In Vivo Genotoxicity Evaluation of a Stilbene Extract Prior to Its Use as a Natural Additive: A Combination of the Micronucleus Test and the Comet Assay.

Concepción Medrano-Padial,María Puerto,Ana Isabel Prieto,Caroline Rouger,Nahúm Ayala,Stéphanie Krisa,Silvia Pichardo,Pauline Beaumont

doi:10.3390/foods10020439

Abstract

Genotoxic data of substances that could be used as food additives are required by the European Food Safety Authority. In this sense, the use of an extract from grapevine shoots containing a stilbene richness of 99% (ST-99), due to its antioxidant and antibacterial activities, has been proposed as an alternative to sulfur dioxide in wine. The aim of this work was to study, for the first time, the in vivo genotoxic effects produced in rats orally exposed to 90, 180, or 360 mg ST-99/kg body weight at 0, 24, and 45 h. The combination of micronucleus assay in bone marrow (OECD 474) and standard (OECD 489) and enzyme-modified comet assay was used to determine the genotoxicity on cells isolated from stomach, liver, and blood of exposed animals. The ST-99 revealed no in vivo genotoxicity. These results were corroborated by analytical studies that confirm the presence of stilbenes and their metabolites in plasma and tissues. Moreover, to complete these findings, a histopathological study was performed under light microscopy in liver and stomach showing only slight modifications in both organs at the highest concentration used. The present work confirms that this extract is not genotoxic presenting a good profile for its potential application as a preservative in the wine industry.

Highlights

Many natural compounds such stilbenes or their analogues have awakened the interest of the scientific community because of their potential use as additives in the food industry [1,2]
No significant difference in the polychromatic erythrocytes (PCE)/NCE and PCE/total erythrocytes ratio between the groups treated with stilbene richness of 99% (ST-99) extract and the negative control group was observed in either sex
The results indicated that the exposure of 90, 180, and 360 mg ST-99/kg b.w. in both sexes did not induce an increase in the frequency of Endo III or FPG-sensitive sitesEnzyme-Modified in any tissue assayed to the control group (Figure 1b, c)

Summary

Introduction

Many natural compounds such stilbenes or their analogues have awakened the interest of the scientific community because of their potential use as additives in the food industry [1,2]. Today’s consumers demand high quality foods that are free from chemical substances, fresh tasting, microbiologically safe, and with an extended shelf-life [5] In this regard, favorable results have been obtained with the use of natural compounds such as stilbenes and flavonoids. Some studies have demonstrated that phenolic compounds naturally found in grape extracts present a high antimicrobial activity against bacteria that cause numerous deterioration in wine [6,7,8]. These compounds exhibited great antioxidant capacity such as scavenging of free radicals [9,10,11,12].

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