Long Chain Omega-3 Polyunsaturated Fatty Acid Supplementation Protects Against Adriamycin and Cyclophosphamide Chemotherapy-Induced Bone Marrow Damage in Female Rats

Figures

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  Figure 1. Effects of daily omega-3 polyunsaturated fatty acid (PUFA) supplementation and 4 weekly AC treatment on daily body weights. Vertical arrows indicate days of injections with water control or adriamycin and cyclophosphamide in combination (AC).

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  Figure 2. Effects of daily omega-3 polyunsaturated fatty acid (PUFA) supplementation during weekly AC treatment on bone marrow constitution. Treatment effects on counts of bone marrow non-adherent cells isolated from treated rats (A); Haematoxylin and eosin (H&E)-stained sections of the lower secondary spongiosa region of the tibia from a control rat (B); an AC-treated rat (C) and an AC + PUFA-treated rat (D); with yellow arrows pointing to non-fat cell bone marrow cells, red arrows pointing to adipocytes. Scale bar on all panels = 200 µm. Treatment effects on the marrow cellularity (%) (E) and marrow adiposity % (F). Significant values are indicated as: * p < 0.05 and ** p < 0.01 compared to control.

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  Figure 3. Effects of daily omega-3 polyunsaturated fatty acid (3PUFA) supplementation during weekly AC treatment on metaphyseal trabecular bone volume and structure. μ-CT longitudinal cross-section of a control rat tibia (A); Region within the red dotted line (region of interest) represents the first 2 mm of metaphysis from the end of growth plate, where all μ-CT scan data were analysed. Analyses of the treatment effects on trabecular bone volume/tissue volume (BV/TV) (B) and trabecular structures including trabecular number (C); trabecular thickness (D) and trabecular spacing (E).

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  Figure 4. Effects of daily omega-3 polyunsaturated fatty acid (3PUFA) supplementation during weekly AC treatment on bone cells at primary spongiosa. H&E stained metaphysis of a control rat (A); with arrows pointing to bone lining cells. Analyses of the treatment effects on bone lining cell density (B); Tartrate-resistant acid phosphatase (TRAP)-stained tibia sections of an AC-treated rat (C) and AC + PUFA-treated rat (D); with arrows pointing to multinucleated osteoclasts. Treatment effects on osteoclast density (E). Scale bar on all panels A, C and D = 50 µm.

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  Figure 5. Effects of daily omega-3 polyunsaturated fatty acid (PUFA) supplementation during weekly AC treatment on ex vivo osteoclast formation potential. Ex vivo osteoclast formation from bone marrow cells of an AC-treated rat (A) and from bone marrow cells of AC + PUFA-treated rat (B); with arrows pointing to multinucleated tartrate-resistant acid phosphatase-stained osteoclasts. Treatment effects on the ex vivo osteoclast formation ability (C); size of osteoclasts formed (D); and numbers of nuclei within osteoclasts formed (E) (* or ** compared to the respective group with 3–5 nuclei). Scale bar on all panels = 100 µm. # above horizontal bars reflects differences between connected groups. Significant values are indicated as: * or # = p < 0.05 and ** = p < 0.01.

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  Figure 6. Long chain omega-3 polyunsaturated fatty acid (PUFA) supplementation helps to preserve bone and bone marrow microenvironment during adriamycin and cyclophosphamide (AC) combination chemotherapy. It protects against AC treatment-induced bone marrow cell depletion and adiposity and attenuates separation of bone trabeculae in rats and osteoclast formation ex vivo.