The effects of platelet-rich plasma on cell cultures of oral cells related with regeneration

• Main Authors: Chia-Wen Hsu, 許嘉文
• Other Authors: Kuo Yuan
• Format: Others
• Language: zh-TW
• Published: 2006
• Online Access: http://ndltd.ncl.edu.tw/handle/62144077682415171908

碩士 === 國立成功大學 === 口腔醫學研究所 === 94 === Platelets are rapidly activated by blood vessel injury and release products from the alpha granules containing growth factors, thrombospondin, fibrinogen, etc. The growth factors include platelet-derived growth factor (PDGF) and transforming growth factor-β (TGF-β), which can enhance wound healing. Platelet-rich plasma (PRP) is derived from autologous blood by sequestering and concentrating the platelets via centrifugation. PRP gel is an autologous modification of fibrin glue that has been described and used in various applications, but its clinical results remained controversial. We wanted to know whether a negative regulator exists in PRP, because the major function of thrombospondin-1 (TSP-1) is inhibiting angiogenesis. The aim of this study was to evaluate the regulators on PRP effects and its biological function in vitro. Twenty volunteers participated in this study and donated their venous blood. Using centrifugation, platelet-poor plasma (PPP) and PRP containing 100-120 × 104/μl platelets were obtained. We added different concentrations of fresh PRP and PPP to cell cultures related to regeneration: periodontal ligament (PDL) fibroblast, osteoblast, and endothelial cell cultures. We measured the rate of cell proliferation using an alamar blue assay for 6 days. Subsequently, after protein concentrations had been determined, the band density of TSP-1 was examined using Western blotting. The quantitation of TSP-1 was estimated using an ELISA assay. Finally, we incubated the cell cultures for 2 days in serial concentrations of TSP-1 that corresponded to 30% PRP gel. 3 We found that, after 6 days of incubation, the cells increased in a low concentration of PRP (< 5%) and decreased in a high concentration (15-30%). Western blotting showed that TSP-1 expression was higher in PRP than in PPP. An ELISA assay showed that the quantities of TSP-1 expressed were 183.3 ± 21.6 μg/ml in PRP and 9.7 ± 1.6 μg/ml in the supernatant of 30% PRP gel. The cell culture in serial concentrations of TSP-1 dose-dependently decreased. Within the limits of the present study, it can be concluded that the number of cells significantly decreased when treated with a high concentration of PRP and that PRP is abundant in TSP-1, which may play an important role in apoptosis.