Mineralization induced by β-glycerophosphate in cultures leads to a marked increase in collagenase synthesis by mouse osteogenic MC3T3-E1 cells under subsequent stimulation with heparin

Sakamoto, Seizaburo; Sakamoto, Masako; Goldberg, Laura; Colarusso, Lucia; Gotoh, Y.

doi:10.1016/0006-291x(89)92377-2

Cited by 29 publications

(16 citation statements)

References 14 publications

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“…4). Our finding that monophosphoesters are better substrates of mineralization is consistent with the several reports on the role of β-GP in stimulating mineralization in osteoblast or chondrocyte in vitro cell cultures (22-26). This finding also draws further support from a kinetic study that demonstrated AMP and β-GP to be better substrates of alkaline phosphatase activity than ATP or ADP (Vmax values of β-GP was 76%, AMP was 62% and ATP was 15% relative to p-nitrophenylphosphate) (27).…”

Section: Resultssupporting

confidence: 93%

A simple and non-radioactive technique to study the effect of monophosphoesters on matrix vesicle-mediated calcification

Garimella

Sipe

Anderson

2004

Biol. Proced. Online

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A simple and non-radioactive technique based on O-cresolpthalein complexone assay was developed to study in vitro non-radioactive calcium ( 40 Ca) deposition by isolated matrix vesicles. Using this technique, the effect of various phosphoester substrates including ATP, AMP and β-GP on in vitro MV-calcification was studied. O-cresolpthalein complexone assay with non-radioactive calcium demonstrated that AMP or β-GP were more effective in promoting calcium deposition by isolated MVs than ATP. The application of this nonradioactive technique, which is highly sensitive and simple, would offer a useful alternative approach to the routinely used radiometric biomineralization assay which employs radioactive 45 Ca.

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Section: Resultssupporting

confidence: 93%

A simple and non-radioactive technique to study the effect of monophosphoesters on matrix vesicle-mediated calcification

Garimella

Sipe

Anderson

2004

Biol. Proced. Online

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“…Many reports indicate that heparin alone inhibits osteoblast growth or differentiation (Sakamoto et al, 1989;Bhandari et al, 1998;Handschin et al, 2005), apart from one report showing that osteoblast differentiation can be controlled depending on the concentration of heparin (Hausser and Brenner, 2004). Here we showed that heparin significantly enhanced ALP activity and mineralization of MC3T3-E1 cells.…”

Section: Discussionmentioning

confidence: 97%

Oversulfated chondroitin sulfate‐E binds to BMP‐4 and enhances osteoblast differentiation

Miyazaki

Miyauchi²,

Tawada³

et al. 2008

Journal Cellular Physiology

110

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Small leucine-rich proteoglycans, such as biglycan, and their side chain sulfated glycosaminoglycans (GAGs), have been suggested to be involved in bone formation and mineralization processes. The present study was designed to investigate whether chondroitin sulfate (CS), one of the GAG, and its oversulfated structures coupled with bone morphogenetic protein-4 (BMP-4) alter the differentiation and subsequent mineralization of MC3T3-E1 osteoblastic cells. CS-E, one of the oversulfated CS structure, enhanced cell growth, alkaline phosphatase (ALP) activity, collagen deposition, and mineralization whereas heparin enhanced only ALP activity and mineralization. As well as CS-E, CS-H, and CPS also enhanced the mineralization of the cells. CS-E enhanced the mineralization of the cells by interacting with protein in the conditioned medium. CS-E induced mineralization was significantly inhibited by an antibody against BMP-4. The addition of exogenous BMP-4 further increased the capacity of CS-E to enhance mineralization. Fluorescence correlation spectroscopy method using fluoresceinamine-labeled GAG revealed that the oversulfated GAGs have a high affinity for BMP-4. The disaccharide analysis of the cells indicated that MC3T3-E1 cells are capable of producing oversulfated structures of CS by themselves. The lack of CS from the cells after chondroitinase treatment resulted in the inhibition of mineralization. These results in the present study indicate that oversulfated CS, which possesses 4,6-disulfates in N-acetyl-galactosamine, binds to BMP-4 and promotes osteoblast differentiation and subsequent mineralization.

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“…In vitro, it potentiates the effect of PTH on mouse calvarial cultures [20] and increases bone resorption by isolated rat and chick [21,22] OC. Of great interest is the fact that heparin stimulates collagenase activity in osteoblastic cell lines [23]; TETs inhibit the same enzyme [24]. The evidence that tetracyclines inhibit bone resorption by interfering with the action of collagenase is circumstantial, but the fact that the augmentation of OC action by heparin is also abolished renders this mechanism more plausible.…”

Section: Discussionmentioning

confidence: 99%

Effect of tetracyclines which have metalloproteinase inhibitory capacity on basal and heparin-stimulated bone resorption by chick osteoclasts

et al. 1993

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Several tetracyclines (TETs) are potent inhibitors of collagenase (CGase) and can inhibit connective tissue degradation in a variety of inflammatory and degenerative disorders. The role of CGase in bone resorption by osteoclasts (OC) remains unclear. Disaggregated OCs from chick embryos were cultured for 24 h on devitalized bovine cortical bone +/- heparin in the presence of various TETs. Doxycycline (Dox) inhibited pit formation in a dose-dependent manner. CMT, a TET derivative which inhibits matrix metalloproteinases (MMPs) but is not antimicrobial, also inhibited chick OC bone resorption. Heparin markedly stimulated bone resorption at 5 micrograms/ml, which was reversed by Dox, 5 micrograms/ml. TETs can reversibly inhibit both basal and heparin-stimulated bone resorption by chick OCs. These findings suggest that MMPs may play a role in osteoclastic bone resorption, and that safe and effective inhibitors of MMPs, including certain TETs, might have a potential therapeutic role.

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Mineralization induced by β-glycerophosphate in cultures leads to a marked increase in collagenase synthesis by mouse osteogenic MC3T3-E1 cells under subsequent stimulation with heparin

Cited by 29 publications

References 14 publications

A simple and non-radioactive technique to study the effect of monophosphoesters on matrix vesicle-mediated calcification

A simple and non-radioactive technique to study the effect of monophosphoesters on matrix vesicle-mediated calcification

Oversulfated chondroitin sulfate‐E binds to BMP‐4 and enhances osteoblast differentiation

Effect of tetracyclines which have metalloproteinase inhibitory capacity on basal and heparin-stimulated bone resorption by chick osteoclasts

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