富血小板纤维蛋白对牙龈成纤维细胞生物学功能的影响

doi:10.13701/j.cnki.kqyxyj.2024.07.008

摘要/Abstract

摘要： 目的:探究冷冻干燥富血小板纤维蛋白(freeze-dried platelet-rich fibrin,FD-PRF)对人牙龈成纤维细胞(human gingival fibroblasts,hGFs)功能的影响以及其抗菌性能,为口腔黏膜炎的防治提供新思路。方法:采用流式细胞术、CCK-8检测细胞增殖(cell counting kit-8,CCK-8)、细胞划痕和Transwell实验,检测FD-PRF对hGFs细胞周期、增殖和迁移的影响;采用倾注平板法和振荡培养法研究FD-PRF的抗菌作用。结果:CCK-8实验结果表明,随着FD-PRF浓度升高,hGFs的细胞数量显著增加。细胞周期结果显示, S-G2/M期的细胞比例:5%FD-PRF组>1%FD-PRF组>对照组,而G0-G1期细胞则相反。迁移实验结果显示,FD-PRF显著提升了hGFs的迁移速度。抗菌实验结果显示,FD-PRF组相比于对照组的大肠杆菌和金黄色葡萄球菌数量明显减少。结论:FD-PRF促进hGFs进入细胞分裂期,进而促进细胞增殖。FD-PRF促进hGFs迁移。此外,本研究还证实FD-PRF具有一定的抗菌性。提示FD-PRF可以促进口腔黏膜炎的愈合。

关键词: 冷冻干燥富血小板纤维蛋白, 成纤维细胞, 抗菌性, 口腔黏膜炎

Abstract: Objective: To explore the effects of freeze-dried platelet-rich fibrin (FD-PRF) on human gingival fibroblasts (hGFs) and antibacterial property of FD-PRF, and to provide a new way for controlling refractory oral mucositis (OM). Methods: Cell counting kit-8 (CCK-8), flow cytometry, scratch wound, and Transwell assays were used to investigate the effects of FD-PRF on hGFs’ cycle, proliferation, and migration. Pour plate and shaking culture methods were explored to investigate the antibacterial ability of FD-PRF. Results: CCK-8 assay showed that hGFs number increased significantly with the increase of FD-PRF concentration. In cell cycle results, proportions of cells at S-G2/M phase: 5%FD-PRF group>1%FD-PRF group > control group. And proportions of cells at G1-G0 phase were the opposite. Cells migration assays suggested that FD-PRF improved migration rate of hGFs. For antibacterial function tests of FD-PRF, it was found that clones of E. coli and S. aureus were both reduced in the FD-PRF group. Conclusion: By facilitating hGFs entering the division stage, FD-PRF promotes proliferation of hGFs. FD-PRF possesses significant chemotaxis toward hGFs. Besides, FD-PRF possesses intrinsic antibacterial property. FD-PRF may promote the healing of oral mucositis.

Key words: freeze-dried platelet-rich fibrin, fibroblasts, antibacterial property, oral mucositis

王玉花, 柴纪华, 张好建. 富血小板纤维蛋白对牙龈成纤维细胞生物学功能的影响[J]. 口腔医学研究, 2024, 40(7): 611-616.

WANG Yuhua, CHAI Jihua, ZHANG Haojian. Effects of Platelet-rich Fibrin on Biological Function of Gingival Fibroblasts[J]. Journal of Oral Science Research, 2024, 40(7): 611-616.

参考文献

[1] Abdulrahman YA, Hosny MM, Elfana A, et.al. Clinical and radiographic evaluation of low-speed platelet-rich fibrin (PRF) for the treatment of intra-osseous defects of stage-Ⅲ periodontitis patients: a randomized controlled clinical trial [J]. Clin Oral Investig, 2022, 26(11):6671-6680.
[2] Beaudoin PL, Carles G. Platelet-rich fibrin in rhinoplasty: A precise and standardized approach [J]. Eur Ann Otorhinolaryngol Head Neck Dis, 2023, 140(6):317-321.
[3] Clark D, Rajendran Y, Paydar S, et al. Advanced platelet-rich fibrin and freeze-dried bone allograft for ridge preservation: A randomized controlled clinical trial [J]. J Periodontol, 2018, 89(4):379-387.
[4] Miron RJ, Gruber R, Farshidfar N, et al. Ten years of injectable platelet-rich fibrin [J]. Periodontol 2000, 2024, 94(1):92-113.
[5] El Bagdadi K, Kubesch A, Yu X, et al. Reduction of relative centrifugal forces increases growth factor release within solid platelet-rich-fibrin (PRF)-based matrices: a proof of concept of LSCC (low speed centrifugation concept) [J]. Eur J Trauma Emerg Surg, 2019, 45(3):467-479.
[6] Ghanaati S, Booms P, Orlowska A, et al. Advanced platelet-rich fibrin: a new concept for cell-based tissue engineering by means of inflammatory cells [J]. J Oral Implantol, 2014, 40(6):679-689.
[7] Nakatani Y, Agata H, Sumita Y, et al. Efficacy of freeze-dried platelet-rich plasma in bone engineering [J]. Arch Oral Biol, 2017, 73:172-178.
[8] Shiga Y, Orita S, Kubota G, et al. Freeze-dried platelet-rich plasma accelerates bone union with adequate rigidity in posterolateral lumbar fusion surgery model in rats [J]. Sci Rep, 2016, 6:36715.
[9] Ngah NA, Dias GJ, Tong DC, et al. Lyophilised platelet-rich fibrin: Physical and biological characterisation [J]. Molecules, 2021, 26(23):7131.
[10] Zhang J, Qi X, Luo X, et al. Clinical and immunohistochemical performance of lyophilized platelet-rich fibrin (Ly-PRF) on tissue regeneration [J]. Clin Implant Dent Relat Res, 2017, 19(3):466-477.
[11] Wang Z, Han L, Sun T, et al. Preparation and effect of lyophilized platelet-rich fibrin on the osteogenic potential of bone marrow mesenchymal stem cells in vitro and in vivo [J]. Heliyon, 2019, 5(10):e02739.
[12] Liu Z, Jin H, Xie Q, et al. Controlled release strategies for the combination of fresh and lyophilized platelet-rich fibrin on bone tissue regeneration [J]. Biomed Res Int, 2019, 2019:4923767.
[13] Azmi AF, Yahya M, Azhar NA, et al. In vitro cell proliferation and migration properties of oral mucosal fibroblasts: A comparative study on the effects of cord blood- and peripheral blood-platelet lysate [J]. Int J Mol Sci, 2023, 24(6):5775.
[14] Waasdorp M, Krom BP, Bikker FJ, et al. The bigger picture: Why oral mucosa heals better than skin [J]. Biomolecules, 2021, 11(8):1165.
[15] Attinà G, Romano A, Maurizi P, et al. Management of oral mucositis in children with malignant solid tumors [J]. Front Oncol, 2021, 11:599243.
[16] Elad S, Yarom N, Zadik Y, et al. The broadening scope of oral mucositis and oral ulcerative mucosal toxicities of anticancer therapies [J]. CA Cancer J Clin, 2022, 72(1):57-77.
[17] Lorini L, Perri F, Vecchio S, et al. Confounding factors in the assessment of oral mucositis in head and neck cancer [J]. Support Care Cancer, 2022, 30(10):8455-8463.
[18] Nishii M, Soutome S, Kawakita A, et al. Factors associated with severe oral mucositis and candidiasis in patients undergoing radiotherapy for oral and oropharyngeal carcinomas: a retrospective multicenter study of 326 patients [J]. Support Care Cancer, 2020, 28(3):1069-1075.
[19] Alfonso García SL, Parada-Sanchez MT, Arboleda Toro D. The phenotype of gingival fibroblasts and their potential use in advanced therapies [J]. Eur J Cell Biol, 2020, 99(7):151123.
[20] Lohana P, Suryaprawira A, Woods EL, et al. Role of enzymic antioxidants in mediating oxidative stress and contrasting wound healing capabilities in oral mucosal/skin fibroblasts and tissues [J]. Antioxidants (Basel), 2023, 12(7):1374.
[21] Dally J, Khan JS, Voisey A, et al. Hepatocyte growth factor mediates enhanced wound healing responses and resistance to transforming growth factor-β₁-driven myofibroblast differentiation in oral mucosal fibroblasts [J]. Int J Mol Sci, 2017, 18(9):1843.
[22] Feng M, Wang Y, Zhang P, et al. Antibacterial effects of platelet-rich fibrin produced by horizontal centrifugation [J]. Int J Oral Sci, 2020, 12(1):32.
[23] Xu F, Zou D, Dai T, et al. Effects of incorporation of granule-lyophilised platelet-rich fibrin into polyvinyl alcohol hydrogel on wound healing [J]. Sci Rep, 2018, 8(1):14042.
[24] Straub A, Utz C, Stapf M, et al. Impact of aminopenicillin administration routes on antimicrobial effects of platelet-rich fibrin: An in-vitro investigation [J]. J Stomatol Oral Maxillofac Surg, 2023, 125(3):101725.