Research Article

Artificial Extracellular Matrices with Oversulfated Glycosaminoglycan Derivatives Promote the Differentiation of Osteoblast-Precursor Cells and Premature Osteoblasts

Figure 2

Influence of aECM on TNAP activity, calcium deposition, and morphology of hBMSC. 7,000 hBMSC were plated in BM on aECM. At day 4 after plating, BM was replaced with OM/D. At day 11 after plating, TNAP activity was determined in cell lysates with p-nitrophenylphosphate as a substrate (a). The released p-nitrophenolate was measured photometrically at 405 nm. TNAP activity in mU/mL was normalized to protein concentration in mg/mL determined with Rotiquant assay. At day 22 after plating, calcium deposition was quantified with cresolphthalein complexone at 570 nm (b). Significant differences of col/sGAG-aECM versus col-aECM were calculated by one-way ANOVA analysis and indicated with c ( ), . (c) At day 11 after plating, cells were stained for F-actin fibres with Alexa488-phallodin (green fluorescence) and for nuclei with DAPI (blue fluorescence), scale bar = 50 μm.