Osteogenic Potential of Fibromodulin in mSSC

FACS-isolated mSSC were harvested from DO Den and DO Inn specimens at POD 10 and cultured in 96 well plates over a course of 10 days in osteogenic medium with media changes every 48 h in duplicates. Cells were treated with and without a single dose of hFibromodulin (FMOD) protein (0.1 mg/ml, R&D 9840-FM-050) at 12 h to evaluate the ability of FMOD to augment osteogenic potential. The monolayer of cells was fixed with 100% ethanol for 15 min and stained with alizarin red solution for 1 h at room temperature. The cells were washed several times with ultrapure water and imaged for osteogenic potential immediately. Following imaging, the cells were treated with a methanol/acetic acid mixture for 15 min and absorbance was detected at 450 nm using NanoDrop™ 0ne^c Microvolume UV–Vis Spectrophotometer (ThermoFisher)^{12 (link)}.