Cytological Effects of Serum Isolated from Polytraumatized Patients on Human Bone Marrow-Derived Mesenchymal Stem Cells

Figure 6

Osteogenic differentiation potential. (a) Alizarin red staining showed calcium deposits as red staining after culturing hBMSCs in osteogenic differentiation medium containing 10% serum from HS, D1, D5, D10, or FCS for 14 days. (a1) Macroscopic image of alizarin red staining. (a2) Cells prior to alizarin red staining at 40x magnification. (a3) Images showing the hBMSCs at a 40x magnification after staining with alizarin red. (a4) Analysis of alizarin red staining of calcium deposits using a tool based on OpenCV library (version 4.1.0). Areas without staining were color isolated as black areas. The pixel ratio of the area of interest (nonblack area) in comparison to the black area was obtained. In the groups containing human serum (HS, D1, D5, and D10), a larger area of the 6-well was positive for alizarin red staining than in the FCS group (). (b) Quantitative analysis of the amount of alizarin red staining quantified using a software tool to compare the relative areas positive for staining. D1 showed significantly more area of alizarin red staining when compared to the FCS group. (). (c) Analysis of alizarin red staining indicative of osteogenic differentiation with higher concentrations of calcium deposition after 14 days of culture. HS, D1, and D10 showed significantly higher absorbance as compared to the FCS group. ().

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