The effect of combined application of TGFp-1, BMP-2, and COLLOSS~R E on the development of bone marrow derived osteoblast-like cells in vitro

摘要

This study investigated the combined application of Transforming Growth Factor beta-1 (TGF beta-1) and Bone Mor-phogenetic Protein-2 (BMP-2) to stimulate osteogenic expression in vitro. TGFp-1 and BMP-2 fulfill specific roles in the formation of new bone. COLLOSS~R E, a bone-derived collagen product containing a variety of naturally occurring growth factors, was also used. Growth factors were administered to osteoblast-like cells from rat bone marrow (RBM). Proliferation and differentiation were monitored up to 24 days, by measuring total DNA content, alkaline phosphatase activity, and calcium content. Genetic expression of a set of differentiation markers at day 7 was measured by Q-PCR. Adding BMP-2 alone induced high proliferation rates, compared to the growth factor supplemented groups, and it induced high differentiation rates, compared to the control group. Adding TGFp-1 combined with BMP-2, TGFp-1 alone, or COLLOSS~R E resulted in a significant decrease in proliferation rate, but an increase in differentiation rate, compared to the control group. Additive or synergistic effects of ' application of TGF beta-1 and BMP-2 were not observed. The observed effects of COLLOSS~R E mainly resembled those of TGF beta-1 application alone. It can be concluded that BMP-2 is the most suitable candidate for osteogenic stimulation of RBM cells in these settings.