Zhang Zheng, Li Xinyue, Zhao Jing, Jia Wenjun, Wang Zuomin
Department of Periodontology, Tianjin Stomatological Hospital, Hospital of Stomatology, Nankai University, Tianjin, China.
Department of Stomatology, Beijing Chao-Yang Hospital, Capital Medical University, Beijing, China.
PeerJ. 2019 Oct 31;7:e7984. doi: 10.7717/peerj.7984. eCollection 2019.
Platelet concentrates have been used in tissue regeneration. The purpose of this study was to examine effects of growth factors released from leukocyte- and platelet-rich fibrin (L-PRF) and concentrated growth factor (CGF) on the osteogenic differentiation of periodontal ligament fibroblasts (PDLFs).
Leukocyte- and platelet-rich fibrins, CGFs and PDLFs were obtained from New Zealand rabbits. The release of basic fibroblast growth factor (bFGF), bone morphogenetic protein 2 (BMP-2) and transforming growth factor β1 (TGF-β1) from L-PRFs and CGFs was measured at 5 h and 1, 3, 5, 7 days, using the enzyme linked immunosorbent assay. The PDLFs were treated with exudates of L-PRF or CGF. After the treatment, cell counting kit-8 assay was performed at day 1, 3, 5 and 7. Alkaline phosphatase (ALP) assay and Western blotting were applied at day 7. Three blocking antibodies were used to neutralize the proteins of bFGF, BMP-2 and TGF-β1.
Leukocyte- and platelet-rich fibrin and CGF showed different growth factor release pattern, but similar accumulated concentration of these growth factors. PDLFs proliferation was significantly promoted by both L-PRF and CGF at day 1, 3 and 7, and CGF group was superior to L-PRF group at day 1 and 3. Both L-PRF and CGF significantly enhanced PDLFs ALP activity and protein expression of osteogenic markers. The osteopontin level was higher in CGF group than in L-PRF group, but no significant differences were found between two groups for ALP activity. Three blocking antibodies significantly downregulated both L-PRF and CGF induced osteogenic markers expression.
Both CGF and L-PRF can promote the proliferation and osteogenic differentiation of PDLFs. The bFGF, BMP-2 and TGF-β1 are involved in both L-PRF and CGF induced osteogenic differentiation of PDLFs.
血小板浓缩物已用于组织再生。本研究的目的是检测富含白细胞和血小板的纤维蛋白(L-PRF)及浓缩生长因子(CGF)释放的生长因子对牙周膜成纤维细胞(PDLFs)成骨分化的影响。
从新西兰兔获取富含白细胞和血小板的纤维蛋白、CGF及PDLFs。采用酶联免疫吸附测定法在5小时以及第1、3、5、7天测量L-PRF和CGF中碱性成纤维细胞生长因子(bFGF)、骨形态发生蛋白2(BMP-2)和转化生长因子β1(TGF-β1)的释放量。用L-PRF或CGF的渗出液处理PDLFs。处理后,在第1、3、5和7天进行细胞计数试剂盒-8检测。在第7天进行碱性磷酸酶(ALP)检测和蛋白质印迹分析。使用三种阻断抗体中和bFGF、BMP-2和TGF-β1的蛋白质。
富含白细胞和血小板的纤维蛋白和CGF呈现不同的生长因子释放模式,但这些生长因子的累积浓度相似。在第1、3和7天,L-PRF和CGF均显著促进PDLFs增殖,且在第1和3天CGF组优于L-PRF组。L-PRF和CGF均显著增强PDLFs的ALP活性和成骨标志物的蛋白质表达。CGF组骨桥蛋白水平高于L-PRF组,但两组间ALP活性无显著差异。三种阻断抗体均显著下调L-PRF和CGF诱导的成骨标志物表达。
CGF和L-PRF均可促进PDLFs的增殖和成骨分化。bFGF、BMP-2和TGF-β1参与L-PRF和CGF诱导的PDLFs成骨分化过程。