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Exogenous Nitric Oxide Induced Early Mineralization in Rat Bone Marrow Mesenchymal Stem Cells via Activation of Alkaline Phosphatase
Mohammad Hussein Abnosi , Sadieeh Pari
Abstract:  
Background: Since it was shown that low concentration and short time treatment with sodium nitroprusside (SNP), a nitric oxide (NO)–donor, cause no harm to rat bone marrow mesenchymal stem cells (MSCs), we studied the impact of SNP on MSCs differentiation. Methods: MSCs were treated with 100 and 1000 µM of SNP for 1 hour in every 48 hours and after 5, 10, 15 and 21 days in osteogenic media. The viability and the level of mineralization were determined using MTT assay and alizarin red staining, respectively. Morphology of the cells was studied using fluorescent dye. Concentration of calcium and the activity of alanine transaminase (ALT), aspartate transaminase (AST), lactate dehydrogenase (LDH) and alkaline phosphatase (ALP) were evaluated by commercial kits. Results: SNP with the concentration of 1000 µM significantly reduced viability from day 5 to day 20, but 100 µM did not affect the viability until the day 15. The low concentration of SNP increased the matrix deposition from day 10 and reached almost its maximum (4.40 ± 2.4) at the day 15. Also, the increasing activity of ALP (419 ± 2.2) due to low concentration of SNP started at day 10 and continued till the day 20, while LDH (2026 ± 11) and AST (25.6 ± 0.4) elevation was observed from day 5 onwards. In case of ALT we observed a significant decrease (36%) from day 5 till the day 20. Conclusion: Based on our findings, low concentrations of SNP might be useful in the promotion of bone repair.
Keywords: Alkaline phosphatase, Mesenchymal stem cells, Nitroprusside, Osteoblasts
Full-Text [PDF 875 kb]      
Type of Study: Full Length | Subject: Related Fields
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