Choukroun's platelet-rich fibrin (PRF) stimulates in vitro proliferation and differentiation of human oral bone mesenchymal stem cell in a dose-dependent way

Dohan Ehrenfest, David M.; Doglioli, Pierre; de Peppo, Giuseppe M.; Del Corso, Marco; Charrier, Jean-Baptiste

doi:10.1016/j.archoralbio.2010.01.004

« Previous Next »Archives of Oral Biology
Volume 55, Issue 3 , Pages 185-194, March 2010

Choukroun's platelet-rich fibrin (PRF) stimulates in vitro proliferation and differentiation of human oral bone mesenchymal stem cell in a dose-dependent way

David M. Dohan Ehrenfest
- Department of Biomaterials, Institute for Clinical Sciences, The Sahlgrenska Academy at University of Gothenburg, Sweden
- The LoB5 Foundation for Research, Paris, France
- Corresponding author at: Department of Biomaterials, Institute for Clinical Sciences, The Sahlgrenska Academy at University of Gothenburg, Medicinaregatan 8B, 41390 Gothenburg, Sweden. Tel.: +33 6 64 72 95 40.
Pierre Doglioli
- Cell Culture Laboratory, Jules Ferry Institute, Cannes, France
Giuseppe M. de Peppo
- Department of Biomaterials, Institute for Clinical Sciences, The Sahlgrenska Academy at University of Gothenburg, Sweden
Marco Del Corso
- The LoB5 Foundation for Research, Paris, France
Jean-Baptiste Charrier
- The LoB5 Foundation for Research, Paris, France
- Department of ENT, Head and Neck Surgery, Paris Sud University, AP-HP Bicêtre Hospital, Le Kremlin Bicêtre, France

Accepted 15 January 2010.

Abstract

Background

Choukroun's platelet-rich fibrin (PRF) is an autologous leukocyte- and platelet-rich fibrin biomaterial. The purpose of this study was to analyse the in vitro effects of PRF on human bone mesenchymal stem cells (BMSC), harvested in the oral cavity after preimplant endosteal stimulation.

Materials and methods

BMSCs from primary cultures were cultivated with or without a PRF membrane originating from the same donor as for the cells, in proliferation or osteoblastic differentiation conditions. After 7 days, the PRF membranes were removed. A series of cultures were performed using 2 PRF membranes, in order to measure the dose-dependent effect. Cell counts, cytotoxicity tests, alkaline phosphatase (ALP) activity quantification, Von Kossa staining and mineralisation nodules counts were performed at 3, 7, 14, 21 and 28 days. A last independent series was carried on up to 14 days, for a morphological scanning electron microscope (SEM) observation.

Results

PRF generated a significant stimulation of the BMSC proliferation and differentiation throughout the experimental period. This effect was dose-dependent during the first weeks in normal conditions, and during the whole experimentation in differentiation conditions. The cultures without PRF in differentiation conditions did not rise above the degree of differentiation of the cultures in normal conditions with 1 or 2 PRF up to the 14th and 28th day, respectively. The SEM culture analysis at day 14 allowed to show the mineralisation nodules which were more numerous and more structured in the groups with PRF compared to the control groups.

Discussion and conclusions

This double contradictory proliferation/differentiation result may be due to the numerous components of PRF, particularly the presence of leukocytes: any culture with PRF is in fact a coculture with leukocytes. It could be the source of differential geographic regulation processes within the culture. The combination of oral BMSC and PRF might offer many potential clinical and biotechnological applications, and deserves new studies.

Keywords: Coculture, Growth factors, Leukocyte, Platelet concentrate, Mesenchymal stem cell, Platelet-rich fibrin (PRF), Platelet-rich plasma (PRP)