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Cells cultured in osteogenic medium for 6 weeks were assayed using the VK staining method as described previously [23, 24]. In brief, cells were fixed with 4% paraformaldehyde for 30 min at room temperature and then incubated with 5% silver nitrate solution for 1 h by exposure to a 50-W UV lamp. Unincorporated silver nitrate was removed using 5% sodium thiosulfate and rinsed three times with distilled water.
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