Platelet-rich plasma stimulates osteoblastic differentiation in the presence of BMPs

https://doi.org/10.1016/j.bbrc.2007.06.142Get rights and content

Abstract

Platelet-rich plasma (PRP) is clinically used as an autologous blood product to stimulate bone formation in vivo. In the present study, we examined the effects of PRP on proliferation and osteoblast differentiation in vitro in the presence of bone morphogenetic proteins (BMPs). PRP and its soluble fraction stimulated osteoblastic differentiation of myoblasts and osteoblastic cells in the presence of BMP-2, BMP-4, BMP-6 or BMP-7. The soluble PRP fraction stimulated osteoblastic differentiation in 3D cultures using scaffolds made of collagen or hydroxyapatite. Moreover, heparin-binding fractions obtained from serum also stimulated osteoblastic differentiation in the presence of BMP-4. These results suggested that platelets contain not only growth factors for proliferation but also novel potentiator(s) for BMP-dependent osteoblastic differentiation.

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Materials and methods

Materials. Bovine plasma was purchased from Funakoshi, Tokyo, Japan. Dulbecco’s modified Eagle’s medium (DMEM) α-minimal essential medium (α-MEM) were obtained from Sigma–Aldrich (St. Louis, MO). Fetal bovine serum (FBS) and calf serum (CS) were obtained from Moregate Biotech (Bulimba, Qld, Australia). Bovine thrombin was obtained from Wako Pure Chemical Industries (Osaka, Japan). Recombinant BMP-2 produced in Escherichia coli was obtained from Acris Antibodies GmbH (Nordhein-Westfalen,

PRP and soluble fraction of PRP stimulate BMP-4-induced osteoblastic differentiation and proliferation of C2C12 myoblasts

BMP-4 at 20 ng/ml slightly induced ALP activity in C2C12 cells (Fig. 1A). Addition of human PRP to the cultures stimulated ALP activity at lower concentrations, although the degree of stimulation gradually decreased at higher concentrations (Fig. 1A). In contrast, human PPP failed to stimulate ALP activity at all concentrations examined (Fig. 1A). We further examined the stimulatory activity in the soluble fractions prepared from bovine activated PRP and PPP. The ALP activity induced by BMP-4 at

Discussion

In the present study, we found that the soluble fraction prepared from activated PRP stimulates osteoblastic differentiation in the presence of BMPs in mesenchymal progenitor cells. Pretreatment of scaffolds with BMP-4 and the soluble PRP fraction stimulated osteoblastic differentiation in vitro. Because PRP is an autologous blood product, simultaneous application of PRP and BMPs with scaffolds may be a simple and useful method for enhancement of bone formation in vivo.

The stimulatory activity

Acknowledgments

This work was supported in part by the Ministry of Education, Culture, Sports, Science and Technology of Japan, and in particular by a Ministry Grant to the Saitama Medical University Research Center for Genomic Medicine.

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