富血小板纤维蛋白对牙周膜成纤维细胞成骨能力影响的实验研究  

Effects of Platelet-rich Fibrin Extract on Periodontal Ligament Cells

作　　者：黄奕智 郭俊[1] 杨健[1] HUANG Yizhi;GUO Jun;YANG Jian(Department of Endodontics,Affiliated Stomatological Hospital of Nanchang University,Nanchang 330000,China)

机构地区：[1]南昌大学附属口腔医院牙体牙髓科江西省口腔生物医学重点实验室

出　　处：《口腔医学研究》2020年第1期41-45,共5页Journal of Oral Science Research

摘　　要：目的:探讨富血小板纤维蛋白提取液(platelet-rich fibrin,PRF)对人牙周膜细胞(human periodontal ligament cells,hPDLCs)增殖、迁移及成骨能力的影响。方法:原代培养hPDLCs,经鉴定后,MTT法检测PRF对hPDLCs增殖的影响;实验分为对照组和PRF组(浓度50%),采用碱性磷酸酶法检测hPDLCs培养7 d和14 d后碱性磷酸酶(alkaline phosp-hatase,ALP)活性;Transwell检测hPDLCs迁移的情况;茜素红染色观察细胞矿化功能;Western Blotting检测骨形态发生蛋白2(bone morphogenetic protein-2,BMP2)和runt相关转录因子2(runt-related transcription factor-2,Runx2)的表达。结果:MTT结果显示,各组A值随着时间延长均逐渐增加,增加值最快的是PFR2组;与对照组相比,在第7 d和14 d时PRF组ALP活性均明显升高(P<0.05);Transwell结果显示,PFR组迁移细胞数较对照组明显增多(P<0.05);茜素红染色结果显示,各时间点PRF组矿化结节A值较对照组更高(P<0.05);Western Blotting结果显示,PFR组中Runx2、BMP2蛋白表达均高于对照组(P<0.05)。结论:PRF具有促进hPDLCs增殖,迁移和成骨分化的作用,其促进骨形成机制需进一步研究。Objective:To investigate the effect of platelet-rich fibrin extract(PRF)on the proliferation,migration,and osteogenesis of human periodontal ligament cells(hPDLCs).Methods:The hPDLCs were primarily cultured.After identification,the effect of PRF on the proliferation of hPDLCs was detected by MTT.The cells was divided into control group and PRF group(concentration 50%).The ALP activity of hPDLCs cultured for 7d and 14d was detected by alkaline phosphatase assay.The migration of hPDLCs was detected by transwell.The mineralization function of cells was observed by alizarin red staining.The expression of BMP2 and Runx2 was detected by western blotting.Results:MTT results showed that OD value of each group increased gradually with time,and the fastest increase was in PFR2 group.Compared with the control group,ALP activity in PRF group increased significantly on day 7 and 14(P<0.05).Transwell results showed that the number of migrating cells in PFR group was significantly higher than that in control group(P<0.05).Alizarin red staining showed that the optical density of mineralized nodules in PRF group was higher than that in control group at all time points(P<0.05).Western blotting results showed that the expressions of Runx2 and BMP2 in PFR group were higher than those in control group(P<0.05).Conclusion:PRF can promote the proliferation,migration,and osteogenic differentiation of hPDLCs,and its mechanism of promoting bone formation needs further study.

关 键 词：富血小板纤维蛋白 细胞增殖 成骨 人牙周膜细胞 

分 类 号：R781.4[医药卫生—口腔医学]