Characterization of in vitro osteogenic differentiation of osteoblasts isolated from HO and normal bone.
(A) Human osteoblasts were harvested from excised HO (HO-Ob) and surrounding normal bone (Ob). HO derived osteoblasts showed more ALP activity and more in-vitro bone deposition by Alizarin red staining than cells derived from normal bone. Representative wells and 10x micrographs are shown. (B) Gene transcript levels of Ocn, Osx, and Runx2. (C) Western blot analysis of pSmad 5 protein levels in these cells after 0, 6, and 18 days in osteogenic media. Data shown are reported as the mean ± SD of at least three separate experimental samples performed in triplicate. *p<0.05; **p<0.01. Student T-test.