摘要
目的观察并比较不同离心条件制得的富血小板纤维蛋白(PRF)的生长因子释放水平及对成纤维细胞增殖、迁移的作用。方法采用心脏采血法获得SD大鼠全血,分别以708 g 12 min、208 g 14 min、208 g 8 min三种离心程序获得富白细胞PRF(L-PRF)、高级PRF(A-PRF)、高级PRF+(A-PRF+),压制得到PRF膜片后,置于DMEM/F12培养基中,分别于1 h、6 h、1 d、3 d、5 d后收集培养基,采用ELISA法检测培养基中的血小板衍生生长因子(PDGF-AB)、转化生长因子β(TGF-β)、血管内皮生长因子(VEGF)。取SD大鼠背部皮肤分离、培养获得皮肤成纤维细胞,将细胞分为L-PRF组、A-PRF组、A-PRF+组、空白组,分别以L-PRF、A-PRF、A-PRF+条件培养基和10%FBS培养基培养,采用CCK-8细胞增殖实验检测细胞增殖能力、Transwell小室实验检测细胞迁移能力。结果制备5 d内A-PRF、A-PRF+培养基的PDGF-AB、TGF-β、VEGF总释放水平高于L-PRF培养基(P均<0.05)。A-PRF+组、A-PRF组、L-PRF组培养3、5 d时细胞增殖能力均高于空白组,且培养3 d时A-PRF+组、A-PRF组细胞增殖能力高于L-PRF组(P均<0.05)。L-PRF组、A-PRF组、A-PRF+组穿膜细胞数高于空白组,且A-PRF组、A-PRF+组穿膜细胞数高于L-PRF组(P均<0.05)。结论通过降低相对离心力制备得到的A-PRF+和A-PRF的生长因子释放性能、促进成纤维细胞增殖和迁移作用明显强于L-PRF,且适当缩短离心时间得到的A-PRF+作用更强。
Objective To observe and compare the growth factor release levels of platelet-rich fibrin(PRF)prepared under different centrifugation conditions and its effect on the proliferation and migration of fibroblasts.Methods The whole blood of SD rats was obtained by cardiac blood sampling,and the leukocyte-rich PRF(L-PRF),advanced PRF(A-PRF),and advanced PRF+(A-PRF+)were obtained by three centrifugal procedures of 708 g 12 min,208 g 14 min,and 208 g 8 min.The PRF diaphragm was pressed and placed in DMEM/F12 medium which was then collected at 1 h,6 h,1 d,3 d,and 5 d,respectively.The platelet-derived growth factor(PDGF-Ab),transforming growth factorβ(TGF-β),and vascular endothelial growth factor(VEGF)in the culture medium were detected by ELISA.Skin fibroblasts were isolated and cultured from the dorsal skin of SD rats,and then were divided into the L-PRF group,A-PRF group,A-PRF+group,and blank group,which were cultured in L-PRF,A-PRF,and A-PRF+conditioned medium and 10%FBS medium,respectively.CCK-8 cell proliferation assay was used to detect cell proliferation,and Transwell chamber assay was used to detect cell migration.Results The overall release levels of PDGF-Ab,TGF-βand VEGF in A-PRF and A-PRF+medium were higher than those in L-PRF medium within 5 days after preparation(all P<0.05).The cell proliferation abilities of the A-PRF+group,A-PRF group,and L-PRF group were higher than that of blank group on day 3 and 5,and the cell proliferation abilities of the A-PRF+group and A-PRF group were higher than that of L-PRF group on day 3(all P<0.05).The numbers of transmembrane cells in the L-PRF group,A-PRF group,and A-PRF+group were higher than that in the blank group,and the numbers of transmembrane cells in the A-PRF group and A-PRF+group were higher than that in the L-PRF group(all P<0.05).Conclusion The effects of A-PRF+and A-PRF prepared by reducing the relative cen⁃trifugal force on the growth factor release and promoting the proliferation and migration of fibroblasts were significantly stron⁃ger than that of L
作者
贺波
霍继武
熊竹友
蒋邦红
陈宇
李光早
HE Bo;HUO Jiwu;XIONG Zhuyou;JIANG Banghong;CHEN Yu;LI Guangzao(The First Affiliated Hospital of Bengbu Medical College,Bengbu 233000,China;不详)
出处
《山东医药》
CAS
2021年第8期24-27,共4页
Shandong Medical Journal
基金
安徽省科技攻关项目(1704a0802162)
安徽省教育厅重点项目(KJ2019A0345)。
