Browsing by Author "Pinto, Diana"
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- Appraisal of a new potential antioxidants-rich nutraceutical ingredient from chestnut shells through in-vivo assays – A targeted metabolomic approach in phenolic compoundsPublication . Pinto, Diana; Almeida, Andreia; López-Yerena, Anallely; Pinto, Soraia; Sarmento, Bruno; Lamuela-Raventós, Rosa; Vallverdú-Queralt, Anna; Delerue-Matos, Cristina; Rodrigues, FranciscaChestnut (Castanea sativa) shells (CSS) are a source of bioactive compounds with well demonstrated in-vitro antioxidant properties. Nevertheless, no in-vivo studies have already evaluated this effect. This study evaluated the effects of the oral daily administration of an eco-friendly CSS extract (50 and 100 mg/kg per body weight (b. w.)) to rats regarding in-vivo antioxidant activity, glucose and lipids levels, and metabolomic profiling of poly- phenols by LC-ESI-LTQ-Orbitrap-MS. The results demonstrated the in-vivo antioxidant properties in the animals liver, kidney and blood serum, as well as protective effects against hemolysis and rising of blood glucose and lipids levels. New insights on metabolomic profiling of polyphenols proved their absorption and further biotransformation by phase I (hydrogenation and hydroxylation) and II reactions (glucuronidation, methylation and sulfation). This is the first study that attempted to validate a novel nutraceutical ingredient extracted from CSS by in-vivo assays, corroborating the outcomes screened by in-vitro assays
- Castanea sativa shells: A review on phytochemical composition, bioactivity and waste management approaches for industrial valorizationPublication . Pinto, Diana; Cádiz-Gurrea, María de la Luz; Vallverdú-Queralt, Anna; Delerue-Matos, Cristina; Rodrigues, FranciscaCastanea sativa is an outstanding species that represents a valuable natural resource for rural populations. C. sativa shells (CSS), an abundant agro-industrial by-product generated during chestnut peeling process, is commonly discarded or used as fuel. Nevertheless, CSS produced are not depleted by this application and huge amounts are still available, being particularly rich in bioactive compounds (polyphenols, vitamin E, lignin and oligosaccharides) with health benefits. Phytochemical studies reported not only antioxidant and antimicrobial activities, but also anti-inflammatory, anticancer, hypolipidemic, hypoglycemic and neuroprotective activities. The application of a suitable extraction technique is required for the isolation of bioactive compounds, being green extraction technologies outstanding for the industrial recovery of chestnut shells' bioactive compounds. CSS were highlighted as remarkable sources of functional ingredients with promising applications in food and nutraceutical fields, mainly as natural antioxidants and effective prebiotics. This review aims to summarize the phytochemical composition and pro-healthy properties of CSS, emphasizing the sustainable extraction techniques employed in the recovery of bioactive compounds and their potential applications in food and nutraceutical industries.
- Chemical characterization and bioactive properties of a coffee-like beverage prepared from Quercus cerris kernelsPublication . Pinto, Diana; Diaz Franco, Santiago; Silva, Ana Margarida; Cupara, Snezana; Koskovac, Marijana; Kojicic, Ksenija; Soares, Sónia; Rodrigues, Francisca; Sut, Stefania; Dall'Acqua, Stefano; Oliveira, M Beatriz P PIn the present study, a coffee-like beverage was prepared from Quercus cerris seeds. Seeds presented 60.4% carbohydrates (of which 26.9% is fiber), 6.3% proteins and 4.0% fat. In the coffee-like beverage, the TPC and TFC were 2070.2 mg GAE per L and 285.3 mg CEQ per L, respectively. The IC50 values determined using FRAP and DPPH assays were, respectively, 203.1 mg L-1 and 271.6 mg L-1. Concerning the reactive species scavenging capacity, the extract was more effective against O2˙-, HClO and ˙NO (namely, IC50 = 17.24 ± 0.24 μg mL-1, IC50 = 9.25 ± 0.92 μg mL-1 and IC50 = 0.65 ± 0.06 μg mL-1). Qualitative and quantitative analyses by 1D- and 2D-NMR and LC-MS indicated high levels of ellagic acid and gallotannin or ellagitannin derivatives. The optimal range of non-cytotoxic concentrations in Caco-2 and HT29-MTX cell lines was between 0.1 and 1.0 μg mL-1. This is the first study that evaluates the Q. cerris seed extract as a beverage.
- Development and Characterization of Functional Cookies Enriched with Chestnut Shells Extract as Source of Bioactive Phenolic CompoundsPublication . Pinto, Diana; Moreira, Manuela M.; Vieira, Elsa F.; Švarc-Gajić, Jaroslava; Vallverdú-Queralt, Anna; Brezo-Borjan, Tanja; Delerue-Matos, Cristina; Rodrigues, FranciscaChestnut (Castanea sativa) shells (CSs), an undervalued agro-industrial biowaste, have arisen as a source of bioactive compounds with promising health-promoting effects. This study attempted, for the first time, to develop a functional food, namely cookies, using a CS extract obtained by an eco-friendly technology (subcritical water extraction). The cookies were characterized regarding their nutritional composition, total phenolic and flavonoid contents (TPC and TFC, respectively), antioxidant/antiradical activities, phenolic profile, and sensory evaluation. The results demonstrated that the CS-extract-enriched cookies were mainly composed of carbohydrates (53.92% on dry weight (dw)), fat (32.62% dw), and fiber (5.15% dw). The phenolic profile outlined by HPLC-PDA revealed the presence of phenolic acids, flavonoids, and hydrolysable tannins, attesting to the high TPC and TFC. The in vitro antioxidant/antiradical effects proved the bioactivity of the functional cookies, while the sensory evaluation unveiled excellent scores on all attributes (≥6.25). The heatmap diagram corroborated strong correlations between the TPC and antioxidant/antiradical properties, predicting that the appreciated sensory attributes were closely correlated with high carbohydrates and phenolic compounds. This study encourages the sustainable recovery of antioxidants from CSs and their further employment as an active nutraceutical ingredient in functional cookies.
- Evaluation of the Extraction Temperature Influence on Polyphenolic Profiles of Vine-Canes (Vitis vinifera) Subcritical Water ExtractsPublication . Dorosh, Olena; M. Moreira, Manuela; Pinto, Diana; F. Peixoto, Andreia; Freire, Cristina; Costa, Paulo; Rodrigues, Francisca; Delerue-Matos, CristinaThis work focused on evaluating the possibility of using vineyard pruning wastes from two Portuguese Vitis vinifera varieties; Touriga Nacional (TN) and Tinta Roriz (TR), as new potential ingredients for the nutraceutical industry. An environmentally friendly extraction technique; namely subcritical-water extraction (SWE), was employed. The overall results indicate that phenolic acids were the major class of compounds quantified; being gallic acid the principal one. The highest value for total phenolic content (TPC) was obtained for the TR extract at 250 °C (181 ± 12 mg GAE/g dw). In terms of antioxidant activity; the DPPH values for the extracts obtained at 250 °C were approximately 4-fold higher than the ones obtained at 125 °C; with TR extract presenting the highest value (203 ± 22 mg TE/g dw). Thus, the TR extract obtained through SWE at 250 °C was selected to evaluate the scavenging activity and the in vitro effects on cells due to the best results achieved in the previous assays. This extract presented the ability to scavenge reactive oxygen species (O2●-, HOCl and ROO●). No adverse effects were observed in HFF-1 viability after exposure to extract concentrations below 100 μg/mL. This work demonstrated that vine-canes extracts could be a potential ingredient to nutraceutical industry
- From soil to cosmetic industry: Validation of a new cosmetic ingredient extracted from chestnut shellsPublication . Pinto, Diana; de la Luz Cádiz-Gurrea, María; Garcia, Juliana; Saavedra, Maria José; Freitas, Vitor; Costa, Paulo; Sarmento, Bruno; Delerue-Matos, Cristina; Rodrigues, FranciscaSustainability arises as a challenge in the last decades and the cosmetic field is not an exception. The search for new cosmetic active principles extracted from food byproducts through eco-innovative technologies has emerged as a novel concept. However, the European Regulation is very restrictive and different assays should be performed to guarantee the products' efficacy and safety. Considering a multidisciplinary approach, this paper aims to compare chestnut shell extracts prepared by three different green techniques – Subcritical Water Extraction (SWE), Ultrasound-Assisted Extraction (UAE) and Supercritical Fluids Extraction (SFE) – regarding antimicrobial activity, hyaluronidase and elastase inhibitory activities, in-vitro cytotoxicity on dermal cells (HaCaT and HFF-1) and skin permeation (using Franz cells coupled to human skin). The best extract was tested on skin and ocular 3D models and through an in-vivo patch test to evaluate its irritancy and safety. The results demonstrated the antimicrobial properties of the extracts (particularly against Staphylococcus aureus) as well as the capacity to inhibit the hyaluronidase (IC50 = 0.76–54.36 mg/mL) and elastase (33.56–82.70% at 0.4 mg/mL) activities. Regarding in-vitro cell assays, UAE and SWE extracts revealed the best outcomes. Between 0.1 and 100 μg/mL, none of the extracts conducted to a decrease of cell viability in both cell lines (HaCaT and HFF-1). The ex-vivo assay proved that SWE extract led to a higher polyphenol's permeation (1061.6 μg/g dw), being selected for further assays. Ellagic acid was the major polyphenol that permeated the human skin (732.1–847.0 μg/g dw). The 3D skin and ocular models evidenced absence of irritation by SWE extract, without decreasing cell viability and releasing IL-1α. Finally, an in-vivo patch test confirmed the absence of irritation in humans based on the acute irritation index of 0.50 achieved 24 h after the patch removal. To the best of our knowledge, this is the first study that validated by in-vitro and in-vivo assays, and in accordance with the European Regulation 1223/2009, a new cosmetic ingredient extracted from chestnut shells.
- Green-Sustainable Recovery of Phenolic and Antioxidant Compounds from Industrial Chestnut Shells Using Ultrasound-Assisted Extraction: Optimization and Evaluation of Biological Activities In VitroPublication . Lameirão, Fátima; Pinto, Diana; F. Vieira, Elsa; F. Peixoto, Andreia; Freire, Cristina; Sut, Stefania; Dall’Acqua, Stefano; Costa, Paulo; Delerue-Matos, Cristina; Rodrigues, FranciscaChestnut processing industry generates large amounts of by-products, including leaves, burs and shells that are a source of bioactive compounds. The purpose of this study was to establish an ultrasound-assisted extraction (UAE) of phenolic and antioxidant compounds from industrial chestnut shells. A central composite design (CCD) was conducted to analyze the effects of time (4–46 min) and temperature (34–76 °C) in the antioxidant activity (2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS), 2,2-diphenyl-1-picrylhydrazyl (DPPH), and ferric reducing antioxidant power (FRAP)) and total phenolic compounds (TPC) of chestnut shells extracts. The optimal extraction conditions were obtained at 70 °C for 40 min. The optimal extract was characterized regarding phenolic profile, radical scavenging capacity, and effects on intestinal and dermal cell lines. The optimal extract revealed high amounts of ellagic acid (40.4 µg/mg dw), followed by caffeic acid derivative (15.4 µg/mg dw) and epigallocatechin (15.3 µg/mg dw). Indeed, the extract exhibited the highest scavenging efficiencies against NO● (IC50 = 0.1 µg/mL) and HOCl (IC50 = 0.7 µg/mL) and did not conducted to a decrease on HaCaT and HFF-1 viability up to 100 μg/mL. Oppositely, a decrease on Caco-2 and HT29-MTX viability was observed. This study suggests that UAE could be a sustainable option to valorize chestnut shells as raw material for different industries.
- Infusions and decoctions of dehydrated fruits of Actinidia arguta and Actinidia deliciosa: Bioactivity, radical scavenging activity and effects on cells viabilityPublication . Silva, Ana Margarida; Pinto, Diana; Fernandes, Iva; Albuquerque, Tânia Gonçalves; Costa, Helena S.; Freitas, Vitor; Rodrigues, Francisca; Oliveira, M. Beatriz P.P.Actinidia deliciosa and A. arguta fruits (kiwifruit and kiwiberry, respectively) are an excellent source of bioactive compounds. The aim of this paper is to valorize the fruits that are not commercialized (e.g. due to inadequate size or physical damage) in infusions and decoctions. The antioxidant activity, the scavenging activity against reactive species, the phenolic profile and the intestinal effects of infusions and decoctions of dehydrated fruits were evaluated and compared. Decoctions presented the highest antioxidant activity and a good ability to capture HOCl and NO. The phenolic composition of A. arguta present quinic acid, cis-caftaric acid and its derivatives, caffeoyl hexoside, luteolin glucuronide, quercetin derivatives and myristin, while A. deliciosa extracts were characterized by the presence of quinic acid, caffeic acid and its derivatives and caffeoyl hexoside. No adverse effects were observed on Caco-2 and HT29-MTX cells. Kiwiberry decoctions showed to be the best option to keep the fruits benefits.
- New Perspectives on the Sustainable Employment of Chestnut Shells as Active Ingredient against Oral Mucositis: A First ScreeningPublication . Ferreira, Ana Sofia; Silva, Ana Margarida; Pinto, Diana; Moreira, Manuela M.; Ferraz, Ricardo; Švarc-Gajić, Jaroslava; Costa, Paulo C.; Delerue-Matos, Cristina; Rodrigues, FranciscaOral mucositis (OM), a common side effect of oncological treatment, is an oral mucosal disorder characterized by painful ulcerations and increased risk of infection. The use of natural antioxidants to suppress the redox imbalance responsible for the OM condition has emerged as an interesting approach to prevent/treat OM. This study aims to explore the chestnut (Castana sativa) shells as potential active ingredient against OM. Therefore, chestnut shells were extracted at different temperatures (110–180 °C) by Subcritical Water Extraction (SWE), aiming to recover antioxidants. The extracts were also evaluated against microorganisms present in the oral cavity as well as on human oral cell lines (TR146 and HSC3). The highest phenolic content was obtained with the extraction temperature of 110 °C, exhibiting the best antioxidant/antiradical activities and scavenging efficiencies against HOCl (IC50 = 4.47 μg/mL) and ROO• (0.73 μmol TE/mg DW). High concentrations of phenolic acids (e.g., gallic and protocatechuic acids) and flavanoids (catechin, epicatechin and rutin) characterized the phenolic profile. The antimicrobial activity against several oral microorganisms present in the oral cavity during OM, such as Streptococcus, Staphylococcus, Enterococcus, and Escherichia, was demonstrated. Finally, the effects on HSC3 and TR146 cell lines revealed that the extract prepared at 110 °C had the lowest IC50 (1325.03 and 468.15 µg/mL, respectively). This study highlights the potential effects of chestnut shells on OM
- Potential therapeutic application of chestnut shells in oral mucositis treatment: Green extraction of polyphenolsPublication . Ferreira, Ana Sofia; Silva, Ana Margarida; Pinto, Diana; Ferraz, Ricardo; Moreira, Manuela M.; Švarc-Gajić, Jaroslava; Costa, Paulo; Delerue-Matos, Cristina; Rodrigues, FranciscaOral mucositis (OM), a common side effect of oncological treatment, is an inflammatory condition characterized by painful ulcerations, difficulty swallowing and speaking, and increased risk of infection, which can compromise the primary treatment outcome. The use of natural antioxidants to inhibit theredox imbalance responsible for OM development has emerged as an interesting approach for OM prevention and treatment. This study aims to explore chestnut (Castana sativa) shells (by-products) aspotential active ingredient against OM [2]. To recover antioxidants using sustainable technology, chestnut shells were extracted using Subcritical Water Extraction (SWE) at different temperatures (110-180 °C). The extracts were evaluated against microorganisms present in the oral cavity as well as in human oral cell lines (TR146 and HSC3). The optimal extraction temperature was 110 °C, exhibiting thehighest phenolic content and the best antioxidant/antiradical activities and scavenging efficiencies against HOCl (IC50 = 4.47 μg/mL) and ROO● (0.73 μmol TE/mg DW). High concentrations of phenolicacids (e.g., gallic and protocatechuic acid) and flavanoids (catechin, epicatechin and rutin) characterizedthe phenolic profile. Antimicrobial activity against several microorganisms present in the oral cavity during OM, such as Streptococcus, Staphylococcus, Enterococcus, and Escherichia, was also demonstrated. The effects on HSC3 and TR146 cell lines revealed that the extract prepared at 110 °C showed the lowest IC50 (1325.03 and 468.15 µg/mL, respectively). Therefore, this extract was selectedfor encapsulation and incorporation into an oral film as buccal drug delivery system. Methocel 1000 demonstrated ideal physicochemical properties from different set of polymers, such as thickness, mass, content uniformity, dissolution time in artificial saliva, superficial pH, folding, tensile strength, and elongation.