Biomaterial Database

Freeze-Dried Platelet-Rich Plasma Induces Osteoblast Proliferation via Platelet-Derived Growth Factor Receptor-Mediated Signal Transduction. 2020

Hideyuki Kinoshita, and Sumihisa Orita, and Kazuhide Inage, and Kazuki Fujimoto, and Yasuhiro Shiga, and Koki Abe, and Masahiro Inoue, and Masaki Norimoto, and Tomotaka Umimura, and Takeshi Ishii, and Tsukasa Yonemoto, and Hiroto Kamoda, and Toshinori Tsukanishi, and Masahiko Suzuki, and Naoya Hirosawa, and Tsutomu Akazawa, and Seiji Ohtori

Department of Orthopaedic Surgery, Graduate School of Medicine, Chiba University, Chiba, Japan.

METHODS Controlled laboratory study. OBJECTIVE This study aimed to evaluate the in vitro pharmacological activity of growth factors (GFs) in freeze-dried platelet-rich plasma (FD-PRP) after storage for 4 weeks. BACKGROUND Freshly prepared PRP is a rich source of many GFs. We reported that FD-PRP stored for 8 weeks accelerated bone union in a rat posterolateral fusion model equally well as fresh-PRP. However, the pharmacological activity of FD-PRP after longterm storage has not been shown in vitro. METHODS Immediately after preparation, as well as 4 weeks after freeze-dried storage, the platelet count was measured. Human osteoblasts were treated with fresh-PRP and FD-PRP, respectively. Western blotting was used to assess the phosphorylation of the platelet-derived growth factor (PDGF) receptor (PDGFR) and its downstream target, extracellular signal-regulated kinase (ERK). The proliferation rates of osteoblasts were investigated by immunocytochemistry and MTT cell viability assays. Furthermore, we used western blotting to evaluate the effect of PDGFR knockdown on the phosphorylation of ERK stimulated with fresh-PRP and FD-PRP. RESULTS Platelet counts in both the fresh-PRP and FD-PRP samples were approximately 10-fold higher than in peripheral blood samples. The phosphorylation and activation of the PDGFR and ERK were evenly induced by fresh-PRP and FD-PRP stimulation. Both freshPRP and FD-PRP significantly induced osteoblast proliferation in MTT cell viability assays. Furthermore, osteoblast PDGFR knockdown attenuated the downstream ERK activation by fresh PRP and FD-PRP. CONCLUSIONS We demonstrated the pharmacological activity of PDGF in FD-PRP in vitro after 4 weeks of storage.