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Effect of the bone resorption supernatant from RAW264.7 osteoclast on the osteogenic activity of mouse MC3T3-E1 cell

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Author:
No author available
Journal Title:
Chinese Journal of Stomatology
Issue:
1
DOI:
10.3760/cma.j.issn.1002-0098.2012.01.012
Key Word:
破骨细胞;成骨细胞;骨钙素;抗酒石酸酸性磷酸酶染色;Osteoclast;Osteoblast;Osteocalcin;Tartrate resistant acid phosphatase stain

Abstract: Objective To investigate the effect of osteoclast bone resorption supernatants on the osteogenic activity of mouse MC3T3-E1 cell line.Methods Mouse RAW264.7 cell line was induced to osteoclast which was identified with tartrate resistant acid phosphatase (TRAP) staining and osteoclast specific gene detection.The differentiated RAW264.7 osteoclast was co-cultured with bovine milling bone specimen followed by toluidine blue staining.Then mouse MC3T3-E1 cell was cultured with supernatant from the osteoclast bone absorbent model.Methyl thiazolyl tetrazolium (MTT) method,alizarin red S staining,enzyme-linked immunosorbent assay detection of osteocalcin,and reverse transcriptase polymerase chain reaction detection were adopted to investigate the proliferation,calcification and osteogenic activity of MC3T3-E1 cells.Results TRAP staining,osteoclast specific gene detection and toluidine blue staining all indicated that RAW264.7 cell could be differentiated into functioning osteoclast.The supernatant from the osteoclast bone absorbent model could inhibit the proliferation of MC3T3-E1 cells,with the A value between 0.062 ± 0.004 and 0.405 ± 0.033 (P < 0.05 ).It could also increase the formation of calcification nods,promote the osteocalcin level which peaked with the tenth day's supernatant at a level of (2.965 ±0.047 ) μg/L,as well as enhance the transcription of the alkaline phosphatase and Runt related transcription factor 2 gene.Conclusions RAW264.7 osteoclast bone absorbent supernatant might influence the osteogenic activity of osteoblast-like cell by inhibiting proliferation, promoting differentiation and calcification.

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