TERATOGENIC SCREENING IN ZEBRAFISH (Danio rerio) OF THE CRUDE EXTRACT RICH IN CAROTENOIDS OF CANTALOUPE MELON (Cucumis melo L.) AND NANONOENCAPSULATED IN GELATIN
Cucurbitaceae, β-carotene Nanoencapsulation, Embryotoxicity.
Cantaloupe melon has an orange pulp rich in carotenoids, which can promote several beneficial effects on human health. However, carotenoids are extremely unstable in the presence of oxygen, light, and heat, which may reduce their bioactive properties. In this context, nanoencapsulation is an excellent strategy to ensure the preservation and enhancement of bioactive effects. On the other hand, one of the major current concerns is related to the toxicity effects of these nanoparticles. Therefore, this study aimed to evaluate the teratogenicity of the crude extract rich in carotenoids from Cantaloupe melon (CE) and of the porcine gelatin-based nanoparticles containing CE (EPG) in an animal model of zebrafish (Danio rerio). The CE was obtained from Cantaloupe melon (Cucumis melo L.) in processes that involved drying the melon pulp (55 ̊C/24 h) to obtain flour, maceration in ethanol (1:4 w/v) and partition in hexane (1:1 v/v). The nanoparticles were obtained using the oil-in-water (O/W) emulsification technique. For the characterization of nanoparticles in terms of morphology, chemical interactions, particle size and encapsulation efficiency, Scanning Electron Microscopy (SEM), Fourier Transform Infrared Spectroscopy (FTIR), Laser Diffraction and Incorporation Efficiency (IE) analyses were carried out (IE), respectively. In the experiments in an animal model with zebrafish, for the embryotoxicity test, the 308 embryos, divided into seven groups, were exposed to concentrations of 12.5 mg/L and 50.0 mg/L of CE and EPG during 96 hours post fertilization (hpf) and seven days post fertilization (dpf) the optomotor test was performed. In the toxicity tests with adult zebrafish, the 60 fish were divided into six groups exposed to 12.5 mg/L and 50.0 mg/L of CE and EPG for 96 hours, and then submitted to light/dark neurotoxicity, new and social tank. Based on the characterization results, EPG showed an IE of 94% (4.04), a smooth surface without depressions, a diameter of 88.7 nm (7.02), and a polydispersion index of 0.41 (0.03). FTIR evidenced the emergence of new vibrational bands in EPG compared to CE, demonstrating new chemical interactions. In animal model experiments, no anomalies were observed after exposure to 96 hpf, in the groups treated with CE and EPG, and heartbeats remained between 132 and 138 rpm within the expected range for embryos, similar to the negative control and DMSO groups. The groups that received CE and EPG did not show significant morphological alterations. The mortality level was below 20%, revealing neither an embryotoxic nor teratogenic character for CE and EPG in the concentrations used. There was a significant improvement in the visual motor response of larvae exposed to CE and EPG, which can be attributed to the antioxidant power and vitamin A precursor of carotenoids. No adverse effects were identified on the behavior of adults, all were similar to the behavior of the control group, without signs of anxiety, stress, or changes in swimming, speed or sociability that would indicate any toxicity. Therefore, it was found that CE and EPG maintained characteristics found in previous studies, proving the method's reproducibility. There were no signs of teratogenicity, cardiotoxicity, or neurotoxicity in the larvae, and both CE and EPG improved cognitive function and response in the evaluated model. Adult fish showed no signs of anxiety or any other evidence of toxicity, showing that CE and EPG are potentially safe.