Characterization of porcine dentin sialoprotein (DSP) and dentin sialophosphoprotein (DSPP) cDNA clones

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/74726/1/j.1600-0722.2003.00009.x.pd

Porcine Enamel Protein Fractions Contain Transforming Growth Factor‐β1

Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/141064/1/jper1688.pd

Simvastatin suppresses the differentiation of C2C12 myoblast cells via a Rac pathway.

Statins, which are known as cholesterol-lowering drugs, have several additional effects including the enhancement of bone formation and the stimulation of smooth muscle cell proliferation. In this study, we investigated the signal pathway of simvastatin operating in C2C12 myoblast cells. Myotube formation of C2C12 cells was efficiently blocked by 1 muM simvastatin, and mevalonic acid was able to cancel this effect. Geranylgeranyl pyrophosphate restored the myotube formation, whereas farnesyl pyrophosphate did not. These findings demonstrate that the Rho family, such as Rho, Rac and Cdc42, occurring downstream of geranylgeranyl pyrophosphate in the mevalonic acid pathway, was involved in the simvastatin-mediated blockage of myotube formation. An inhibitor of Rho kinase did not influence the myotube formation; whereas an inhibitor of Rac blocked this process. Taken together, we conclude that the differentiation of C2C12 cells into myotubes was blocked by simvastatin through the pathway mediated by Rac, not by Rho

Liver-type of tissue non-specific alkaline phosphatase is induced during mouse bone and tooth cell differentiation

Background and objective: Tissue non-specific alkaline phosphatase (TNSALP) contains two types?bone- and liver-type?which are produced from the same gene due to differences in splicing. These two differ in their promoter, but the amino acid sequences of the mature proteins are identical. In this study, we examined the relationship between the two types of TNSALP expression and osteoblast differentiation. Design: Gene expression of the two types of TNSALP was observed by reverse transcription-polymerase chain reaction. MC3T3-NM4 was sub-cloned from an established mouse osteoblastic cell line in which osteoblast characters do not appear without dexamethasone. The C2C12 mouse myoblastic cell line, which can be induced to osteoblasts with bone morphogenic protein 2, and organ-cultured tooth germs were also used in this work. Results: The gene expression of liver-type TNSALP was observed in only MC3T3-NM4 activated by dexamethasone. For C2C12, the gene expression of bone-type TNSALP was observed even in non-induced conditions where myotubes were formed, whereas the liver-type TNSALP mRNA was only expressed when C2C12 differentiated into osteoblasts by bone morphogenic protein 2. Furthermore, in the organ-cultured tooth germs, the liver-type TNSALP mRNA was expressed according to differentiation of tooth germs. Conclusion: These results suggest that the liver-type TNSALP mRNA is induced according to differentiation of bone and tooth

IMMUNOLOGICAL DIFFERENTIATION OF HUMAN TISSUE-NONSPECIFIC TYPE ALKALINE PHOSPHATASES BY A MONOCLONAL ANTIBODY TO THE ENZYME OF HUMAN OSTEOBLAST-LIKE CELLS

Monoclonal antibodies against alkaline phosphatase [ALP; ortho-phosphoric monoester phosphohydrolase, alkaline optimum, EC 3.1.3.1.] of cultured human osteoblast-like cells (HBC) were raised in mice. Immuno-reactions of tissue-nonspecific type ALP from human bone, dental pulp, liver and kidney as well as intestinal and placental types to the monoclonal antibodies were compared by a dot immunoassay and ELISA. One clone was able to recognize antigenic differences among tissue-nonspecific type ALPs in addition to intestinal and placental ALPs; it reacted favorably with ALPs from HBC, human bone, kidney and dental pulp, but not with human liver enzyme. Similarly, the antibody immunoreacted with bone-derived ALP but not with liver-derived enzyme present in human serum.The present monoclonal antibody preparation can be utilized in basic studies as well as in clinical laboratory tests to distinguish minor heterogeneity among human Al Ps

VOLUNTARY EXERCISE INCREASES OSTEOGENETIC ACTIVITY IN RAT BONES

The purpose of this study was to investigate the effect of voluntary exercise on osteoinductive activity in rat bone. Sprague-Dawley male and female rats were allowed to exercise freely by running on a treadmill or kept as controls without exercise for 53 days. Decalcified humeral diaphyses from experimental and control rats were implanted intraperitonealy into host rats and harvested after 33 days. A significant increase in bone formation was confirmed in the implanted bone matrices from the running group in comparison with those from control animals by soft X-ray photography and determination of alkaline phosphatase activity and mineral content. Alkaline phosphatase activity in bone and serum was increased by exercise in both male and female animals. The results suggest that osteoinductive activity in the bone was probably due to increased levels of bone morphogenetic protein following voluntary exercise

Direct action of nitric oxide on osteoblastic differentiation

AbstractThe effect of nitric oxide (NO) on osteoblastic differentiation was examined in cultured mouse osteoblasts. Interleukin-1β and tumor necrosis factor-α expressed inducible NO synthase gene with little effect on constitutive NO synthase gene. These cytokines increased NO production, which was inhibited by l-NMMA pretreatment, and decreased alkaline phosphatase (AIPase) activity, which was not restored by l-NMMA. Furthermore, NO donors, sodium nitroprusside and NONOate dose-dependently elevated AIPase activity and expression of osteocalcin gene. These results suggest that NO directly facilitates osteoblastic differentiation and the cytokine-induced inhibition of AIPase activity is mediated via mechanism other than NO