Avaliação do efeito de uma lectina isolada de ovos do gastrópode marinho Aplysia dactylomela na cicatrização de feridas induzidas cirurgicamente em camundongos

Abstract

The wounds result from the rupture of the skin associated with damaging effects on the structure that integrates the extracellular matrix. Once damaged, the skin loses its functions and can compromise people's physical and mental health, and can even lead to death. Hundreds of bioactive compounds have already prospected from invertebrates. Among these compounds are lectins, proteins that recognize and bind specific carbohydrates and can be found on cell surfaces, playing an important role in cell interaction and communication. The present study evaluated the cytotoxic and healing effect of lectin isolated from eggs of the marine gastropod Aplysia dactylomela (ADEL) in experimental models in vitro and in vivo. In order to evaluate the cytotoxicity of ADEL on murine fibroblasts (L929) and human keratinocytes (HaCat), the cell viability assay was performed by MTS. The adult zebrafish animal model (Danio rerio) was used to assess acute toxicity and the effect on locomotor activity. For analysis of pro-healing activity, surgical wounds of 6 mm in diameter were induced in the dorsal region of Swiss mice (n = 60) that were treated with a topical daily application of 100 μL according to the test group (ADEL 125 μg / mL ; dADEL 125 μg / mL; BSA 125 μg / mL; NaCl 150mM; Vitamin C 125 μg / mL) for 12 days. The wounds were evaluated taking into account the following parameters, edema, hyperemia, exudation, presence of crust and bleeding. In addition, for microscopic analysis, tissue samples corresponding to the area of the experimental lesion on the 3rd, 7th and 12th postoperative days were biopsied. The cytotoxic results obtained showed that fibroblasts treated with ADEL at 24h, 48h and 72h remained viable above 80% in all tested concentrations (7.8 - 500 μg / mL), the same was observed for keratinocytes . In the locomotor activity of zebrafish exposed to concentrations of 125 μg / mL, 250 μg / mL and 500 μg / mL of ADEL, neither acute toxicity was verified in any of the doses tested for up to 96 hours after exposure to ADEL in relation to the control ( p <0.05). Skin wounds treated with ADEL showed more discrete clinical signs such as hyperemia, edema, and exudate, with total re-epithelialization in the 12th postoperative period, the same was observed in the other experimental groups. In P.O 12, the groups treated with ADEL and dADEL presented well-structured and adequate skin layers, however the dADEL group with greater traction. In conclusion, ADEL contributed to the earlier healing, without showing cytotoxic effects, with a discreet action on tissue repair.