Small Extracellular Vesicles of Platelet-rich Plasma Promoted Bone Tissue Repair

doi:10.13701/j.cnki.kqyxyj.2025.05.012

Abstract

Abstract: Objective: To investigate the impact of platelet-rich plasma derived small extracellular vesicles (PRP-sEV) on the expression of bone markers in osteoblasts, as well as to observe the bone repair effect of PRP-sEV in a mouse calvarial defect model. Methods: The small extracellular vesicles were extracted from mouse platelet-rich plasma. These PRP-sEV were then analyzed for size, structure, and marker proteins using nanoparticle tracking analysis, transmission electron microscopy, and western blot. PKH67 was used to assess the uptake of these vesicles by MC3T3-E1 cells and used western blot to examine the expression of osteogenic marker proteins. A mouse cranial defect model was established and evaluated the bone repair effects of PRP-sEV through hematoxylin-eosin (HE) staining and Masson staining. Immunohistochemistry was performed to assess the expression of bone morphogenetic protein 2 (BMP-2) and osteopontin (OPN) in PRP-sEV in the cranial defect model. Results: The size of PRP-sEV was around 144 nm. Transmission electron microscopy revealed that these vesicles had a typical lipid bilayer structure. Western blot results indicated that the positive marker proteins CD63, CD81, and ALG-2-interacting protein X (ALIX) were present in the vesicles, while the surface marker protein CD40 was absent. PKH67 testing demonstrated that MC3T3-E1 cells effectively incepted the PRP-sEV. Western blot results showed that these vesicles enhanced the expression of bone formation markers BMP-2 and OPN (P<0.05). HE staining and Masson staining results indicated that the PRP-sEV promoted bone defect repair. Immunohistochemistry results demonstrated that the expression of BMP-2 and OPN in the cranial bone defect model treated with the vesicles was higher than that of control group (P<0.05). Conclusion: The extracellular vesicles in platelet-rich plasma could promote bone tissue repair.

Key words: cranial bone defect model, platelet-rich plasma, extracellular vesicles, bone repair

GAO Yang, GAO Linbo, SHI Chun, WU Dalei. Small Extracellular Vesicles of Platelet-rich Plasma Promoted Bone Tissue Repair[J]. Journal of Oral Science Research, 2025, 41(5): 426-431.

References

[1] 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.
[2] Simonpieri A, Del Corso M, Vervelle A, et al. Current knowledge and perspectives for the use of platelet-rich plasma (PRP) and platelet-rich fibrin (PRF) in oral and maxillofacial surgery part 2: Bone graft, implant and reconstructive surgery [J]. Curr Pharm Biotechnol, 2012, 13(7): 1231-1256.
[3] Gholami L, Nooshabadi VT, Shahabi S, et al. Extracellular vesicles in bone and periodontal regeneration: current and potential therapeutic applications [J]. Cell Biosci, 2021, 11(1): 16.
[4] Jin Y, Long D, Li J, et al. Extracellular vesicles in bone and tooth: A state-of-art paradigm in skeletal regeneration [J]. J Cell Physiol, 2019, 234(9): 14838-14851.
[5] Liu M, Sun Y, Zhang Q. Emerging role of extracellular vesicles in bone remodeling [J]. J Dent Res, 2018, 97(8): 859-868.
[6] Tao SC, Guo SC. Extracellular vesicles in bone: "dogrobbers" in the "eternal battle field" [J]. Cell Commun Signal, 2019, 17(1): 6.
[7] Sun J, Hu Y, Fu Y, et al. Emerging roles of platelet concentrates and platelet-derived extracellular vesicles in regenerative periodontology and implant dentistry [J]. APL Bioeng, 2022, 6(3): 031503.
[8] Dong M, Shi C, Yu X, et al. Milk-derived small extracellular vesicles: nanomaterials to promote bone formation [J]. J Nanobiotechnology, 2022, 20(1): 370.
[9] Bennell KL, Hunter DJ, Paterson KL. Platelet-rich plasma for the management of hip and knee osteoarthritis [J]. Curr Rheumatol Rep, 2017, 19(5): 24.
[10] Oryan A, Alidadi S, Moshiri A. Platelet-rich plasma for bone healing and regeneration [J]. Expert Opin Biol Ther, 2016, 16(2): 213-232.
[11] Tang R, Wang S, Yang J, et al. Application of platelet-rich plasma in traumatic bone infections [J]. Expert Rev Anti Infect Ther, 2021, 19(7): 867-875.
[12] Nallakumarasamy A, Jeyaraman M, Maffulli N, et al. Mesenchymal stromal cell-derived extracellular vesicles in wound healing [J]. Life (Basel), 2022, 12(11): 1733.
[13] Shimizu Y, Ntege EH, Sunami H. Current regenerative medicine-based approaches for skin regeneration: A review of literature and a report on clinical applications in Japan [J]. Regen Ther, 2022, 21: 73-80.
[14] Jansen EE, Braun A, Jansen P, et al. Platelet-therapeutics to improve tissue regeneration and wound healing-physiological background and methods of preparation [J]. Biomedicines, 2021, 9(8):869.
[15] Rodríguez-Eguren A, Gómez-álvarez M, Francés-Herrero E, et al. Human umbilical cord-based therapeutics: Stem cells and blood derivatives for female reproductive medicine [J]. Int J Mol Sci, 2022, 23(24): 15942.
[16] Becker RC, Sexton T, Smyth SS. Translational implications of platelets as vascular first responders [J]. Circ Res, 2018, 122(3): 506-522.
[17] Wu J, Piao Y, Liu Q, et al. Platelet-rich plasma-derived extracellular vesicles: A superior alternative in regenerative medicine? [J]. Cell Prolif, 2021, 54(12): e13123.
[18] Spakova T, Janockova J, Rosocha J. Characterization and therapeutic use of extracellular vesicles derived from platelets [J]. Int J Mol Sci, 2021, 22(18): 9701.