改良型与可注射型富血小板纤维蛋白对人牙龈成纤维细胞增殖及分化的影响

Effect of advanced platelet-rich fibrin and injectable platelet-rich fibrin on the biological properties of human gingival fibroblasts

目的 探究改良型富血小板纤维蛋白(advanced platelet-rich fibrin, A-PRF)和可注射型富血小板纤维蛋白(injectable platelet-rich fibrin, i-PRF)对人牙龈成纤维细胞增殖及分化的影响,为两种血小板纤维蛋白制品促进软组织愈合的表观差异及临床效能评估提供实验数据参考。方法 观察A-PRF及iPRF显微及超微结构,测定血小板浓缩物生长因子(PDGF-BB,TGF-β1,EGF)的释放情况,将A-PRF及i-PRF作用于人牙龈成纤维细胞(human gingival fibroblasts, HGFs),观察两者对HGFs的增殖影响及HGFs I型胶原蛋白(COL1)的表达情况,通过qRT-PCR进一步检测I型胶原蛋白α2(COL1α2)、纤连蛋白I (FN1)、转移生长因子β(TGF-β)、血小板衍生生长因子(PDGF)基因的表达。结果 i-PRF中白细胞层较A-PRF更宽,且白细胞数量更多,而A-PRF的纤维网状结构更为致密。A-PRF、i-PRF中的细胞因子随着时间的延长,在各时间点释放量逐渐降低,A-PRF中TGF-β1在第1天及EGF在第7天释放量低于i-PRF,其余各个时间点A-PRF生长因子的释放量均高于i-PRF (P<0.05), A-PRF及i-PRF均对HGFs的增殖具有显著促进作用,差异无统计学意义。A-PRF对TGF-β、PDGF基因的上调作用更加明显,而i-PRF对HGFs I型胶原蛋白及COL1a2、FN1基因的上调作用更为明显。结论 A-PRF、i-PRF均对HGFs的增殖具有明显的促进作用,且二者对HGFs I型胶原蛋白的表达起到了显著促进作用,在同一时间段A-PRF可分泌更多的软组织愈合相关生长因子,提示A-PRF在促进软组织修复愈合方面表现更优。

Objective To investigate the effects of advanced platelet-rich fibrin(A-PRF) and injectable platelet-rich fibrin(i-PRF) on the proliferation and differentiation of human gingival fibroblasts. Methods The microstructure and ultrastructure of A-PRF and iPRF were observed and the release of platelet concentrate growth factors(PDGF-BB,TGF-β1, EGF) was determined. A-PRF and i-PRF were applied to human gingival fibroblasts(human gingival fibroblasts,HGFs). The effects of A-PRF and iPRF on the proliferation of HGFs and the expression of HGFs type I collagen(COL1)were observed, and type I collagen α2(COL1α2) and fibronectin I(FN1), transfer growth factor β(TGF-β)and platelet-derived growth factor(PDGF) gene expression were further detected by qRT-PCR. Results The white blood cell layer in i-PRF was wider than that in A-PRF, and the number of white blood cells was larger. The cytokines in A-PRF and i-PRF increased with time. The release volume of TGF-β1 in A-PRF on the 1st day and EGF release on the 7th day were lower than that of i-PRF, and the release volume of A-PRF growth factor at each time point was higher than that of i-PRF(P<0.05).A-PRF and i-PRF both had a significant promotion effect on the proliferation of HGFs. A-PRF had a significant effect on TGF-β and PDGF genes. The up-regulation effect of i-PRF on HGFs type I collagen,COL1a2 and FN1 genes was more obvious. Conclusions Both A-PRF and i-PRF can significantly promote the proliferation of HGFs and the expression of HGFs type I collagen. A-PRF is better in promoting soft tissue repair and healing.