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


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.


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.
Dohan Ehrenfest
David M
DM
Department of Biomaterials, Institute for Clinical Sciences, The Sahlgrenska Academy at University of Gothenburg, Sweden. LoB5@mac.com
Doglioli
Pierre
P
de Peppo
Giuseppe M
GM
Del Corso
Marco
M
Charrier
Jean-Baptiste
JB
D000469:Alkaline Phosphatase / Q000032:analysis; D001672:Biocompatible Materials / Q000494:pharmacology*; D001792:Blood Platelets / Q000502:physiology*; D002113:Calcification, Physiologic / Q000502:physiology; D002452:Cell Count; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D018920:Coculture Techniques; D004396:Coloring Agents; D003601:Cytotoxicity Tests, Immunologic; D004305:Dose-Response Relationship, Drug; D005337:Fibrin / Q000494:pharmacology*; D006801:Humans; D007962:Leukocytes / Q000502:physiology; D008297:Male; D008437:Maxilla / Q000166:cytology*; D059630:Mesenchymal Stem Cells / Q000187:drug effects*; D008855:Microscopy, Electron, Scanning; D008875:Middle Aged; D010006:Osteoblasts / Q000187:drug effects; D010027:Osteotomy / Q000379:methods; D013053:Spectrophotometry; D013778:Tetrazolium Salts; D013844:Thiazoles; D013997:Time Factors; D020858:Tissue and Organ Harvesting / Q000379:methods; D014343:Trypan Blue
D001672:Biocompatible Materials; D004396:Coloring Agents; D013778:Tetrazolium Salts; D013844:Thiazoles; D005337:Fibrin; D000469:Alkaline Phosphatase; C022616:thiazolyl blue; D014343:Trypan Blue