Figure 3From: Soluble factors from biofilms of wound pathogens modulate human bone marrow-derived stromal cell differentiation, migration, angiogenesis, and cytokine secretion Soluble factors from biofilms differentially affect adipogenic and osteogenic differentiation of hBMSCs in vitro. A) Light microscopy images of hBMSCs stained with Oil Red O (red, 40x magnification) after adipogenic differentiation on days 7, 14 and 21 in the presence of BCM (25%) from S. aureus or P. aeruginosa. B) Quantification of Oil Red O stain (OD500nm) to detect intracellular oil droplets following adipogenic differentiation determined by extraction and solubilization of the dye. C) Western blot analysis of adipogenic proteins: adiponectin, peroxisome proliferator-activated receptor gamma (PPARγ), and leptin. D) Light microscopy images of hBMSCs stained with Alizarin Red S (red, 40x magnification) after osteogenic differentiation on days 7, 14 and 21 in the presence of BCM (25%) from S. aureus or P. aeruginosa. E) Quantification of Alizarin Red S stain (OD415nm) to detect intracellular calcium deposits following osteogenic differentiation. F) Western blot analysis of osteogenic proteins: Runt-related transcription factor 2 (RUNX2), bone gamma-carboxyglutamic acid-containing protein (osteocalcin), and alkaline phosphatase. Quantification results are expressed as a ratio of non-differentiated growth controls (dashed line) at each timepoint; *p < 0.05 compared to differentiation control at each respective timepoint.Back to article page