Biotechnological Valorization of Grape Pomace: Use of the Zebrafish Model to Evaluate the Toxicity and Teratogenic Effects of Anticancer Biocompounds

Abstract

Optimization of enzyme-assisted polyphenol extraction reduced cellulase use by 7.7 times compared to that reported in the literature. In this way, an extract with 25% more polyphenols and a higher antioxidant capacity was obtained compared to the control. The main compounds identified were (+)-catechin, (-)-epicatechin and rutin. Toxicological analysis using the zebrafish model established a No-Observed-Adverse-Effect Level (NOAEL) of 0.2 mg/mL and an LC50 of 0.46 mg/mL. The extract showed inhibitory effects on lung and liver cancer cells, with IC50 values of 0.18 and 0.19 mg/mL, respectively, as well as effects on pancreatic and colon cells. However, the Lowest-Observed-Adverse-Effect Level (LOAEL) was 0.4 mg/mL, exhibiting dose-dependent embryotoxicity and teratogenesis. These findings highlight the importance of conducting thorough toxicological studies on biocompounds derived from agro-industrial by-products to ensure their viability and safety in therapeutic and food applications.