Fig. 2: Metabolome analysis reveals nanotopography-mediated changes in cellular respiration, independent of mitochondrial dynamics.

a Stro-1⁺ (red) or total BM (green) MSCs were cultured on SQ or flat surfaces for 7 or 28 days, and the number of metabolites specific or common to both cell types were enumerated. Common metabolites with a confidence value of 10 were identified using IDEOM software in both MSC populations (b), and the heat map shows the distribution of these at day 7 of culture (c). d Fold change in selected metabolite concentrations. Data in (a–d) show mean of 6 nanotopographies per condition. e Biochemical network analysis of metabolite changes in MSCs cultured on flat versus SQ. f Changes in mitochondrial activity were measured using JC-1 staining, in MSCs cultured on SQ or NSQ nanotopographies relative to flat control. Each data point is the mean of 3 technical repeats per independent donor (n = 4). Comparisons by one-way ANOVA (*p = 0.0079). g, h Mitochondrial mass (Mitotracker Green) and superoxide generation (Mitosox Red) were also evaluated by flow cytometry (single measurement from n = 4 donors). i Stro-1⁺ MSCs were cultured on flat and SQ nanotopographies (n = 4 topographies per donor, n = 4 independent donors) for 14 days in the presence (hatched bars) or absence (open bars) of the ROCK inhibitor, Y27632, and changes in mitochondrial activity measured using JC-1 staining. Means ± SEM are shown in f, g, h and i and number of donors (N) are shown for each condition. Means ± S.D. in d, g and h by one-way ANOVA. Direct comparisons of means by two-tailed student T-test (Mann–Whitney), *p < 0.05; n.s., non-significant). Source data are provided as a Source data file.