ABSTRACT
STATEMENT OF PROBLEM: Platelet-rich plasma(PRP) is well known to be very effective method to stimulate and accelerate the healing of bone and soft tissue. However, there are few reports which deal with the mechanisms of the PRP on the activation of the osteoblasts. PURPOSE: This study was aimed to investigate the effect of growth factors in PRP on the activity of osteoblasts. MATERIAL AND METHOD: To evaluate the effect on human, human osteoblast cell line was cultured. PRP was extracted from the blood of a healthy volunteer. Using the recombinant growth factors of PDGF, TGF-beta, IGF-1, bFGF which are mainly found at bone matrix and their neutralizing antibody, the effect of PRP on the attachment and proliferation of osteoblasts was evaluated. To evaluate the autocrine and paracrine effects, conditioned media(CM) of PRP was made and compared with PRP. By the western blot analysis, the expression of growth factors in PRP, CM was examined. Cell morphology was compared by the light microscope. RESULTS: 1) The effects of CM on osteoblast were similar to the effects of PRP. 2) PRP, CM, recombinant TGF-beta, bFGF, IGF-1 showed significantly higher cellular attachment than control(P<0.05) in the cell attachment assay. In the cell proliferation assay, PRP, CM, recombinant TGF-beta, IGF-1, bFGF, PDGF increased significantly cell proliferation(P<0.01). Among the recombinant growth factors, IGF-1 showed the highest cellular attachment and proliferation. 3) In the western blot assay, bFGF, IGF-1, PDGF were equally expressed in PRP and CM. 4) The attachment of osteoblast cell decreased significantly after the addition of neutralizing antibody against TGF-beta, IGF-1(P<0.05). In the cell proliferation assay, the addition of neutralizing antibody against TGF-beta, bFGF, PDGF, IGF-1 decreased significantly the cellular proliferation(P<0.05). The amount of decreasing in the cell attachment and proliferation is the highest in ab-IGF-1. 5) The cells in control group were flattened and elongated with a few cellular processes in the a light microscope. But, the cells appeared as spherical, plump cells with well developed cellular processes in experimental groups. The cells in PRP and CM had more prominent developed features than recombinant growth factor groups. CONCLUSIONS: These findings imply that PRP maximize the cellular activity in early healing period using the synergistic effect, autocrine, paracrine effects of growth factors and increase the rate and degree of bone formation.