Bone morphogenetic proteins (BMP) stimulate osteoblast differentiation by signal transduction via three BMP receptors (BMPR-IA, -IB, and -II). Several growth factors, including transforming growth factor-beta1 (TGF-beta1), fibroblast growth factor-2 (FGF-2) and platelet-derived growth factor-AB (PDGF-AB), have also been shown to play an important part in osteogenesis. The mechanism underlying these activities is unclear, but these growth factors could modulate the BMP/BMPR pathway by up-regulating BMPR expression, thereby enhancing the osteogenic responses of bone cells to the BMP. In this study we have therefore examined the effects of TGF-beta1, FGF-2, and PDGF-AB on BMPR expression and BMP-2-mediated osteoblast functions in primary human bone cells. The results showed that although the ligand BMP-2 and growth factors had little effect on BMPR-IA and -II transcript expression, they significantly up-regulated BMPR-IB mRNA specifically. However, only the growth factors, but not the ligand BMP-2, increased the surface expression of the BMPR-IB antigen, which was found to be due to a differential effect of BMP-2 and the growth factors on the Smurf1/Smad6-induced breakdown process. Pre-incubation of the cells with the growth factors significantly augmented BMP-2-induced Smad1/5/8 phosphorylation, and Dlx5 expression ALP activity, compared with that of cells treated with BMP-2 alone. When cells were transfected with siRNA targeting BMPR-IB, the growth factors neither up-regulated BMPR-IB transcript expression nor enhanced BMP-2-induced Smad1/5/8 phosphorylation, Dlx5 expression and ALP activity. The results indicate that increased BMPR-IB by TGF-beta1, FGF-2, and PDGF-AB significantly enhances BMP-2-induced osteogenic functions in vitro, suggesting that they might positively modulate bone formation by up-regulating BMPR-IB in vivo.

Cissus quadrangularis Linn. has been implicated as therapeutic agent for enhancing bone healing. Though its osteogenic activity has been suggested, the underlying mechanism still remains unclear. In the present study, the effects of ethanol extract of C. quadrangularis (CQ-E) on osteoblast differentiation and function were analyzed using murine osteoblastic cells. The results indicated that mRNA expressions of osteoblast-related genes were not affected by the CQ-E treatment. However, alkaline phosphatase (ALP) activity and the extent of mineralized nodules were significantly increased in treated cells compared with controls. The addition of an extracellular regulated kinase 1/2 inhibitor, a Jun N-terminal kinase 1/2/3 inhibitor and a p38 mitogen-activated protein kinase (MAPK) inhibitor resulted in significantly decreased ALP activity, preferentially by p38 MAPK inhibitor. These results suggested that CQ-E may regulate osteoblastic activity by enhancing ALP activity and mineralization process, and the increased ALP activity effect of CQ-E is likely mediated by MAPK-dependent pathway.

Cells derived from the periodontal ligament (PDL) have previously been reported to have stem cell-like characteristics and to play an important part in re-building damaged tissue, including alveolar bone. However, these populations have been heterogeneous, and thus far no highly purified periodontal stem cell (PSC) clone has yet been established from adult human PDL tissue. The present study was therefore carried out to isolate single cell-derived PDL clones and to delineate their phenotypic and functional characteristics. In this report we have obtained four homogeneous and distinct clones--namely, C5, C6, C7, and C8--and have found these to be highly proliferative and to express the stromal cell markers CD29 and CD44. In particular, C7 showed stem cell-like characteristics of small cell size with reduced cytoplasm, clonogenicity, and multilineage potential, including osteogenic activity in forming bone-like tissue in organoid micromass cultures. Clones C5 and C6 possessed osteoprogenitor features with mineralized matrix-forming activity, whereas C8 did not undergo osteogenic, adipogenic, or chondrogenic differentiation. The present study thus reports, for the first time, the isolation and cellular and molecular characterization of highly purified putative PSC and osteoprogenitors in adult human PDL, based on clonogenicity and multilineage differentiation potential, with PSC-C7 capable of bone formation in vitro, suggesting that such cells may have potential value for stem cell-based bone tissue engineering in vivo.