Peroxidases are widespread key antioxidant enzymes that catalyse the oxidation of electron donor substrates in parallel with the decomposition of H 2 O 2 . In this work, a novel tomato peroxidase, named SAAP2, was isolated from MicroTom cell cultures, purified, and characterised. The enzyme was identified with 64% sequence coverage as the leprx21 gene product (suberization-associated anionic peroxidase 2-like) from Solanum lycopersicum , 334 amino acids long. Compared to other plant peroxidases, SAAP2 was more active at elevated temperatures, with the optimal temperature and pH at 90 °C and 5.0, respectively. Furthermore, the enzyme retained more than 80% of its maximal activity over the range of 70–80 °C and the presence of NaCl (1.0–4.5 M). It also exhibited broad pH versatility (65% relative activity over the pH range 2.0–7.0), acid-tolerance (80% residual activity after 22 h at pH 2.0–7.0), high thermostability (50% residual activity after 2 h at 80 °C) and proteolytic resistance. SAAP2 exhibited exceptional resistance under thermo-acidic conditions compared to the horseradish peroxidase benchmark, suggesting that it may find potential applications as a supplement or anti-pollution agent in the food industry.

Superoxide dismutase (SOD) is a fundamental antioxidant enzyme that neutralises superoxide ions, one of the main reactive oxygen species (ROS). Extremophile organisms possess enzymes that offer high stability and catalytic performances under a wide range of conditions, thus representing an exceptional source of biocatalysts useful for industrial processes. In this study, SODs from the thermo-halophilic Aeropyrum pernix (SODAp) and the thermo-acidophilic Saccharolobus solfataricus (SODSs) were heterologously expressed in transgenic tomato cell cultures. Cell extracts enriched with SODAp and SODSs showed a remarkable resistance to salt and low pHs, respectively, together with optimal activity at high temperatures. Moreover, the treatment of tuna fillets with SODAp-extracts induced an extension of the shelf-life of this product without resorting to the use of illicit substances. The results suggested that the recombinant plant extracts enriched with the extremozymes could find potential applications as dietary supplements in the nutrition sector or as additives in the food preservation area, representing a more natural and appealing alternative to chemical preservatives for the market.

Psychological stress activates catecholamine production, determines oxidation processes, and alters the lipid barrier functions in the skin. Scientific evidence associated with the detoxifying effect of fruits and vegetables, the growing awareness of the long-term issues related to the use of chemical-filled cosmetics, the aging of the population, and the increase in living standards are the factors responsible for the growth of food-derived ingredients in the cosmetics market. A Ficus carica cell suspension culture extract (FcHEx) was tested in vitro (on keratinocytes cells) and in vivo to evaluate its ability to manage the stress-hormone-induced damage in skin. The FcHEx reduced the epinephrine (−43% and −24% at the concentrations of 0.002% and 0.006%, respectively), interleukin 6 (−38% and −36% at the concentrations of 0.002% and 0.006%, respectively), lipid peroxide (−25%), and protein carbonylation (−50%) productions; FcHEx also induced ceramide synthesis (+150%) and ameliorated the lipid barrier performance. The in vivo experiments confirmed the in vitro test results. Transepidermal water loss (TEWL; −12.2%), sebum flow (−46.6% after two weeks and −73.8% after four weeks; on the forehead −56.4% after two weeks and −80.1% after four weeks), and skin lightness (+1.9% after two weeks and +2.7% after four weeks) defined the extract’s effects on the skin barrier. The extract of the Ficus carica cell suspension cultures reduced the transepidermal water loss, the sebum production, the desquamation, and facial skin turning to a pale color from acute stress, suggesting its role as an ingredient to fight the signs of psychological stress in the skin.

Skin aging is a very well-known process setting a gradual worsening of skin mechanical features due to a decline in the production of the extra-cellular matrix machinery and to a concurrent change in the contraction process. To slow this progression, it is crucial to induce the expression of several proteins able to promote elastic fibers formation and tissue repair. Here, the Oenothera biennis cell culture aqueous extract has been investigated from a chemical point of view and then it was tested in vitro, in cell, and in ex vivo experiments as adjuvant in counteracting skin aging. Accordingly, it has been shown that the Oenothera biennis extract was able, by increasing MYLK gene expression, to promote matrix collagen contraction, actin polymerization, and the production of essential ECM proteins.

Facial pore enlargement is considered a significant esthetic and health concern in skincare cosmetics. The pores fulfill the critical function of keeping the skin surface hydrated and protected against microbial infections. The hyperseborrhea, the stress factors, and the hormonal triggers can cause pore size enlargement, causing higher susceptibility of the skin to microbe aggressions and inflammatory reactions. Thus, reducing excessive sebum production and keeping functional pores are two of the most requested activities in skincare cosmetics. A Cirsium eriophorum cell culture extract was investigated for its role in sebum regulation, stratum corneum desquamation, and anti‐inflammation. The extract was able to regulate essential markers associated with sebum secretion and pore enlargements, such as the enzyme 5α‐reductase, which plays a central role in sebum production, and the trypsin‐like serine protease Kallikrein 5, which promotes skin exfoliation and antimicrobial response. Moreover, the extract showed a sebum‐normalizing and pore refining activity in individuals having seborrheic or acne‐prone skins, suggesting a role of the C. eriophorum extract in rebalancing altered skin conditions responsible for pore enlargement.

Plant extracts are rich in bioactive compounds, such as polyphenols, sesquiterpenes, and triterpenes, which potentially have antiviral activities. As a consequence of the coronavirus disease 2019 pandemic, caused by the severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) virus, thousands of scientists have been working tirelessly trying to understand the biology of this new virus and the disease pathophysiology, with the main goal of discovering effective preventive treatments and therapeutic agents. Plant-derived secondary metabolites may play key roles in preventing and counteracting the rapid spread of SARS-CoV-2 infections by inhibiting the activity of several viral proteins, in particular those involved in the virus entry into the host cells and its replication. Using in vitro approaches, we investigated the role of a pomegranate peel extract (PPE) in attenuating the interaction between the SARS-CoV-2 Spike glycoprotein and the human angiotensin-converting enzyme 2 receptor, and on the activity of the virus 3CL protease. Although further studies will be determinant to assess the efficacy of this extract in vivo, our results opened new promising opportunities to employ natural extracts for the development of effective and innovative therapies in the fight against SARS-CoV-2.

Annalisa Tito 1 Massimiliano Minale Salvatore Riccio 2 Fabrizia Grieco 2 Maria Gabriella Colucci Fabio Apone 1Arterra Bioscience Srl, Naples, 80142, Italy; 2Farmaceutici Damor S.p.A., Naples 80145, Italy; 3VitaLab Srl, Naples, 80142, Italy Introduction: Chronic skin lesions represent a problem of increasing occurrence, mostly due to the global ageing of the world population. Research in skin care and dermatology is constantly looking for new non-invasive solutions, preferably those based on the use of natural certified products, able to accelerate the spontaneous skin repair mechanisms and without altering the skin normal appearance and functionality. The wound healing process in the skin is finely regulated by several factors and orchestrated mechanisms, which modulate the progression and the fitting of different consequent phases, including haemostasis, inflammation, cell proliferation and tissue remodelling. It was previously shown that a patented Triticum vulgare aqueous extract was able to trigger the skin repair process by stimulating new tissue growth and reducing the expression levels of inflammatory mediators, such as IL6, TNFα, prostaglandin E2, and nitric oxide. Methods: Scratch assay was performed in Human Dermal Fibroblasts (HDF). The production of fibronectin was measured by gene expression, protein quantification and localization using specific antibodies in HDF. The polymerization of actin was measured using rhodaminphalloidin in HDF. The epidermal lipid content was estimated in HaCaT (human spontaneously immortalized keratinocytes) using Nile Red staining and the increasing GBA gene expression and activity was demonstrated by RT-PCR and enzymatic activity assay. Results: In the present study, it was demonstrated that the T. vulgare extract enhanced cell migration inducing the synthesis of fibronectin, new actin polymerization and stimulating the expression of the Hyaluronan Synthase 2. Furthermore it improved the restoration of the epidermal barrier stimulating lipid synthesis. Conclusion: In conclusion, we demonstrated that the T. vulgare extract possessed promising potential to be developed as a wound healing promoting agent in skin care and dermatology.

Introduction: Chronic skin lesions represent a problem of increasing occurrence, mostly due to the global ageing of the world population. Research in skin care and dermatology is constantly looking for new non-invasive solutions, preferably those based on the use of natural certified products, able to accelerate the spontaneous skin repair mechanisms and without altering the skin normal appearance and functionality. The wound healing process in the skin is finely regulated by several factors and orchestrated mechanisms, which modulate the progression and the fitting of different consequent phases, including haemostasis, inflammation, cell proliferation and tissue remodelling. It was previously shown that a patented Triticum vulgare aqueous extract was able to trigger the skin repair process by stimulating new tissue growth and reducing the expression levels of inflammatory mediators, such as IL-6, TNF alpha, prostaglandin E2, and nitric oxide. Methods: Scratch assay was performed in Human Dermal Fibroblasts (HDF). The production of fibronectin was measured by gene expression, protein quantification and localization using specific antibodies in HDF. The polymerization of actin was measured using rhodaminphalloidin in HDF. The epidermal lipid content was estimated in HaCaT (human spontaneously immortalized keratinocytes) using Nile Red staining and the increasing GBA gene expression and activity was demonstrated by RT-PCR and enzymatic activity assay. Results: In the present study, it was demonstrated that the T. vulgare extract enhanced cell migration inducing the synthesis of fibronectin, new actin polymerization and stimulating the expression of the Hyaluronan Synthase 2. Furthermore it improved the restoration of the epidermal barrier stimulating lipid synthesis. Conclusion: In conclusion, we demonstrated that the T. vulgare extract possessed promising potential to be developed as a wound healing promoting agent in skin care and dermatology.

Grapes (Vitis vinifera) are one of the most widely-grown fruit crops in the world. Approximately 75% of the whole grape production is utilized into winemaking, while 25% of the original grape weight is grape pomace (skin, seeds and stalks), which is a by-product. The oil-soluble fraction of grape pomace is rich in phenolic compounds including flavonoids, carotenoids, phenolic acids, tannins, and stilbenes, high unsaturated fatty acids, vitamins (E, A and C) and phytosterols with economic importance to cosmetic, food and pharmaceutical industry. The objective of this study was to evaluate the cosmetic efficacy on skin balance, normalization, hydration and elasticity and on wrinkles prevention of dermocosmetic formulations containing grape seed oil extract on young and mature skin using in vitro and in vivo techniques.In vitro studies showed anti-aging effect through enhancement of cell detoxification, hydration, protection of cellular proteins, stabilization of connective tissues from UV-stress inducing hyaluronic acid synthase and stimulating collagen I and III synthesis.In vivo studies displayed wrinkles volume and depth reduction and a barrier protective effect inducing a high hydration of the skin.

In order to support their sessile lifestyle, plants evolved sophisticated defence mechanisms that result in the synthesis of a vast spectrum of chemical compounds. Plants have been the main source of natural ingredients for human use since ancient times and nowadays, with the development of plant tissue culture techniques, remarkable progresses towards the production of bioactive compounds on an industrial scale have been achieved, as plant tissue cultures represent standardized, contaminant-free, and bio-sustainable systems. In cosmetics, particularly as skin care applications, plant tissue cultures represent a valuable and promising source of active ingredients with a wide range of biological activities. While plant cells, grown as undifferentiated suspension cultures, could be a valuable source of active molecules which are normally synthesized in low amounts in differentiated tissues, hairy roots or somatic embryo cultures, due to their more specialized nature, synthetize compounds that can address more specific functions in skin care; although widespread in the pharmaceutical industry, plant tissue culture technology is not yet very common in the cosmetic field. The aim of the present article is to focus on the successful research accomplishments in the development of plant tissue cultures-derived active ingredients for cosmetic applications.

An overproduction of free radicals or reactive oxygen species, often due to environmental factors, can alter the DNA structure and irreversibly modify proteins and lipids in the living cells. The superoxide anion (O2-) is one of the strongest oxidant molecules produced under oxidative stress conditions but it can be neutralized by the action of the enzymes SuperOxide Dismutases (SODs). In all the human tissues, SODs are essential for the prevention of serious diseases and the protection against oxidative stress damages. In the dermo-cosmetic sector, SODs have found promising applications, but their use is limited due to the loss of activity following the addition of the enzyme in the skin care formulas and the exposure of the skin to UV radiations and heat. Extremophile organisms, which proliferate in extreme physical and/or geochemical conditions, represent a potential source of stable SOD enzymes, able to function even in harsh conditions of high temperature, acid pH and long UV exposures. In the present study we investigated on a Mn-SOD deriving from the extremophilic bacterium Deinococcus radiodurans and, after its expression in E.coli, the Mn-SOD was characterized in terms of chemical and physical properties. Its extraordinary features in terms of UV resistance prompted us to investigate further about its potential applications in the dermo-cosmetic sector. It was expressed in Solanum lycopersicum (tomato) cell cultures with the main goal of developing a new ingredient, capable of keeping its ROS neutralizing activity once exposed to UV radiations and even when added to skin care formulas.

Bioactive peptides (BP) are specific protein fragments that are physiologically important for most living organisms. It is proven that in humans they are involved in a wide range of therapeutic activities as antihypertensive, antioxidant, anti-tumoral, anti-proliferative, hypocholesterolemic, and anti-inflammatory. In plants, BP are involved in the defense response, as well as in the cellular signaling and the development regulation. Most of the peptides used as ingredients in health-promoting foods, dietary supplements, pharmaceutical, and cosmeceutical preparations are obtained by chemical synthesis or by partial digestion of animal proteins. This makes them not fully accepted by the consumers because of the risks associated with solvent contamination or the use of animal derived substances. On the other hand, plant and microalgae derived peptides are known to be selective, effective, safe, and well tolerated once consumed, thus they have got a great potential for use in functional foods, drugs, and cosmetic products. In fact, the interest in the plant and microalgae derived BP is rapidly increasing and in this review, we highlight and discuss the current knowledge about their studies and applications in the cosmetic field.

Ageing is a complex and progressive phenomenon, during which the accumulation of morphological and chemical changes seriously compromises the capacity of the cells to proliferate and fulfil their biological tasks. The increase in the average age of the world population, associated with a higher occurrence of age-related diseases, is prompting scientific research to look for new strategies and molecular targets that may help in alleviating age-related phenotypes. Growth factors, responsible for modulating several aging markers in many tissues and organs, represent valuable targets to fight age-associated dysfunctions. The growth differentiation factor GDF11, a TGF-β family member, has been associated with the maintenance of youth phenotypes in different human tissues and organs, and in the skin has been related to an inhibition of the inflammatory response. We investigated the role of GDF11 in skin dermal fibroblasts, and we observed that its expression and activity were reduced in fibroblasts deriving from adult donors compared to neonatal ones. The main effect of GDF11 was the induction of collagen I and III, in both neonatal and adult fibroblasts, by triggering Smad signalling in a TGF-β-like fashion. Moreover, by analysing a number of plant extracts having GDF11 inducing activity, we found that a peptide/sugar preparation, obtained from Lotus japonicus somatic embryo cultures, was capable of restoring GDF11 expression in older fibroblasts and to activate the synthesis of collagen I, collagen III and periostin, an important protein involved in collagen assembly.

Artichoke is a characteristic crop of the Mediterranean area, recognized for its nutritional value and therapeutic properties due to the presence of bioactive components such as polyphenols, inulin, vitamins and minerals. Artichoke is mainly consumed after home and/or industrial processing, and the undersized heads, not suitable for the market, can be used for the recovery of bioactive compounds, such as polyphenols, for cosmetic applications. In this paper, the potential skin anti-age effect of a polyphenolic artichoke extract on endothelial cells was investigated. The methodology used was addressed to evaluate the antioxidant and anti-inflammatory activities and the improvement of gene expression of some youth markers. The results showed that the artichoke extract was constituted by 87% of chlorogenic, 3,5-O-dicaffeoylquinic, and 1,5-O-dicaffeoylquinic acids. The extract induced important molecular markers responsible for the microcirculation and vasodilatation of endothelial cells, acted as a potential anti-inflammatory agent, protected the lymphatic vessels from oxidative damage by ROS formation, and enhanced the cellular cohesion by reinforcing the tight junction complex. In addition, the artichoke extract, through the modulation of molecular pathways, improved the expression of genes involved in anti-ageing mechanisms. Finally, clinical testing on human subjects highlighted the enhancement by 19.74% of roughness and 11.45% of elasticity from using an artichoke extract cosmetic formulation compared to placebo cream.

Plants are sessile organisms and, in order to defend themselves against exogenous (a)biotic constraints, they synthesize an array of secondary metabolites which have important physiological and ecological effects. Plant secondary metabolites can be classified into four major classes: terpenoids, phenolic compounds, alkaloids and sulphur-containing compounds. These phytochemicals can be antimicrobial, act as attractants/repellents, or as deterrents against herbivores. The synthesis of such a rich variety of phytochemicals is also observed in undifferentiated plant cells under laboratory conditions and can be further induced with elicitors or by feeding precursors. In this review, we discuss the recent literature on the production of representatives of three plant secondary metabolite classes: artemisinin (a sesquiterpene), lignans (phenolic compounds) and caffeine (an alkaloid). Their respective production in well-known plants, i.e., Artemisia, Coffea arabica L., as well as neglected species, like the fibre-producing plant Urtica dioica L., will be surveyed. The production of artemisinin and caffeine in heterologous hosts will also be discussed. Additionally, metabolic engineering strategies to increase the bioactivity and stability of plant secondary metabolites will be surveyed, by focusing on glycosyltransferases (GTs). We end our review by proposing strategies to enhance the production of plant secondary metabolites in cell cultures by inducing cell wall modifications with chemicals/drugs, or with altered concentrations of the micronutrient boron and the quasi-essential element silicon.

Higher plants are the source of a wide array of bioactive compounds that support skin integrity and health. Hibiscus syriacus, family Malvaceae, is a plant of Chinese origin known for its antipyretic, anthelmintic, and antifungal properties. The aim of this study was to assess the healing and hydration properties of H. syriacus ethanolic extract (HSEE). We established a cell culture from Hibiscus syriacus leaves and obtained an ethanol soluble extract from cultured cells. The properties of the extract were tested by gene expression and functional analyses on human fibroblast, keratinocytes, and skin explants. HSEE treatment increased the healing potential of fibroblasts and keratinocytes. Specifically, HSEE significantly stimulated fibronectin and collagen synthesis by 16 and 60%, respectively, while fibroblasts contractility was enhanced by 30%. These results were confirmed on skin explants, where HSEE accelerated the wound healing activity in terms of epithelium formation and fibronectin production. Moreover, HSEE increased the expression of genes involved in skin hydration and homeostasis. Specifically, aquaporin 3 and filaggrin genes were enhanced by 20 and 58%, respectively. Our data show that HSEE contains compounds capable of stimulating expression of biomarkers relevant to skin regeneration and hydration thereby counteracting molecular pathways leading to skin damage and aging.

The term "oxidative stress" indicates a set of chemical reactions unleashed by a disparate number of events inducing DNA damage, lipid peroxidation, protein modification and other effects, which are responsible of altering the physiological status of cells or tissues. Excessive Reactive Oxygen Species (ROS) levels may accelerate ageing of tissues or induce damage of biomolecules thus promoting cell death or proliferation in dependence of cell status and of targeted molecules. In this context, new antioxidants preventing such effects may have a relevant role as modulators of cell homeostasis and as therapeutic agents. Following an approach of peptide libraries synthesis and screening by an ORACFL assay, we have isolated potent anti-oxidant compounds with well-defined structures. Most effective peptides are N-terminally trifluoroacetylated (CF3) and have the sequence tyr-tyr-his-pro or tyr-tyr-pro-his. Slight changes in the sequence or removal of the CF3 group strongly reduced antioxidant ability, suggesting an active role of both the fluorine atoms and of peptide structure. We have determined the NMR solution structures of the active peptides and found a common structural motif that could underpin the radical scavenging activity. The peptides protect keratinocytes from exogenous oxidation, thereby from potential external damaging cues, suggesting their use as skin ageing protectant and as cell surviving agents.

Skin ageing is a complex and progressive phenomenon, characterized by dramatic morphological changes of the cells, which results in wrinkle formation and loss of elasticity. Moreover, as skin ages, it looses the capacity to generate new young thrusting cells, since the stem cell reservoirs are not proliferating actively, contrarily to a young and healthy skin. Nevertheless cell ageing looks like an irreversible phenomenon, recently it was demonstrated that when aged cells are transplanted into a young tissue, they can “rejuvenate”, meaning that they recover their capacity to proliferate and fulfill their functional activities, suggesting that the ageing process mostly depends on the biological context where cells locate and on the number and types of chemical signals the cells perceive. On the basis of these studies, one important blood factor responsible of reversing several senescent phenotypes associated with aged cells was isolated and characterized, the Growth Differentiation Factor 11 (GDF11). In plants, somatic embryogenesis is the maximum expression of how cells have an unlimited capacity to repair themselves and generate young tissues, despite of their age. It is a natural phenomenon by which somatic cells turn back to a young phenotype and, by acquiring the ability to proliferate, develop into a zygotic-like structure, called somatic embryo, similar to the one contained in the seeds. On the basis of these studies, we investigated the role of GDF11 in skin cells and how its expression was affected by the treatment with an extract derived from the somatic embryos, developed from the plant species Lotus japonicus. We demonstrated that GDF11 expression and synthesis are reduced in aged fibroblasts and Lotus japonicus somatic embryo extract significantly increased the production of GDF11, with a consequent increase in the expression of other genes involved in cell longevity and rejuvenation.