Effect of A-PRF on the biological performance of human bone marrow mesenchymal cells

doi:10.12337/zgkqzzxzz.2021.10.003

Abstract

Abstract: Objective The aim of this study was to investigate the cytobiological effects of advanced-platelet rich fibrin (A-PRF) on cells from human bone marrow mesenchyme (hBMMCs) in vitro. Methods hBMMCs were identified as adipogenic and osteogenic differentiation. A-PRF was observed under a scanning electron microscope and its growth factor was detected by human growth factor antibody array-membrane. CCK-8 and scratch tests were used to detect the effect of A-PRF on the proliferation and migration of hBMMCs at different concentrations. Alkaline phosphatase activity and alizarin red staining were used to detect the effect of different concentrations of A-PRF on the mineralization ability of hBMMCs. Results SEM showed that A-PRF is a three-dimensional network containing more than 40 cytokines, among which macrophage colony stimulating factor and human nerve growth factor had not been reported. 10% A-PRF increased cell proliferation and migration ability (P<0.05), but also promoted calcium deposition and ALP of hBMMCs (P<0.05). Conclusion A-PRF improved the osteogenic differentiation of hBMMCs and the proliferation and migration, and 10% A-PRF achieved the most obvious promotion effect. The favorable cytobiological effects of A-PRF on hBMMCs might serve as the basis for A-PRF applications in bone regeneration.

Key words: Advanced-platelet rich fibrin, Human bone marrow mesenchymal cells, Osteogenic differentiation

Wang Jia, Zhao Ruimin, Gong Jiaming, Yu Zhanhai, Yin Lihua. Effect of A-PRF on the biological performance of human bone marrow mesenchymal cells[J]. Chinese Journal of Oral Implantology, 2021, 26(5): 282-288.

References

[1] Dawson JI, Oreffo RO.Bridging the regeneration gap: stem cells, biomaterials and clinical translation in bone tissue engineering[J]. Arch Biochem Biophys, 2008,473(2):124-131. DOI: 10.1016/j.abb.2008.03. 024.
[2] Haugen HJ, Lyngstadaas SP, Rossi F, et al. Bone grafts: which is the ideal biomaterial?[J]. J Clin Periodontol, 2019,46 Suppl 21:92-102. DOI: 10.1111/jcpe.13058.
[3] Marx RE, Carlson ER, Eichstaedt RM, et al.Platelet-rich plasma: Growth factor enhancement for bone grafts[J]. Oral Surg Oral Med Oral Pathol Oral Radiol Endod, 1998,85(6):638-646. DOI: 10.1016/s1079-2104(98) 90029-4.
[4] Graziani F, Ivanovski S, Cei S, et al.The in vitro effect of different PRP concentrations on osteoblasts and fibroblasts[J]. Clin Oral Implants Res, 2006,17(2):212-219. DOI: 10.1111/j.1600-0501.2005.01203.x.
[5] Kobayashi E, Flückiger L, Fujioka-Kobayashi M, et al.Comparative release of growth factors from PRP, PRF, and advanced-PRF[J]. Clin Oral Investig, 2016,20(9):2353-2360. DOI: 10.1007/s00784-016-1719-1.
[6] Masuki H, Okudera T, Watanebe T, et al.Growth factor and pro-inflammatory cytokine contents in platelet-rich plasma (PRP), plasma rich in growth factors (PRGF), advanced platelet-rich fibrin (A-PRF), and concentrated growth factors (CGF)[J]. Int J Implant Dent, 2016,2(1):19. DOI: 10.1186/s40729-016-0052-4.
[7] Su CY, Kuo YP, Tseng YH, et al.In vitro release of growth factors from platelet-rich fibrin (PRF): a proposal to optimize the clinical applications of PRF[J]. Oral Surg Oral Med Oral Pathol Oral Radiol Endod, 2009,108(1):56-61. DOI: 10.1016/j.tripleo.2009.02.004.
[8] 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. DOI: 10.1563/aaid-joi-D -14-00138.
[9] Lei L, Yu Y, Han J, et al.Quantification of growth factors in advanced platelet-rich fibrin and concentrated growth factors and their clinical efficacy as adjunctive to the GTR procedure in periodontal intrabony defects[J]. J Periodontol, 2020,91(4):462-472. DOI: 10.1002/JPER.19- 0290.
[10] Miron RJ, Zucchelli G, Pikos MA, et al.Use of platelet-rich fibrin in regenerative dentistry: a systematic review[J]. Clin Oral Investig, 2017,21(6):1913-1927. DOI: 10.1007/s00784-017-2133-z.
[11] Chang YR, Liu C, Yin LH.Research progress on platelet-rich fibrin derivatives[J]. Hua Xi Kou Qiang Yi Xue Za Zhi, 2019,37(6):660-665. DOI: 10.7518/hxkq.2019.06.016.
[12] Verma UP, Yadav RK, Dixit M, et al.Platelet-rich fibrin: a paradigm in periodontal therapy - a systematic review[J]. J Int Soc Prev Community Dent, 2017,7(5):227-233. DOI: 10.4103/jispcd.JISPCD_429_16.
[13] 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. DOI: 10.1563/aaid-joi-D-14-00138.
[14] Li X, Yang H, Zhang Z, et al.Platelet-rich fibrin exudate promotes the proliferation and osteogenic differentiation of human periodontal ligament cells in vitro[J]. Mol Med Rep, 2018,18(5):4477-4485. DOI: 10.3892/mmr.2018.9472.
[15] Verboket R, Herrera-Vizcaíno C, Thorwart K, et al.Influence of concentration and preparation of platelet rich fibrin on human bone marrow mononuclear cells (in vitro)[J]. Platelets, 2019,30(7):861-870. DOI: 10.1080/09537104.2018.1530346.
[16] Duan J, Kuang W, Tan J, et al.Differential effects of platelet rich plasma and washed platelets on the proliferation of mouse MSC cells[J]. Mol Biol Rep, 2011,38(4):2485-2490. DOI: 10.1007/s11033-010- 0385-7.
[17] Fujioka-Kobayashi M, Miron RJ, Hernandez M, et al.Optimized platelet-rich fibrin with the low-speed concept: growth factor release, biocompatibility, and Cellular Response[J]. J Periodontol, 2017,88(1): 112-121. DOI: 10.1902/jop.2016.160443.
[18] Cabaro S, D'Esposito V, Gasparro R, et al. White cell and platelet content affects the release of bioactive factors in different blood-derived scaffolds[J]. Platelets, 2018,29(5):463-467. DOI: 10.1080/09537104. 2017.1319046.
[19] Marques RE, Guabiraba R, Russo RC, et al.Targeting CCL5 in inflammation[J]. Expert Opin Ther Targets, 2013,17(12):1439-1460. DOI: 10.1517/14728222.2013.837886.
[20] Kim SJ, Kim SY, Kwon CH, et al.Differential effect of FGF and PDGF on cell proliferation and migration in osteoblastic cells[J]. Growth Factors, 2007,25(2):77-86. DOI: 10.1080/08977190701398977.
[21] Mehrotra M, Krane SM, Walters K, et al.Differential regulation of platelet-derived growth factor stimulated migration and proliferation in osteoblastic cells[J]. J Cell Biochem, 2004,93(4):741-752. DOI: 10.1002/jcb.20138.
[22] Li F, Yang Y, Zhu P, et al.Echinacoside promotes bone regeneration by increasing OPG/RANKL ratio in MC3T3-E1 cells[J]. Fitoterapia, 2012, 83(8):1443-1450. DOI: 10.1016/j.fitote.2012.08.008.
[23] Dohan Ehrenfest DM, Doglioli P, de Peppo GM, et al. Choukroun's platelet-rich fibrin (PRF) stimulates in vitro proliferation and differentiation of human oral bone mesenchymal stem cell in a dose- dependent way[J]. Arch Oral Biol, 2010,55(3):185-194. DOI: 10.1016/ j.archoralbio.2010.01.004.
[24] Nguyen N, Lee SB, Lee YS, et al.Neuroprotection by NGF and BDNF against neurotoxin-exerted apoptotic death in neural stem cells are mediated through Trk receptors, activating PI3-kinase and MAPK pathways[J]. Neurochem Res, 2009,34(5):942-951. DOI: 10.1007/s11064- 008-9848-9.
[25] Chao MV, Rajagopal R, Lee FS.Neurotrophin signalling in health and disease[J]. Clin Sci (Lond), 2006,110(2):167-173. DOI: 10.1042/ CS20050163.
[26] Chen WH, Mao CQ, Zhuo LL, et al.Beta-nerve growth factor promotes neurogenesis and angiogenesis during the repair of bone defects[J]. Neural Regen Res, 2015,10(7):1159-1165. DOI: 10.4103/1673-5374.160114.
[27] Zhang R, Liang Y, Wei S.The expressions of NGF and VEGF in the fracture tissues are closely associated with accelerated clavicle fracture healing in patients with traumatic brain injury[J]. Ther Clin Risk Manag, 2018,14:2315-2322. DOI: 10.2147/TCRM.S182325.
[28] 崔国胜, 曾剑玉, 汤晓飞,等. 神经生长因子对大鼠骨髓基质细胞成骨向分化的影响[J].口腔颌面修复学杂志. 2012, 13(05): 261-265.DOI:10.3969/ j.issn.1009-3761.2012.05.002
[29] 卢萌, 卓丽莉, 茅传青, 等. 外源性β-NGF对骨缺损区骨形成和改建的调节作用[J].中国口腔颌面外科杂志,2017,15(03):193-197.
[30] Hume DA.The mononuclear phagocyte system[J]. Curr Opin Immunol, 2006,18(1):49-53. DOI: 10.1016/j.coi.2005.11.008.
[31] Kong YY, Yoshida H, Sarosi I, et al.OPGL is a key regulator of osteoclastogenesis, lymphocyte development and lymph-node organogenesis[J]. Nature, 1999,397(6717):315-323. DOI: 10.1038/16852.