Potential application of grape (Vitis vinifera L.) stem extracts in the cosmetic and pharmaceutical industries: Valorization of a by-product
Introduction
Efficient use of resources through minimization of waste, long-term value retention, reduction of primary resources, and closed cycles of products within the limits of environmental protection and socioeconomic advantages can be defined as Circular Economy. This economic model enhances sustainable development while decreasing the negative consequences of lack of resources and environmental degradation (Morseletto, 2020). Various activities in the agricultural sector generate large amounts of by-products and wastes, where these wastes can cause sustainability problems due to the large quantities produced in a limited time period and due to the organic matter content (Coderoni and Perito, 2019). One of these activities is the grape production, reaching 73.3 million tons in 2017. It is estimated that 52 % of this production goes to the wine industry (OIV, 2018), where large quantities of by-products are generated, such as grape pomace, seeds, pulp, skins, leaves, stems, and wine lees (Gouvinhas et al., 2019). The recovery of these by-products/wastes has become increasingly important, providing multiple environmental, economic and social benefits, since their reuse generates the production of new added products, as a consequence of this circular approach (Coderoni and Perito, 2019).
Grape stem, which accounts for 25 % of total by-products, is the less characterized and valued of all by-products generated (Barros et al., 2014). This by-product is a rich source of phenolic compounds, celluloses, hemicelluloses and lignins (Gouvinhas et al., 2019). However, is it usually destined to the production of spirits, dietary fiber, vegetable protein concentrates, fertilizers (Domínguez-Perles et al., 2014), and animal feed (Sahpazidou et al., 2014).
In the last years, bioactive compounds present in by-products have attracted attention due to their health benefits, leading to a growing interest of the scientific community for the exploration of these residues (Gouvinhas et al., 2018). Phenolic compounds are secondary metabolites produced by plants that demonstrated to have several health benefits, acting as antioxidants, antimicrobials, anticarcinogens, antidiabetics, among others (Gouvinhas et al., 2019). Regarding to grape stems polyphenolic composition, several phenolic compounds were identified, such as flavonols (e.g. quercetin-3-O-rutinoside, kaempferol-3-O-glucoside), flavanols (e.g. catechin, epicatechin), anthocyanins (e.g. malvidin-3-O-glucoside, malvidin-3-O-(6-O-caffeoyl)-glucoside), phenolic acids (e.g. gallic, caffeic and caftaric acids), stilbenes (e. g. trans-resveratrol, Ɛ-viniferin) and procyanidins (e.g. procyanidin B2 and B3) (Anastasiadi et al., 2012; Barros et al., 2014; Dias et al., 2015; Domínguez-Perles et al., 2016; Gouvinhas et al., 2018; Sahpazidou et al., 2014).
The human body, when exposed to external conditions (for example pollutants, radiation and pathogens) produces a large quantity of reactive oxygen species (ROS), which causes oxidative stress, and may also induce inflammation (Zhang and Tsao, 2016). Phenolic compounds prevent or inhibit the production of ROS, suspending oxidation processes and repairing tissues deficiencies (Baenas et al., 2018). Moreover, while non-steroidal anti-inflammatory drugs are the most widely used in the world to fight inflammation, they have undesirable side-effects, for example on the kidneys and cardiovascular system, limiting their use. Thus, new studies are being performed to identify natural compounds that can be applied as anti-inflammatory agents or in the development of new drugs (Aouey et al., 2016). In addition, oxidative stress and inflammation are also related to aging processes (Martín-Ortega and Campos, 2019), where changes in the biosynthetic activity of skin cells, namely some constitutive and extracellular matrix proteins (e.g. collagen and elastin) arise, as well as in a simultaneous activity modulation of several aging-related enzymes (e.g. elastase, tyrosinase) (Aguilar-Toalá et al., 2019; Taghouti et al., 2018). The enzyme tyrosinase (also called monophenol monooxygenases), in humans, is responsible for melanin synthesis in melanocytes, but when overproduction of melanin occurs in the skin, pigmentation disorder, such as hyperpigmentation, over-tanning, age spots and melasma occur (Liyanaarachchi et al., 2018). Elastase is an enzyme that hydrolyses elastin, influencing the mechanical properties of connective tissues, thus when a high activity of this enzyme is observed, or less elastin is produced, the skin loses firmness and elasticity (Aguilar-Toalá et al., 2019).
Antibiotic resistance is a public health problem worldwide, having impact in morbidity and/or high mortality rates, especially in developing countries (Lima et al., 2019). Specific microorganisms such as Listeria monocytogenes and Escherichia coli are present frequently in processed food, constituting a health risk (Gutiérrez-del-Río et al., 2018). In addition, one of the main causes of hospitalization worldwide are diabetic foot ulcer infections, mostly caused by Staphylococcus aureus (Zenão et al., 2017). To combat bacterial resistance, alternative antibiotics should be searched and developed to overcome the effectiveness of those currently available (Lima et al., 2019).
Thus, in order to replace synthetic drugs/antibiotics with more effective natural compounds, there is a growing demand to find natural compounds that can be used in the development of new products. Grapes stem may be integrated into this demand due to its polyphenolic content, thus enhancing sustainable development, with advantages for both the environment and the corporate economy. In this way, the aim of this study was to evaluate the scavenging activity and the antimicrobial activity of grape stem extracts, against gastrointestinal and diabetic foot wound bacteria. Furthermore, the anti-inflammatory and the anti-aging activities, allowing to prove if this by-product will be a good bet to produce new formulations. This investigation allowed to determine, for the first time, not only anti-tyrosinase and anti-elastase activities of grape stems, but also their capacity to inhibit the growth of foot wound ulcers bacteria.
Section snippets
Chemicals
The compounds 2,2-diphenyl-1-picrylhidrazyl radical (DPPH), 2,2-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid)diammonium salt (ABTS+), 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox), potassium persulfate, 2,4,6-Tris(2-pyridyl)-s-triazine (TPTZ), ferric chloride, and the enzymes tyrosinase, elastase, and reagents for all enzymatic activities were purchased from Sigma-Aldrich (Steinheim, Germany). Saline water (0.9 % NaCl), methanol, and sulfanilamide were acquired from Merck
Phenolic compounds identified and quantified in grape stem extracts
The identification of phenolic compounds by RP-HPLC-DAD was realized by comparison with pure standards of DAD spectra, retention times, and literature data (Anastasiadi et al., 2009; Gouvinhas et al., 2018; Queiroz et al., 2017). In grape stem extracts, eleven phenolic compounds were identified and quantified, belonging to different classes, namely hydroxybenzoic and hydroxycinnamic acids, flavanols, flavonols, stilbenes, and anthocyanins (Table 1). When analyzing the concentrations obtained,
Conclusions
The biological potential of grape stems was proven, once again, in this study. This by-product showed antioxidant activity, revealing a good scavenging capacity; antimicrobial activity, which showed high efficacy against Gram-positive bacteria, especially S. aureus and E. faecalis; anti-inflammatory activity, in which all extracts were shown to inhibit the NO production at non-toxic cellular concentrations. Finally, anti-aging activity was also found, where important results were obtained for
CRediT authorship contribution statement
Carla Leal: Methodology, Software, Investigation, Writing - original draft. Irene Gouvinhas: Conceptualization, Methodology, Investigation, Funding acquisition, Writing - review & editing, Supervision. Rafaela A. Santos: Investigation, Methodology. Eduardo Rosa: Funding acquisition. Amélia M. Silva: Writing - review & editing, Supervision. Maria José Saavedra: Supervision. Ana I.R.N.A. Barros: Conceptualization, Funding acquisition, Supervision.
Declaration of Competing Interest
None.
Acknowledgements
This work was supported by National Funds by FCT - Portuguese Foundation for Science and Technology, under the project UIDB/04033/2020 and the I&D project Interact - Integrative Research in Environment, Agro-Chains and Technology (NORTE-01-0145-FEDER-000017), regarding the research line “Fostering viticulture sustainability for Douro Valley: multidisciplinary efforts from field to wine (VitalityWINE)”, and the research line “Innovation for Sustainable Agro-food Chains (ISAC)” co-founded by the
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