In vitro effects of Choukroun's PRF (platelet-rich fibrin) on human gingival fibroblasts, dermal prekeratinocytes, preadipocytes, and maxillofacial osteoblasts in primary cultures.
OBJECTIVE
The objective of this study was to analyze the effects of Choukroun's PRF (platelet-rich fibrin), a leucocyte and platelet concentrate clinically usable as fibrin membrane or clot, on human primary cultures of gingival fibroblasts, dermal prekeratinocytes, preadipocytes, and maxillofacial osteoblasts.


METHODS
For the proliferation study, these cells were cultivated with or without a PRF membrane originating from the same donor as for the cells. For osteoblasts and fibroblasts, dose-dependent effect was assessed (using 2 membranes). Cell counts and cytotoxicity tests were performed at 3, 7, 14, and 21 days, and even 28 days for osteoblasts. More osteoblast cultures were prepared in differentiation conditions according to 3 modalities (without PRF, with PRF, with PRF the first day and differentiation medium applied only after the first week of culture). Osteoblast differentiation was analyzed using Von Kossa staining and alkaline phosphatase, DNA and total cell proteins dosage.


RESULTS
PRF induced a significant and continuous stimulation of proliferation in all cell types. It was dose dependent during all the experiment with osteoblasts, but only on day 14 with fibroblasts. Moreover, PRF induced a strong differentiation in the osteoblasts, whatever the culture conditions. The analysis of osteoblast cultures in differentiation conditions with PRF, using light and scanning electron microscopy, revealed a starting mineralization process in the PRF membrane itself after 14 days. Moreover, PRF leucocytes seemed to proliferate and interact with osteoblasts.


CONCLUSIONS
Cultures with PRF are always cocultures with leucocytes. These "chaperone leucocytes" could be the source of differential geographic regulation within the culture and explain the double contradictory effect proliferation/differentiation observed on osteoblasts.
Dohan Ehrenfest
David M
DM
Researcher, Department of Biomaterials, Institute for Clinical Sciences, The Sahlgrenska Academy at University of Gothenburg, Gothenburg, Sweden. LoB5@mac.com
Diss
Antoine
A
Odin
Guillaume
G
Doglioli
Pierre
P
Hippolyte
Marie-Pascale
MP
Charrier
Jean-Baptiste
JB
D017667:Adipocytes / Q000187:drug effects*; D000469:Alkaline Phosphatase / Q000032:analysis; D015415:Biomarkers / Q000032:analysis; D001792:Blood Platelets / Q000502:physiology*; D002113:Calcification, Physiologic / Q000187:drug effects; 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; D004305:Dose-Response Relationship, Drug; D005337:Fibrin / Q000008:administration & dosage / Q000494:pharmacology* / Q000648:ultrastructure; D005347:Fibroblasts / Q000187:drug effects*; D005881:Gingiva / Q000166:cytology* / Q000187:drug effects; D006801:Humans; D015603:Keratinocytes / Q000187:drug effects*; D007962:Leukocytes / Q000502:physiology*; D008297:Male; D008334:Mandible / Q000166:cytology; D008855:Microscopy, Electron, Scanning; D008875:Middle Aged; D010006:Osteoblasts / Q000187:drug effects*; D012867:Skin / Q000166:cytology* / Q000187:drug effects; D013778:Tetrazolium Salts; D013844:Thiazoles; D013997:Time Factors
D015415:Biomarkers; D004396:Coloring Agents; D013778:Tetrazolium Salts; D013844:Thiazoles; D005337:Fibrin; D000469:Alkaline Phosphatase; C022616:thiazolyl blue