富含血小板血浆双相接种法构建组织工程骨(1)
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[摘要]目的：传统的细胞接种法接种效率低，影响组织工程骨的成骨质量。本研究预探讨富含血小板血浆双相接种法是否可以提高细胞接种效率、优化组织工程骨的构建。方法：分别用富含血小板血浆、乏血小板血浆重悬BMSCs，灌注于三维多孔PLGA支架中，凝血酶促凝，体外培养。通过扫描电镜观察支架表面的细胞分布；通过检测支架内DNA含量、ALP含量、成骨基因表达情况分析细胞的接种效率、增殖和分化情况，并与传统的接种方法进行比较。结果：富含血小板血浆和乏血小板血浆双相接种法均能提高细胞与三维支架的结合效率；富含血小板血浆还可以促进支架内细胞的增殖，同时不影响细胞的分化。结论：双相接种法可以提高细胞的接种效率；与普通双相接种法相比，富含血小板血浆双相接种法可以进一步优化组织工程骨的构建。
[关键词]富含血小板血浆；细胞接种；骨组织工程
[中图分类号]R318[文献标识码]A[文章编号]1008-6455(2011)03-0402-06
Constructing tissue engineering bone by a biphasic seeding strategy with platelet-rich plasma
LEI Hua,XIAO Ran,CAO Rui,LV Xiao-yan,GUI Lai
(The Sixth Department,Plastic Surgery Hospital,Peking Union Medical College, China Academy of Medical Sciences,Peking,Beijing 100144,China)
Abstract:ObjectiveThe conventional static seeding is not efficient in constructing tissue engineering bone, which impacts the quality of osteogenesis. This study wants to prove if a biphasic seeding strategy with platelet-rich plasma (PRP) can improve the conventional static seeding.MethodsThe BMSCs suspension in PRP or platelet-poor plasma (PPP) was infiltrated in 3D PLGA scaffolds, clotted by thrombin and cultured for different days. The cell seeding efficiency was evaluated by SEM and by measuring initial DNA contents in 3D scaffolds. The cell proliferation was evaluated by measuring DNA contents in 3D scaffolds in different culture days.The cell differentiation was evaluated by measuring ALP contents and osteogenesis gene expression in 3D scaffolds.Results PRP and PPP could improve the integration of cells and 3D scaffolds. PRP also could enhance the cell proliferation and did not impact the cell differentiation in 3D scaffolds.ConclusionThe biphasic seeding strategy can improve the cell seeding efficiency. PRP can optimize the constructing of tissue engineering bone.
Key words: platelet-rich plasma (PRP);cell seeding;bone tissue engineering
骨组织工程学中，细胞与三位支架的结合即细胞接种是非常重要的一步，目前常用的细胞接种方法仍是静置接种法，即细胞悬液直接滴加在支架表面。此方法适用于各种细胞和支架，但效率低，不仅细胞流失多，细胞也不易均匀布于支架内部[1-2]。因此，有很多研究探讨如何将细胞高效且均匀地分布于支架内。其中有学者将细胞重悬于纤维蛋白原[3-6]或胶原[7-13]的溶液中，将此溶液灌注于支架中，再用促凝剂使溶液凝成凝胶，固定细胞于支架内--即双相接种法。研究表明此方法不仅使细胞与支架的结合效率提高，而且细胞可以灌注于支架内部，有利于细胞的均匀分布。
本研究试用富含血小板血浆(platelet-rich plasma ......
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