Kakudo Natsuko, Minakata Tatsuya, Mitsui Toshihito, Kushida Satoshi, Notodihardjo Frederik Zefanya, Kusumoto Kenji
Osaka, Japan From the Department of Plastic and Reconstructive Surgery, Kansai Medical University.
Plast Reconstr Surg. 2008 Nov;122(5):1352-1360. doi: 10.1097/PRS.0b013e3181882046.
This study evaluated changes in platelet-derived growth factor (PDGF)-AB and transforming growth factor (TGF)-beta1 release from platelets by platelet-rich plasma activation, and the proliferation potential of activated platelet-rich plasma and platelet-poor plasma on human adipose-derived stem cells and human dermal fibroblasts.
Platelet-rich plasma was prepared using a double-spin method, with the number of platelets counted in each preparation stage. Platelet-rich and platelet-poor plasma were activated with autologous thrombin and calcium chloride, and levels of platelet-released PDGF-AB and TGF-beta1 were determined by enzyme-linked immunosorbent assay. Cells were cultured for 1, 4, or 7 days in serum-free Dulbecco's Modified Eagle Medium supplemented with 5% whole blood plasma, nonactivated platelet-rich plasma, nonactivated platelet-poor plasma, activated platelet-rich plasma, or activated platelet-poor plasma. In parallel, these cells were cultured for 1, 4, or 7 days in serum-free Dulbecco's Modified Eagle Medium supplemented with 1%, 5%, 10%, or 20% activated platelet-rich plasma. The cultured human adipose-derived stem cells and human dermal fibroblasts were assayed for proliferation.
Platelet-rich plasma contained approximately 7.9 times as many platelets as whole blood, and its activation was associated with the release of large amounts of PDGF-AB and TGF-beta1. Adding activated platelet-rich or platelet-poor plasma significantly promoted the proliferation of human adipose-derived stem cells and human dermal fibroblasts. Adding 5% activated platelet-rich plasma to the medium maximally promoted cell proliferation, but activated platelet-rich plasma at 20% did not promote it.
Platelet-rich plasma can enhance the proliferation of human adipose-derived stem cells and human dermal fibroblasts. These results support clinical platelet-rich plasma application for cell-based, soft-tissue engineering and wound healing.
本研究评估了通过富血小板血浆激活,血小板衍生生长因子(PDGF)-AB和转化生长因子(TGF)-β1从血小板中的释放变化,以及激活的富血小板血浆和贫血小板血浆对人脂肪来源干细胞和人真皮成纤维细胞的增殖潜力。
采用双离心法制备富血小板血浆,并在每个制备阶段对血小板数量进行计数。用自体凝血酶和氯化钙激活富血小板血浆和贫血小板血浆,通过酶联免疫吸附测定法测定血小板释放的PDGF-AB和TGF-β1水平。将细胞在补充有5%全血血浆、未激活的富血小板血浆、未激活的贫血小板血浆、激活的富血小板血浆或激活的贫血小板血浆的无血清杜尔贝科改良伊格尔培养基中培养1、4或7天。同时,将这些细胞在补充有1%、5%、10%或20%激活的富血小板血浆的无血清杜尔贝科改良伊格尔培养基中培养1、4或7天。对培养的人脂肪来源干细胞和人真皮成纤维细胞进行增殖测定。
富血小板血浆中的血小板数量约为全血的7.9倍,其激活与大量PDGF-AB和TGF-β1的释放有关。添加激活的富血小板血浆或贫血小板血浆可显著促进人脂肪来源干细胞和人真皮成纤维细胞的增殖。向培养基中添加5%激活的富血小板血浆可最大程度地促进细胞增殖,但20%的激活富血小板血浆则无此促进作用。
富血小板血浆可增强人脂肪来源干细胞和人真皮成纤维细胞的增殖。这些结果支持富血小板血浆在基于细胞的软组织工程和伤口愈合中的临床应用。