目的:本研究旨在开发一种复合生物材料,以实现有效的软组织再生。
方法:将化合物透明质酸(CHA)和液体水平富血小板纤维蛋白(H-PRF)以1:1的比例混合以形成CHA-PRF凝胶。在本研究中使用人牙龈成纤维细胞(HGF)。CHA的影响,H-PRF,通过CCK-8测定评价CHA-PRF凝胶对细胞活力的影响。然后,CHA的影响,H-PRF,并通过qRT-PCR和免疫荧光分析评价CHA-PRF凝胶对胶原蛋白形成和沉积的影响。最后,qRT-PCR,免疫荧光分析,Transwell分析,和划痕伤口愈合试验进行,以确定CHA,H-PRF,CHA-PRF凝胶影响HGFs的迁移。
结果:CHA和H-PRF的组合缩短了液体H-PRF的凝固时间。与单纯CHA和H-PRF组相比,CHA-PRF组在所有时间点表现出最高的细胞增殖,如CCK-8测定所示。Col1a和FAK在CHA-PRF组中的表达水平最高,如qRT-PCR所示。CHA和PRF可以刺激胶原形成和HGF迁移,通过荧光显微镜分析COL1和F-肌动蛋白以及Transwell和划痕愈合测定观察到。
结论:CHA-PRF组表现出更大的潜力,通过诱导细胞增殖来促进软组织再生,胶原蛋白合成,在HGFs中的迁移比纯CHA或H-PRF组。CHA-PRF可以作为单独使用或与自体移植物组合用于牙周或种植体周围软组织再生的绝佳候选物。
OBJECTIVE: This study aims to develop a compound biomaterial to achieve effective soft tissue regeneration.
METHODS: Compound hyaluronic acid (CHA) and liquid horizontal-platelet-rich fibrin (H-PRF) were mixed at a ratio of 1:1 to form a CHA-PRF gel. Human gingival fibroblasts (HGFs) were used in this study. The effect of CHA, H-PRF, and the CHA-PRF gel on cell viability was evaluated by CCK-8 assays. Then, the effect of CHA, H-PRF, and the CHA-PRF gel on collagen formation and deposition was evaluated by qRT‒PCR and immunofluorescence analysis. Finally, qRT‒PCR, immunofluorescence analysis, Transwell assays, and scratch wound-healing assays were performed to determine how CHA, H-PRF, and the CHA-PRF gel affect the migration of HGFs.
RESULTS: The combination of CHA and H-PRF shortened the coagulation time of liquid H-PRF. Compared to the pure CHA and H-PRF group, the CHA-PRF group exhibited the highest cell proliferation at all time points, as shown by the CCK-8 assay. Col1a and FAK were expressed at the highest levels in the CHA-PRF group, as shown by qRT‒PCR. CHA and PRF could stimulate collagen formation and HGF migration, as observed by fluorescence microscopy analysis of COL1 and F-actin and Transwell and scratch healing assays.
CONCLUSIONS: The CHA-PRF group exhibited greater potential to promote soft tissue regeneration by inducing cell proliferation, collagen synthesis, and migration in HGFs than the pure CHA or H-PRF group. CHA-PRF can serve as a great candidate for use alone or in combination with autografts in periodontal or peri-implant soft tissue regeneration.