RSS-Feed abonnieren

DOI: 10.1055/s-0045-1812112
Effect of Centrifugation Methods on the Regenerative Potential of Concentrated Platelet-Rich Fibrin: An In Vitro Study
Autoren
Abstract
Objective
A new type of platelet concentrate used in regenerative periodontal therapy, called concentrated platelet-rich fibrin (C-PRF), contains more growth factors than previous generations of platelet concentrate. Compared to fixed-angle centrifugation, a recently discovered technique called horizontal centrifugation can produce C-PRF with a more evenly distributed quantity of platelets and improved biological properties. This study aims to determine the effect of horizontal and fixed-angle centrifugation methods on the release of transforming growth factor β1 (TGF-β1) and platelet-derived growth factor AB (PDGF-AB), as well as on the migration and proliferation of osteoblast cells.
Materials and Methods
This study used two test groups: a horizontal group and a fixed-angle group. The release of growth factors TGF-β1 and PDGF-AB was measured at five time points: the first, third, seventh, 10th, and 14th days of incubation, using the enzyme-linked immunosorbent assay. To determine the effect of C-PRF type on osteoblast cell migration, a scratch wound healing assay was conducted 24 hours after incubation. Osteoblast cell proliferation was counted using a 450-nm microplate reader to determine the number of cells in the well plate on the first, third, and fifth days, with the Cell Counting Kit-8. The data were examined using two-way ANOVA (analysis of variance) and post hoc LSD (least significant difference).
Results
The findings indicated a statistically significant increase in the release of TGF-β1 and PDGF-AB in the horizontal group compared to the fixed-angle group over a 14-day period. The horizontal group's TGF-β1 release increased by a factor of 1.5 compared to the fixed-angle group, peaking on day 7. Following a 24-hour incubation period and microscopic examination, the horizontal group exhibited greater osteoblast migratory activity than the control group and the fixed-angle centrifugation group. Horizontal centrifugation C-PRF had a significantly greater effect on osteoblast proliferation than fixed-angle centrifugation. The horizontal group's proliferation rate surpassed that of the fixed-angle group by up to 1.5 times on the third day of observation.
Conclusion
Compared to fixed-angle centrifugation, the horizontal method of preparing C-PRF has a greater effect on the release of growth factors and the behavior of osteoblasts.
Keywords
horizontal centrifugation - fixed-angle centrifugation - concentrated platelet-rich fibrin - TGF-β1 - PDGF-AB - proliferation - migrationInformed Consent
For this study, informed written consent was provided to conduct the outlined experiments prior to the blood collection from volunteers.
Publikationsverlauf
Artikel online veröffentlicht:
07. November 2025
© 2025. The Author(s). This is an open access article published by Thieme under the terms of the Creative Commons Attribution License, permitting unrestricted use, distribution, and reproduction so long as the original work is properly cited. (https://creativecommons.org/licenses/by/4.0/)
Thieme Medical and Scientific Publishers Pvt. Ltd.
A-12, 2nd Floor, Sector 2, Noida-201301 UP, India
-
References
- 1 Tattan M. Periodontics the Complete Summary: Surgical Regenerative Therapy. 1st ed.. Batavia, IL: Quintessence Publishing; 2021: 118-128
- 2 Miron RJ, Choukroun J, Wiley J. Platelet Rich Fibrin in Regenerative Dentistry: Biological Background and Clinical Indications. 1st ed.. Hoboken, NJ: Wiley; 2017: 4-5
- 3 Rodriguez JC, Bustillos MR. Periodontics the Complete Summary: Tissue Engineering. 1st ed.. Batavia, IL: Quintessence Publishing; 2021: 136-138
- 4 Miron RJ, Estrin NE, Ahmad P. et al. Thirty years of autologous platelet concentrates: from platelet-rich plasma to platelet-rich fibrin. J Periodontal Res 2025; 0: 1-28
- 5 Miron RJ, Fujioka-Kobayashi M, Hernandez M. et al. Injectable platelet rich fibrin (i-PRF): opportunities in regenerative dentistry?. Clin Oral Investig 2017; 21 (08) 2619-2627
- 6 Miron RJ, Chai J, Zhang P. et al. A novel method for harvesting concentrated platelet-rich fibrin (C-PRF) with a 10-fold increase in platelet and leukocyte yields. Clin Oral Investig 2020; 24 (08) 2819-2828
- 7 Sari R, Sukorini U, Susilowati H, Suryono S. Comparative study on interdental papillae regeneration: leukocyte platelet-rich fibrin by-product versus hyaluronic acid injections in modified open gingival embrasure model. Eur J Dent 2025; (e-pub ahead of print)
- 8 Miron RJ, Chai J, Zheng S, Feng M, Sculean A, Zhang Y. A novel method for evaluating and quantifying cell types in platelet rich fibrin and an introduction to horizontal centrifugation. J Biomed Mater Res A 2019; 107 (10) 2257-2271
- 9 Fujioka-Kobayashi M, Kono M, Katagiri H. et al. Histological comparison of platelet rich fibrin clots prepared by fixed-angle versus horizontal centrifugation. Platelets 2021; 32 (03) 413-419
- 10 Wang X, Zhang Y, Choukroun J, Ghanaati S, Miron RJ. Effects of an injectable platelet-rich fibrin on osteoblast behavior and bone tissue formation in comparison to platelet-rich plasma. Platelets 2018; 29 (01) 48-55
- 11 Fujioka-Kobayashi M, Katagiri H, Kono M. et al. Improved growth factor delivery and cellular activity using concentrated platelet-rich fibrin (C-PRF) when compared with traditional injectable (i-PRF) protocols. Clin Oral Investig 2020; 24 (12) 4373-4383
- 12 Caruana A, Savina D, Macedo JP, Soares SC. From platelet-rich plasma to advanced platelet-rich fibrin: biological achievements and clinical advances in modern surgery. Eur J Dent 2019; 13 (02) 280-286
- 13 Margono A, Bagio DA, Yulianto I, Dewi SU. Changes in migratory speed rate of human dental pulp stromal cells cultured in advanced platelet-rich fibrin. Eur J Dent 2023; 17 (01) 91-96
- 14 Staehlke S, Rebl H, Nebe B. Phenotypic stability of the human MG-63 osteoblastic cell line at different passages. Cell Biol Int 2019; 43 (01) 22-32
- 15 Liang CC, Park AY, Guan JL. In vitro scratch assay: a convenient and inexpensive method for analysis of cell migration in vitro. Nat Protoc 2007; 2 (02) 329-333
- 16 Fujioka-Kobayashi M, Schaller B, Mourão CFAB, Zhang Y, Sculean A, Miron RJ. Biological characterization of an injectable platelet-rich fibrin mixture consisting of autologous albumin gel and liquid platelet-rich fibrin (Alb-PRF). Platelets 2021; 32 (01) 74-81
- 17 Zwittnig K, Kirnbauer B, Jakse N. et al. Growth factor release within liquid and solid PRF. J Clin Med 2022; 11 (17) 5070-5070
- 18 Kargarpour Z, Nasirzade J, Panahipour L, Miron RJ, Gruber R. Relative centrifugal force (RCF; G-force) affects the distribution of TGF-β in PRF membranes produced using horizontal centrifugation. Int J Mol Sci 2020; 21 (20) 7629
- 19 Eppley BL, Woodell JE, Higgins J. Platelet quantification and growth factor analysis from platelet-rich plasma: implications for wound healing. Plast Reconstr Surg 2004; 114 (06) 1502-1508
- 20 Ferreira Sávio DS, Silva LMPD, Reis GGD. et al. Effects of platelet-rich fibrin produced by three centrifugation protocols on bone neoformation in defects created in rat calvaria. Platelets 2023; 34 (01) 2228417
- 21 Lourenço ES, Alves GG, de Lima Barbosa R. et al. Effects of rotor angle and time after centrifugation on the biological in vitro properties of platelet rich fibrin membranes. J Biomed Mater Res B Appl Biomater 2021; 109 (01) 60-68
- 22 Gheno E, Mourão CFAB, Mello-Machado RC. et al. In vivo evaluation of the biocompatibility and biodegradation of a new denatured plasma membrane combined with liquid PRF (Alb-PRF). Platelets 2021; 32 (04) 542-554
- 23 Al-Maawi S, Dohle E, Kretschmer W, Rutkowski J, Sader R, Ghanaati S. A standardized g-force allows the preparation of similar platelet-rich fibrin qualities regardless of rotor angle. Tissue Eng Part A 2022; 28 (7-8): 353-365
- 24 Mu Z, He Q, Xin L. et al. Effects of injectable platelet rich fibrin on bone remodeling in combination with DBBM in maxillary sinus elevation: a randomized preclinical study. Am J Transl Res 2020; 12 (11) 7312-7325
- 25 Gollapudi M, Bajaj P, Oza RR. Injectable platelet-rich fibrin - a revolution in periodontal regeneration. Cureus 2022; 14 (08) e28647
- 26 Fiedler J, Röderer G, Günther KP, Brenner RE. BMP-2, BMP-4, and PDGF-bb stimulate chemotactic migration of primary human mesenchymal progenitor cells. J Cell Biochem 2002; 87 (03) 305-312
- 27 Azizi A. Designing of artificial intelligence model-free controller based on output error to control wound healing process. Biosens J 2017; 6 (01) 1-19
- 28 Farshidfar N, Apaza Alccayhuaman KA, Estrin NE. et al. Advantages of horizontal centrifugation of platelet-rich fibrin in regenerative medicine and dentistry. Periodontol 2000 2025; 00: 1-52
