated constructs in rabbit radius was determined by radiography, single photon emission computed tomography and histological analysis. Results: The MSCs in PRP group demonstrated great cell proliferation and osteogenic differentiation in vitro and superior bone formation in vivo as compared with control group. Conclusions: Autologous PRP has a good osteogenic capacity. The biphasic seeding with the use of autologous PRP gel can effectively reconstruct tissue-engineered bones.更多还原