Area of Packaging Materials and Systems

Paterna, Spain

Area of Packaging Materials and Systems

Paterna, Spain
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Maisanaba S.,University of Seville | Guzman-Guillen R.,University of Seville | Puerto M.,University of Seville | Gutierrez-Praena D.,University of Seville | And 2 more authors.
Journal of Hazardous Materials | Year: 2018

The clay montmorillonite (Mt) is among the nanofillers more frequently used for food packaging applications. The organomodification of clays with different modifiers, such as silanes, is an important step in the preparation of improved polymer/clay materials known as nanocomposites. However, the toxicological data about these nanofillers is still scarce. In the present study, an in vitro toxicological evaluation in Caco-2 cells of two silane-modified clays based on Mt, Clay3 and Clay4 (0–250 μg/ml), was performed. The cytotoxicity, cell death, genotoxicity and oxidative stress produced by both organoclays were evaluated after 24 and 48 h of exposure. Moreover, the migration extracts obtained from nanocomposites of polypropylene (PP) + Clay3 and only PP were also investigated. Only Clay4 induced cytotoxicity, showing a reduction of cell viability to 63% of the control, as well as oxidative stress in a concentration-dependent manner. Regarding the PP-Clay3 migration extract, no cytotoxic effects were observed after exposure to the tested concentrations (0–100%). Moreover, significant differences in the presence of Ca, Mg and Si compared to the PP extract were obtained, although migration levels were in accordance with the food contact materials regulation. These findings indicate that a case-by-case toxicological assessment of organoclays should be performed. © 2017 Elsevier B.V.


Maisanaba S.,University of Seville | Jorda-Beneyto M.,Area of Packaging Materials and Systems | Camean A.M.,University of Seville | Jos A.,University of Seville
Food and Chemical Toxicology | Year: 2016

Globally, food industries have made significant progress in order to increase the shelf-life of food products and have fewer economic losses. In this sense, the use of organomodified clays destined to be incorporated in polymer matrices play a novel role, leading to improved materials named nanocomposites with enhanced technological profiles. Due to the presence of these clays into the package, the safety of the consumers is a main concern. Cloisite®30B and Clay1 are two organomodified clays containing quaternary ammonium salts as modifiers, that can be potentially used to reinforce packaging polymers. Available toxicity data about these clays, specifically genotoxicity, is still limited and inconclusive in some aspects. Thus, the purpose of this work was to evaluate both clays ability to induce genomic instability through the cytokinesis block micronucleus cytome assay (CBMN) and for the first time, their influence in the modulation of several genes involved in genotoxicity and cell death mechanisms. Overall, no genotoxicity response was obtained in any case at the conditions tested. On the other hand, significant expression changes were observed on the genes selected. Nevertheless, further studies are highly needed to elucidate and increase the knowledge about the molecular mechanisms of clays toxicity. © 2015 Elsevier Ltd.


Llana-Ruiz-Cabello M.,University of Seville | Gutierrez-Praena D.,University of Seville | Pichardo S.,University of Seville | Moreno F.J.,University of Seville | And 3 more authors.
Food and Chemical Toxicology | Year: 2014

Essential oils used as additives in the food industry due to its flavour, antimicrobial and antioxidant properties. Therefore, human can be exposed orally to these compounds through the ingestion of foods. In this sense, the present work aims to assess toxicological effects of oregano essential oil on the digestive tract. In concrete, the cytotoxic effects of two components of the oregano essential oils, carvacrol and thymol, and their mixture, on the intestinal cells line Caco-2 after 24 and 48. h of exposure are studied. The basal cytotoxicity endpoints assayed (total protein content, neutral red uptake and the tetrazolium salt reduction) and the annexin/propidium iodide staining indicated that carvacrol and the mixture carvacrol/thymol induced toxic effects. Moreover, a morphological study was performed in order to determine the ultrastructural cellular damages caused by these substances. The main morphological alterations were vacuolated cytoplasm, altered organelles and finally cell death. In addition, although no cytotoxic effects were recorded for thymol at any concentration and time of exposure, ultrastructural changes evidenced cellular damage such as lipid degeneration, mitochondrial damage, nucleolar segregation and apoptosis. © 2013 Elsevier Ltd.


Maisanaba S.,University of Seville | Prieto A.I.,University of Seville | Pichardo S.,University of Seville | Jorda-Beneyto M.,Area of Packaging Materials and Systems | And 2 more authors.
Toxicology in Vitro | Year: 2015

Modern food packaging has made great advances as result of global trends and consumer preferences, which are oriented to obtain improved food quality and safety. In this regard, clay minerals, and mainly Montmorillonite (Mt) are attracting considerable interest in food packaging because of the improvements developed in mechanical and barrier properties. Hence, the present work aim to assess the toxicity of four Montmorillonite-based clay minerals, an unmodified clay, Cloisite®Na+ (CNa+), and three modified Mt clays: Cloisite®30B (C30B), a commercial clay, and Clay1 and Clay2, two novel modified organoclays developed by the Packaging, Transport, & Logistics Research Institute (ITENE). First, the cytotoxic effects were studied in the Human Umbilical Vein Endothelial Cells (HUVEC). In addition, the potential mutagenicity of the clays was evaluated by the Ames test. Clay1 did not induce any cytotoxic effects in HUVEC, although it exhibited potential mutagenicity in TA98 Salmonella typhimurium strain. In contrast, Clay2 produced cytotoxicity in endothelial cells but no mutagenicity was recorded. However, CNa+ was not cytotoxic neither mutagenic. And finally, C30B showed positive results in both assays. Therefore, results showed that clay minerals have a different toxicity profile and a case by case toxicity evaluation is required. © 2015 Elsevier Ltd.


Houtman J.,Wageningen University | Maisanaba S.,University of Seville | Puerto M.,University of Seville | Gutierrez-Praena D.,University of Seville | And 3 more authors.
Applied Clay Science | Year: 2014

Nowadays, the incorporation of organomodified clays based on montmorillonite into polymers intended for packaging industry is a reality. The final result is a polymer nanocomposite with enhanced barrier properties. Different organomodified clays are already commercially available and others new ones are being developed; however little is known about their safety.In the present work, the cytotoxic effects (a tetrazolium salt reduction and protein content) of three organomodified clays, Cloisite®20A, a commercial clay, and Clay 1 and Clay 2, two novel modified clays developed by the Packaging, Transport, & Logistics Research Institute, were evaluated in Caco-2 and HepG2 cells after 24 and 48. h of exposure. Our results showed that only Clay 2 induced toxic effects in both cell lines. The mean effective concentration was calculated for each case, showing Caco-2 to be more sensitive than HepG2. Moreover, in order to elucidate the toxicity mechanisms of Clay 2, different mechanistic biomarkers were investigated. Interleukin leakage and generation of intracellular reactive oxygen species were not observed, whereas glutathione content decreased in HepG2. DNA damage (comet assay) was induced in both cell lines at the highest concentration tested. Overall, results show that the type of clay, the concentrations range and the type of cell line play an important role in the toxicity observed. © 2014 Elsevier B.V.


Maisanaba S.,University of Seville | Hercog K.,Slovenian National Institute of Biology | Ortuno N.,Area of Packaging Materials and Systems | Jos A.,University of Seville | Zegura B.,Slovenian National Institute of Biology
Chemosphere | Year: 2016

Clay2 is an organomodified montmorillonite developed by the Technological Institute of Packaging, Transport and Logistic (ITENE) in order to improve polymeric materials used in food packaging. There is not much known on Clay2 toxic potential, particularly at DNA level, therefore it is mandatory to assess its toxicity prior to its commercialization. In the present study the human hepatoma cell line (HepG2) was exposed to non-cytotoxic concentrations of Clay2 and the genomic stability was studied with the Cytokinesis block micronucleus cytome assay, by determining the formation of micronuclei (MN), nucleoplasmic bridges (NPBs) and nuclear buds (NBUDs). Moreover, the expression of various genes involved in the mechanisms of its action using the real-time quantitative PCR was studied. The results obtained provide the evidence that Clay2 is potentially genotoxic as it increased the frequency of micronuclei. In addition it deregulated genes involved in the metabolism, immediate-early response/signaling, DNA damage and oxidative stress showing new valuable information on the cellular response to Clay2. Nonetheless, further studies are highly needed to elucidate the molecular mechanisms of clays toxicity. © 2016 Elsevier Ltd.


Llana-Ruiz-Cabello M.,University of Seville | Pichardo S.,University of Seville | Banos A.,DOMCA S.A. | Nunez C.,DOMCA S.A. | And 4 more authors.
LWT - Food Science and Technology | Year: 2015

New food packaging systems are been developed by food industry. The present work proposes the use of polylactic acid (PLA) containing different percentages of an Allium spp. extract (2%, 5%, and 6.5% Proallium ®) to be used in the packaging of ready-to-eat salads. Mechanical and optical properties of PLA films studied did not show remarkable changes when the active substance was incorporated. Additionally, no significant antioxidant activity was observed, although remarkable antimicrobial activity was recorded, mainly in films containing 5% and 6.5% of the Allium spp. extract. Moreover, the antimicrobial effect in vivo was observed in lettuce. All developed films decreased enterobacterial growth in a concentrationdependent manner. Regarding aerobic bacteria, film with the highest concentration of active agent (6.5%) was found to be effective up to 5 days of storage, and even 7 days for moulds. Therefore, the PLAProallium ® film could be a promising antimicrobial active material, especially for ready-to-eat salads. © 2015 Elsevier.


PubMed | Slovenian National Institute of Biology, University of Seville and Area of Packaging Materials and Systems
Type: | Journal: Chemosphere | Year: 2016

Clay2 is an organomodified montmorillonite developed by the Technological Institute of Packaging, Transport and Logistic (ITENE) in order to improve polymeric materials used in food packaging. There is not much known on Clay2 toxic potential, particularly at DNA level, therefore it is mandatory to assess its toxicity prior to its commercialization. In the present study the human hepatoma cell line (HepG2) was exposed to non-cytotoxic concentrations of Clay2 and the genomic stability was studied with the Cytokinesis block micronucleus cytome assay, by determining the formation of micronuclei (MN), nucleoplasmic bridges (NPBs) and nuclear buds (NBUDs). Moreover, the expression of various genes involved in the mechanisms of its action using the real-time quantitative PCR was studied. The results obtained provide the evidence that Clay2 is potentially genotoxic as it increased the frequency of micronuclei. In addition it deregulated genes involved in the metabolism, immediate-early response/signaling, DNA damage and oxidative stress showing new valuable information on the cellular response to Clay2. Nonetheless, further studies are highly needed to elucidate the molecular mechanisms of clays toxicity.


PubMed | IRNAS CSIC, University of Seville and Area of Packaging Materials and Systems
Type: Journal Article | Journal: Journal of the science of food and agriculture | Year: 2016

Environmental, economic and safety challenges motivate shift towards safer materials for food packaging. New bioactive packaging techniques, i.e. addition of essential plant oils (EOs), are gaining attention by creating barriers to protect products from spoilage. Analytical pyrolysis gas chromatography-mass spectrometry (Py-GC-MS) was used to fingerprint a bioactive polylactic acid (PLA) with polybutylene succinate (PBS) (950 g kg(-1) :50 g kg(-1) ) film extruded with variable quantities (0, 20, 50 and 100 g kg(-1) ) of Origanum vulgare EO.Main PLA:PBS pyrolysis products were lactide enantiomers and monomer units from the major PLA fraction and succinic acid anhydride from the PBS fraction. Oregano EO pyrolysis released cymene, terpinene and thymol/carvacrol peaks as diagnostic peaks for EO. In fact, linear correlation coefficients better than 0.950R(2) value (P < 0.001) were found between the chromatographic area of the diagnostic peaks and the amount of oregano EO in the bioplastic.The pyrolytic behaviour of a bio-based active package polymer including EO is studied in detail. Identified diagnostic compounds provide a tool to monitor the quantity of EO incorporated into the PLA:PBS polymeric matrix. Analytical pyrolysis is proposed as a rapid technique for the identification and quantification of additives within bio-based plastic matrices. 2015 Society of Chemical Industry.


PubMed | University of Seville and Area of Packaging Materials and Systems
Type: | Journal: Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association | Year: 2016

Globally, food industries have made significant progress in order to increase the shelf-life of food products and have fewer economic losses. In this sense, the use of organomodified clays destined to be incorporated in polymer matrices play a novel role, leading to improved materials named nanocomposites with enhanced technological profiles. Due to the presence of these clays into the package, the safety of the consumers is a main concern. Cloisite()30B and Clay1 are two organomodified clays containing quaternary ammonium salts as modifiers, that can be potentially used to reinforce packaging polymers. Available toxicity data about these clays, specifically genotoxicity, is still limited and inconclusive in some aspects. Thus, the purpose of this work was to evaluate both clays ability to induce genomic instability through the cytokinesis block micronucleus cytome assay (CBMN) and for the first time, their influence in the modulation of several genes involved in genotoxicity and cell death mechanisms. Overall, no genotoxicity response was obtained in any case at the conditions tested. On the other hand, significant expression changes were observed on the genes selected. Nevertheless, further studies are highly needed to elucidate and increase the knowledge about the molecular mechanisms of clays toxicity.

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