Fig. 7.
MSCs assembled into multicellular clusters and produce more pericellular matrix within microstructured ALG-KHhydrogels. A) Mechanosensing in microstructured hydrogels (Aii) Gene expression of CTGF represented as the fold change in expression levels of ALG-KH-RGD-B in comparison to ALG-KL-RGD-B. (Aii) Expression of YAP marker in MSCs embedded in ALG-KH-RGD hydrogels, showing higher translocation to the nuclei (in pink) in MSCs anchoring to the (a) microstructures when compared to MSCs (b) in the surrounding regions (day 7). B) Effect of the microstructures on the gene expression of osteogenesis and chondrogenesis markers, represented as the fold change in expression levels of ALG-KH-RGD-B in comparison to ALG-KL-RGD-B. C) MSCs spreading in non-degradable (B) versus degradable (OX) hydrogels. (Ci) Actin and nuclei staining showing differences in MSCs spreading ability in smooth hydrogels (ALG-KL-RGD-B), hydrogels with microstructural domains (ALG-KH-RGD-B) or combined with degradable (ALG-KH-RGD-OX) hydrogels. (Cii) Quantification of total cell area and degree of cell-cell interconnectivity (Ciii) obtained by image analysis. (Civ) MMP activity in conditioned media from MSC-laden ALG-KH-RGD-B and ALG-KH-RGD-OX hydrogels, assessed by gelatin-zymography. D) Gene expression analysis of ECM proteins (FN1 and COL1A1) over time in culture. Bars represent mean values; error bars represent standard deviation (n ≥ 3). E) Differences in fibronectin (FN) production on day 14. Data shown for at least 3 individual experiments. Statistical differences are represented by ∗ (P < 0.05) ∗∗ (P < 0.01), ∗∗∗ (P < 0.001) and ∗∗∗∗ (P < 0.0001). (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)