The in vitro effect of different PRP concentrations on osteoblasts and fibroblasts.
OBJECTIVE
The aim of this study was to assess the biological rationale for the use of platelet-rich plasma (PRP) by evaluating the effect of different concentrations of PRP on osteoblasts (OB) and fibroblasts (FB) function in vitro.


METHODS
PRP was obtained from volunteer donors using standard protocols. Primary human cultures of oral FBs and OBs were exposed to both activated and non-activated plasma as well as various concentrations of PRP (2.5 x, 3.5 x and max (4.2-5.5 x)). Cell proliferation was evaluated after 24 and 72 h using an MTT proliferation assay. Production of osteocalcin (OCN), osteoprotegerin (OPG) and transforming growth factor beta1 (TGF-beta1) was evaluated in OB after 24 and 72 h. Statistical analysis was performed using one-way ANOVA.


RESULTS
PRP-stimulated cell proliferation in both OBs and FBs. The effect of different PRP concentrations on cell proliferation was most notable at 72 h. The maximum effect was achieved with a concentration of 2.5 x, with higher concentrations resulting in a reduction of cell proliferation. Upregulation of OCN levels and downregulation of OPG levels were noted with increasing PRP concentrations at both 24 and 72 h. TGF-beta1 levels were stimulated by increasing concentrations of PRP, with the increased levels being maintained at 72 h.


CONCLUSIONS
PRP preparations exert a dose-specific effect on oral FBs and OBs. Optimal results were observed at a platelet concentration of 2.5 x, which was approximately half of the maximal concentrate that could be obtained. Increased concentrations resulted in a reduction in proliferation and a suboptimal effect on OB function. Hence, different PRP concentrations may have an impact on the results that can be obtained in vivo.
Department of Neurosciences, Section of Oral Surgery, University of Pisa, Pisa, Italy. f.graziani@eastman.ucl.ac.uk
Filippo
F
Graziani
Saso
S
Ivanovski
Silvia
S
Cei
Francesco
F
Ducci
Maurizio
M
Tonetti
Mario
M
Gabriele
D000328:Adult; D001792:Blood Platelets / Q000502:physiology*; D001842:Bone and Bones; D049109:Cell Proliferation; D002478:Cells, Cultured; D015536:Down-Regulation; D005260:Female; D005347:Fibroblasts / Q000502:physiology*; D005881:Gingiva; D006023:Glycoproteins / Q000032:analysis; D006801:Humans; D008875:Middle Aged; D010006:Osteoblasts / Q000502:physiology*; D015675:Osteocalcin / Q000032:analysis; D053244:Osteoprotegerin; D010949:Plasma; D018160:Receptors, Cytoplasmic and Nuclear / Q000032:analysis; D018124:Receptors, Tumor Necrosis Factor / Q000032:analysis; D013997:Time Factors; D016212:Transforming Growth Factor beta / Q000032:analysis; D053773:Transforming Growth Factor beta1; D015854:Up-Regulation
D006023:Glycoproteins; D053244:Osteoprotegerin; D018160:Receptors, Cytoplasmic and Nuclear; D018124:Receptors, Tumor Necrosis Factor; C508960:TGFB1 protein, human; C506030:TNFRSF11B protein, human; D016212:Transforming Growth Factor beta; D053773:Transforming Growth Factor beta1; D015675:Osteocalcin