Fig. 8.

Behaviour of HBMSCs on human bone ECM hydrogels. (a) HBMSCs were observed to attach and spread on the ECM hydrogels, irrespective of the hydrogel source from various bone powder sizes (scale bar = 2 μm). (b) ECM hydrogels were placed in the centre of tissue culture plate (TCP), and after seeding HBMSCs, their behaviours were observed using a time-lapse microscope. Some cells from the TCP migrated towards the ECM hydrogels (indicated by red arrows), while cells on the ECM hydrogels spread on top of the hydrogels (indicated by yellow arrows) (scale bar = 50 μm). (c and d) Cells stained with Calcein AM and DiD (green and red, respectively) were cultured in ECM hydrogels for 1, 3, and 7 days. (scale bar = 100 μm) (e) Cell viability and proliferation were evaluated using confocal imaging analysis and the WST-1 assay (n = 5). The cell population was noted to double within 7 days, and there was no significant difference in ECM hydrogels derived from various bone powder sizes. Statistical significance was determined using a one-way test with Tukey's multiple comparisons test (∗p < 0.05 ∗∗p < 0.005 ∗∗∗p < 0.0005∗∗∗∗).(For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article).