The impact of storage protocols on the mechanical and structural properties of frozen platelet-rich fibrin membranes
DOI:
https://doi.org/10.2340/biid.v12.44903Keywords:
Elasticity, platelet-rich-fibrin, scanning electron microscopy, tensile strengthAbstract
Background & objectives: Platelet-rich fibrin (PRF) has shown promise in periodontal surgery and is found to promote bone regeneration. However, its limitations include low elasticity, short lifespan, and poor storage stability. This study aimed to overcome these limitations by developing frozen PRF and comparing the mechanical and structural properties of advanced-PRF (A-PRF) and two frozen PRF storage protocols.
Methodology: This in vitro study used blood samples from 14 healthy volunteers to prepare A-PRF, which was stored at −20°C for 1 day (Fz-PRF1) or 7 days (Fz-PRF7). The study evaluated mechanical properties (tensile strength and elasticity) using a universal testing machine, and structural properties (Blood Elements Adhesion Index [BEAI] and fibrin density) using scanning electron microscopy (SEM) analysis. Results were analysed by Chi-square, independent t-tests, and Tukey post-hoc tests.
Results: Comparing three PRF membranes, Fz-PRF7 showed significantly higher tensile strength than A-PRF and Fz-PRF1, and higher modulus of elasticity than A-PRF. However, SEM analysis revealed no significant differences in BEAI and fibrin density scores across groups.
Conclusion: Fz-PRF7 showed improved mechanical properties, specifically tensile strength and elasticity, compared to A-PRF, overcoming some limitations of conventional PRF.
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Liang Y, Luan X, Liu X. Recent advances in periodontal regeneration: a biomaterial perspective. Bioact Mater. 2020;5(2):297–308. https://doi.org/10.1016/j.bioactmat.2020.02.012 DOI: https://doi.org/10.1016/j.bioactmat.2020.02.012
Daghrery A, Bottino MC. Advanced biomaterials for periodontal tissue regeneration. Genesis. 2022;60:e23501. https://doi.org/10.1002/dvg.23501 DOI: https://doi.org/10.1002/dvg.23501
Quirynen M, Siawasch S, Temmerman A, Cortellini S, Dhondt R, Teughels W, et al. Do autologous platelet concentrates (APCs) have a role in intra-oral bone regeneration? A critical review of clinical guidelines on decision-making process. Perio-dontol 2000. 2023;93(1):254–69. https://doi.org/10.1111/prd.12526 DOI: https://doi.org/10.1111/prd.12526
Fujioka-Kobayashi M, Miron RJ, Hernandez M, Kandalam U, Zhang Y, Choukroun J. Optimized platelet-rich fibrin with the low-speed concept: growth factor release, biocompatibility, and cellular response. J Periodontol. 2017;88(1):112–21. https://doi.org/10.1902/jop.2016.160443 DOI: https://doi.org/10.1902/jop.2016.160443
Kob Kobayashi E, Flückiger L, Fujioka-Kobayashi M, Sawada K, Sculean A, Schaller B, et al. Comparative release of growth factors from PRP, PRF, and advanced-PRF. Clin Oral Investig. 2016;20(9):2353–60. https://doi.org/10.1007/s00784-016-1719-1 DOI: https://doi.org/10.1007/s00784-016-1719-1
Choukroun J, Diss A, Simonpieri A, Girard MO, Schoeffler C, Dohan SL, et al. Platelet-rich fibrin (PRF): a second-generation platelet concentrate. Part IV: clinical effects on tissue healing. Oral Surg Oral Med Oral Pathol Oral Radiol Endodontol. 2006;101:e56–60. DOI: https://doi.org/10.1016/j.tripleo.2005.07.011
Zwittnig K, Kirnbauer B, Jakse N, Schlenke P, Mischak I, Ghanaati S, et al. Growth factor release within liquid and solid PRF. J Clin Med. 2022;11(17):5070. https://doi.org/10.3390/jcm11175070 DOI: https://doi.org/10.3390/jcm11175070
Lambrichts I, Wolfs E, Bronckaers A, Gervois P, Vangansewinkel T. The effect of leukocyte- and platelet-rich fibrin on central and peripheral nervous system neurons-implications for biomaterial applicability. Int J Mol Sci. 2023;24(18):14314. https://doi.org/10.3390/ijms241814314 DOI: https://doi.org/10.3390/ijms241814314
Dohan DM, Choukroun J, Diss A, Dohan SL, Dohan AJ, Mouhyi J, et al. Platelet-rich fibrin (PRF): a second generation platelet concentrate. Part II: platelet-related biologic features. Oral Surg Oral Med Oral Pathol Oral Radiol Endodontol. 2006;101:e45–50. DOI: https://doi.org/10.1016/j.tripleo.2005.07.009
Miron RJ, Choukroun J. Platelet rich fibrin in regenerative dentistry. Hoboken, NJ: John Wiley & Sons Ltd.; 2017. DOI: https://doi.org/10.1002/9781119406792.ch4
Miron RJ, Fujioka-Kobayashi M, Sculean A, Zhang Y. Optimization of platelet-rich fibrin. Periodontol 2000. 2024;94:79–91. https://doi.org/10.1111/prd.12521 DOI: https://doi.org/10.1111/prd.12521
Miron RJ, Pikos MA, Estrin NE, Kobayashi-Fujioka M, Espinoza AR, Basma H, et al. Extended platelet-rich fibrin. Periodontol 2000. 2024;94(1):114–30. https://doi.org/10.1111/prd.12537 DOI: https://doi.org/10.1111/prd.12537
Ngah NA, Ratnayake J, Cooper PR, Dias GJ, Tong DC, Mohd Noor SNF, et al. Potential of lyophilized platelet concentrates for craniofacial tissue regenerative therapies. Molecules. 2021;26:517. https://doi.org/10.3390/molecules26030517 DOI: https://doi.org/10.3390/molecules26030517
Xu F, Zou D, Dai T, Xu H, An R, Liu Y, et al. Effects of incorporation of granule-lyophilised platelet-rich fibrin into polyvinyl alcohol hydrogel on wound healing. Sci Rep. 2018;8:14042. https://doi.org/10.1038/s41598-018-32208-5 DOI: https://doi.org/10.1038/s41598-018-32208-5
Ngah NA, Dias GJ, Tong DC, Mohd Noor SNF, Ratnayake J, Cooper PR, et al. Lyophilised platelet-rich fibrin: physical and bio-logical characterisation. Molecules. 2021;26(23):7131. https://doi.org/10.3390/molecules26237131. DOI: https://doi.org/10.3390/molecules26237131
Isobe K, Suzuki M, Watanabe T, Kitamura Y, Suzuki T, Kawabata H, et al. Platelet-rich fibrin prepared from stored whole-blood samples. Int J Implant Dent. 2017;3(1):6. https://doi.org/10.1186/s40729-017-0068-4 DOI: https://doi.org/10.1186/s40729-017-0068-4
Ockerman A, Braem A, EzEldeen M, Castro A, Coucke B, Politis C, et al. Mechanical and structural properties of leukocyte- and platelet-rich fibrin membranes: an in vitro study on the impact of anticoagulant therapy. J Periodontal Res. 2020;55(5):686–93. https://doi.org/10.1111/jre.12755 DOI: https://doi.org/10.1111/jre.12755
Dias FJ, Venegas C, Borie E, Arias A, Watanabe IS, Fuentes R. A new insight of Platelet-Rich Fibrin clots morphology and their elemental composition. Tissue Cell. 2020;65:101362. https://doi.org/10.1016/j.tice.2020.101362 DOI: https://doi.org/10.1016/j.tice.2020.101362
Nascimento G, Leite AA, Leite ERM, Sampaio JEC, Leite FRM. Blood clot stabilization on root dentin conditioned by the com-bination of tetracycline and EDTA. Braz J Oral Sci. 2014;13(2):83–8. https://doi.org/10.1590/1677-3225v13n2a01 DOI: https://doi.org/10.1590/1677-3225v13n2a01
Dantas AA, Fontanari LA, Ishi Ede P, Leite FRM, Zandim-Barcelos DL, Rached RSGA, et al. Blood cells attachment after root conditioning and PRP application: an in vitro study. J Contemp Dent Pract. 2012;13:332–8. https://doi.org/10.5005/jp-journals-10024-1147 DOI: https://doi.org/10.5005/jp-journals-10024-1147
Khorshidi H, Raoofi S, Bagheri R, Banihashemi H. Comparison of the mechanical properties of early leukocyte‐and platelet‐rich fibrin versus PRGF/endoret membranes. Int J Dent. 2016:2016:1849207. https://doi.org/10.1155/2016/1849207 DOI: https://doi.org/10.1155/2016/1849207
Anitua E, Sanchez M, Orive G, Andia I. The potential impact of the preparation rich in growth factors (PRGF) in different medical fields, Biomaterials. 2007;28(31):4551–60. https://doi.org/10.1016/j.biomaterials.2007.06.037 DOI: https://doi.org/10.1016/j.biomaterials.2007.06.037
Chhaya D, Vaidya N, Patel V, Chudasama K, Doshi SH, Kumar P. Evaluation and comparison of mechanical properties of platelet-rich fibrin membrane, fish collagen membrane, bovine collagen membrane and chorionic membrane – an SEM study. Indian J Dent Res. 2022;33(4):425–9. https://doi.org/10.4103/ijdr.ijdr_556_22 DOI: https://doi.org/10.4103/ijdr.ijdr_556_22
Petronilho VG, de Fátima Balderrama Í, de Oliveira LA, Queiroz PM, Zubek MG, Gottardo VD. Evaluation of mechanical proper-ties of platelet-rich fibrin membrane for implant surgery: an analysis in vitro. J Indian Soc Periodontol. 2022;26(1):19–23. https://doi.org/10.4103/jisp.jisp_782_20 DOI: https://doi.org/10.4103/jisp.jisp_782_20
Shi L, Li R, Wei S, Zhou M, Li L, Lin F, et al. Effects of a protective agent on freeze-dried platelet-rich plasma. Blood Coagul Fibrinolysis. 2019;30(2):58–65. https://doi.org/10.1097/MBC.0000000000000796 DOI: https://doi.org/10.1097/MBC.0000000000000796
Warin R, Vongchan P, Suriyasathaporn W, Boripun R, Suriyasathaporn W. In vitro assessment of lyophilized advanced plate-let-rich fibrin from dogs in promotion of growth factor release and wound healing. Vet Sci. 2022;9(10):566. https://doi.org/10.3390/vetsci9100566 DOI: https://doi.org/10.3390/vetsci9100566
Kardos D, Hornyák I, Simon M, Hinsenkamp A, Marschall B, Várdai R, et al. Biological and mechanical properties of platelet-rich fibrin membranes after thermal manipulation and preparation in a single-syringe closed system. Int J Mol Sci. 2018;19(11):3433. https://doi.org/10.3390/ijms19113433 DOI: https://doi.org/10.3390/ijms19113433
Uchiyama R, Omura H, Maehara M, Toyoda E, Tamaki M, Ogawa M. Effect of freeze-thawing treatment on platelet-rich plasma purified with different kits. Int J Mol Sci. 2024;25(18):9981. https://doi.org/10.3390/ijms25189981 DOI: https://doi.org/10.3390/ijms25189981
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Copyright (c) 2025 Sruthima N.V.S. Gottumukkala, Lois Sowmya Abhinandhitha Rayapati, Gautami S. Penmetsa, Konathala S.V. Ramesh, Mohan Kumar Pasupuleti, Anil Kumar Kanakamedala, Shanmukh Bandaru
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