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Using Alizarin Red Staining to Detect Chemically Induced Bone Loss in Zebrafish Larvae
Published on: December 28, 2021
Regular Supplementation with Antioxidants Rescues Doxorubicin-Induced Bone Deformities and Mineralization Delay in
Sunil Poudel1,2,3, Gil Martins1,2,3, M Leonor Cancela1,2,4
1Centre of Marine Sciences, University of Algarve, 8005-139 Faro, Portugal.
Abstract:
Osteoporosis is characterized by an abnormal bone structure with low bone mass and degradation of microarchitecture. Oxidative stress induces imbalances in osteoblast and osteoclast activity, leading to bone degradation, a primary cause of secondary osteoporosis. Doxorubicin (DOX) is a widely used chemotherapy drug for treating cancer, known to induce secondary osteoporosis. The mechanism underlying DOX-induced bone loss is still not fully understood, but one of the relevant mechanisms is through a massive accumulation of reactive oxygen and nitrogen species (i.e., ROS and NOS) leading to oxidative stress. We investigated the effects of antioxidants Resveratrol and MitoTEMPO on DOX-induced bone impairment using the zebrafish model. DOX was shown to increase mortality, promote skeletal deformities, induce alterations on intestinal villi, impair growth and mineralization and significantly downregulate osteoblast differentiation markers osteocalcin 2 and osterix/sp7. Lipid peroxidation was significantly increased in DOX-supplemented groups as compared to control and antioxidants, suggesting ROS formation as one of the key factors for DOX-induced bone loss. Furthermore, DOX affected mineral contents, suggesting an altered mineral metabolism. However, upon supplementation with antioxidants, DOX-induced effects on mineral content were rescued. Our data show that supplementation with antioxidants effectively improves the overall growth and mineralization in zebrafish and counteracts DOX-induced bone anomalies.
Insights
Antioxidants Resveratrol and MitoTEMPO effectively counteracted bone loss and skeletal deformities caused by the chemotherapy drug Doxorubicin in zebrafish. These findings highlight the role of oxidative stress in Doxorubicin-induced osteoporosis.
Area of Science:
- Bone Biology and Pharmacology
- Zebrafish Models in Toxicology
- Oxidative Stress and Disease Mechanisms
Background:
- Osteoporosis involves bone structure degradation and low bone mass, often linked to oxidative stress.
- Chemotherapy drug Doxorubicin (DOX) can induce secondary osteoporosis through reactive oxygen and nitrogen species (ROS/NOS) accumulation.
- The precise mechanisms of DOX-induced bone loss require further elucidation.
Purpose of the Study:
- To investigate the protective effects of antioxidants Resveratrol and MitoTEMPO against DOX-induced bone impairment in a zebrafish model.
- To explore the role of oxidative stress in DOX-induced bone anomalies.
Main Methods:
- Utilized the zebrafish model to assess Doxorubicin's effects on bone health.
- Administered antioxidants Resveratrol and MitoTEMPO to evaluate their impact on DOX-induced bone impairment.
- Measured mortality, skeletal deformities, intestinal villi alterations, growth, mineralization, osteoblast differentiation markers, lipid peroxidation, and mineral content.
Main Results:
- Doxorubicin treatment increased mortality, caused skeletal deformities, intestinal alterations, impaired growth and mineralization, and reduced osteoblast markers (osteocalcin 2, osterix/sp7).
- Increased lipid peroxidation in DOX groups indicated ROS formation as a key factor in bone loss.
- DOX altered mineral content, but antioxidants rescued these effects, improving overall growth and mineralization.
Conclusions:
- Antioxidant supplementation with Resveratrol and MitoTEMPO effectively mitigates Doxorubicin-induced bone anomalies in zebrafish.
- The study confirms the significant role of oxidative stress in Doxorubicin-induced bone loss.
- Antioxidants show promise in counteracting chemotherapy-induced bone impairment.
