Figure 1

Growth and survival. Larvae were fed with resveratrol (RES), MitoTEMPO (MT) and doxorubicin (DOX) alone or in combination for 30 days. Total length of zebrafish larvae at 15 days post fertilization (dpf) (N = 25 × 4) (a) and 30 dpf (N = 25 × 4) (b), Dry weight at 30 dpf (N = 10 × 4) (c) and Survival at 30 dpf (N = 100 × 4) (d). Levels of significance were calculated using Tukey’s multiple comparisons (one-way ANOVA) [* p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001].
Figure 2

Histology of zebrafish gut. Total of 30 dpf zebrafish guts sections was stained with H&E to observe the villi from the different treatment groups with resveratrol (RES), MitoTEMPO (MT) and doxorubicin (DOX) (a). Length of villi (b). Statistical significance was calculated using Tukey’s multiple comparison (one-way ANOVA) [*** p ≤ 0.001, **** p ≤ 0.0001] [N = 5 × 4].
Figure 3

Incidence and distribution of skeletal deformities. Incidence of skeletal deformities (a). Partial Least-Squares Discriminant Analysis (PLS-DA score) on the incidence of deformities of zebrafish supplemented with antioxidants, resveratrol (RES), MitoTEMPO (MT), and pro-oxidant, doxorubicin (DOX) (b). Distribution of skeletal deformities (c) and heatmap of distribution of skeletal deformities (d). Statistical significances were calculated using Tukey’s multiple comparisons (one-way ANOVA) [* p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001] [N = 20 × 4].
Figure 4

Mineralization of zebrafish vertebral column. Percentage of mineralized (red), mineralizing (pink), unmineralized (blue) vertebrae (a) and graphical representation of the mineralization status of the zebrafish. Mineralization status of the vertebrate (b), (red represents mineralized vertebrae and pink represents mineralizing vertebrate). Heat map of mineralization of vertebrae of zebrafish feed with resveratrol (RES), MitoTEMPO (MT) and doxorubicin (DOX) supplemented microdiets (c). Levels of significance were calculated using Tukey’s multiple comparisons (one-way ANOVA) [* p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001] [N = 20 × 4].
Figure 5

Mineral analysis of zebrafish fed with microdiets supplemented with antioxidants, resveratrol (RES) and MitoTEMPO (MT), and pro-oxidant, doxorubicin (DOX). Mineral content of Calcium (a), Phosphorus (b), Ca/P ratio (c), Sodium (d), Potassium (e) and Magnesium (f). Level of significance was calculated using Tukey’s multiple comparisons (one-way ANOVA) [* p ≤ 0.05 and ** p ≤ 0.01] [N = 8 × 4].
Figure 6

Lipid peroxidation on zebrafish fed microdiets enriched with antioxidants, resveratrol (RES) and MitoTEMPO (MT), and pro-oxidant, doxorubicin (DOX). Lipid peroxidation of the zebrafish larvae at 15 dpf (a) and 30 dpf (b). Levels of significance were calculated using Tukey’s multiple comparisons (one-way ANOVA) [* p ≤ 0.05 and ** p ≤ 0.01].
Figure 7

Doxorubicin affects osteoblastic markers. mRNA expression of osteoblast differentiation markers on zebrafish fed with resveratrol (RES), MitoTEMPO (MT) and doxorubicin (DOX) supplemented microdiets, alone or in combination; osteocalcin 2 (oc2) (a), osterix/sp7 (sp7) (b) and runx2b (c). Levels of significance were calculated using Student’s t-test [* p ≤ 0.05] [N = 10 × 4].
Acknowledgments
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