A novel engineered dermis for in vitro photodamage research.

The realization of biologically relevant human tissue equivalents as an in vitro model to investigate human diseases, as well as to test the efficacy or toxicity of novel compounds, is emerging as a new challenge in tissue engineering. Currently, the in vitro three-dimensional (3D) dermis model mainly involves the use of cells embedded in exogenous non-human matrices. However, such models feature biological and functional disparities with native dermis, therefore limiting their relevance to the in vivo situation. The purpose of this study was to provide a reliable endogenous human dermal equivalent (HDE) able to recapitulate the extracellular matrix (ECM) remodelling of the native dermis occurring after external damage. To this end, UVA irradiation was used to induce photodamage to both the HDE and to a fibroblast-populated collagen matrix. The photodamage was investigated at the cellular and ECM level and the results showed that, although a cellular response was detected in both systems, no ECM reorganization characteristic of the in vivo photo-aged dermis could be detected in the fibroblast-populated collagen matrix. In contrast in the HDE, the neosynthesized ECM recapitulated the characteristic ageing behaviour of the dermis found in vivo, in terms of collagen and hyaluronic acid synthesis as well as collagen organization remodelling. This study therefore demonstrates the role of the endogenous ECM in recapitulating in vitro the functionality of the human dermis and the proposed HDE as a novel tool for photoprotection trials. Copyright © 2016 John Wiley & Sons, Ltd.

Hibiscus syriacus Extract from an Established Cell Culture Stimulates Skin Wound Healing.

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.

An oil-soluble extract of Rubus idaeus cells enhances hydration and water homeostasis in skin cells.

OBJECTIVE: Raspberry plants, belonging to the species of Rubus idaeus, are known for their excellent therapeutic properties as they are particularly rich in compounds with strong antioxidant activity, which promote health and well-being of human cells. Besides their high content of phenolic compounds, Rubus plants are rich in oil-soluble compounds, which are also primary components of the hydrolipidic film barrier of the skin. As plant cell cultures represented a valuable system to produce interesting compounds and ingredients for cosmetic applications, we developed liquid suspension cultures from Rubus idaeus leaves and used them to obtain an active ingredient aimed at improving hydration and moisturization capacity in the skin. METHODS: Rubus idaeus cells, grown in the laboratory under sterile and controlled conditions as liquid suspension cultures, were processed to obtain an oil-soluble (liposoluble) extract, containing phenolic compounds and a wide range of fatty acids. The extract was tested on cultured keratinocytes and fibroblasts and then on the skin in vivo, to assess its cosmetic activities. RESULTS: When tested on skin cell cultures, the extract induced the genes responsible for skin hydration, such as aquaporin 3, filaggrin, involucrin and hyaluronic acid synthase, and stimulated the expression and the activity of the enzyme glucocerebrosidase, involved in ceramide production. Moreover, the liposoluble extract increased the synthesis of the extracellular matrix components in cultured fibroblasts and showed a remarkable skin-hydrating capacity when tested on human skin in vivo. CONCLUSIONS: Thanks to these activities, the Rubus idaeus liposoluble extract has several potential applications in skin care cosmetics: it can be used as hydrating and moisturizing ingredient in face and body lotions, and as anti-ageing product in face creams specifically designed to fight wrinkle formation.

Fish assemblage structure of the Ipanema River, a small lotic environment partially protected by a Conservation Unit in southeastern Brazil.

A study on the fish assemblage of the Ipanema River, a small affluent of the Tietê River basin in southeastern Brazil, was performed aiming to look for structural patterns of species diversity in small lowland lotic environments. Fish samplings were performed every two months from June 2003 to April 2004 at four sample sites located on the lower stretch of the river. Local assemblage showed to be species rich, with fifty-two species belonging to Characiformes (25 spp.), Siluriformes (19 spp.), Cyprinodontiformes (3 spp.), Gymnotiformes (2 spp.), Perciformes (2 spp.), and Synbranchiformes (1 sp.). Fish fauna was composed of small-sized species (<200 mm SL) and by individuals of medium (up to 400 mm SL) to large (more than 400 mm SL) sized species. The Ipanema River, such as other small lotic transitional environments in the upper Paraná River drainage, is considered important for conservation of fish fauna because they cover available habitats for persistent populations of small-sized species and for non-persistent individuals or shoals of medium and large-sized fish species, which occupy other habitats along their life-history (e. g. floodplains, oxbow lakes, main channel of great rivers). The importance of the Ipanema River basin for fish fauna conservation is also reinforced by the fact that it is located in a highly impacted region of southeastern Brazil, near the São Paulo metropolitan area.

Fish assemblage structure of the Ipanema River, a small lotic environment partially protected by a Conservation Unit in southeastern Brazil / Estrutura da assembleia de peixes do Rio Ipanema, um pequeno ambiente lotico parcialmente protegido por uma Unidade de Conservacao no Sudeste do Brasil

A study on the fish assemblage of the Ipanema River, a small affluent of the Tietê River basin in southeastern Brazil, was performed aiming to look for structural patterns of species diversity in small lowland lotic environments. Fish samplings were performed every two months from June 2003 to April 2004 at four sample sites located on the lower stretch of the river. Local assemblage showed to be species rich, with fifty-two species belonging to Characiformes (25 spp.), Siluriformes (19 spp.), Cyprinodontiformes (3 spp.), Gymnotiformes (2 spp.), Perciformes (2 spp.), and Synbranchiformes (1 sp.). Fish fauna was composed of small-sized species (<200 mm SL) and by individuals of medium (up to 400 mm SL) to large (more than 400 mm SL) sized species. The Ipanema River, such as other small lotic transitional environments in the upper Paraná River drainage, is considered important for conservation of fish fauna because they cover available habitats for persistent populations of small-sized species and for non-persistent individuals or shoals of medium and large-sized fish species, which occupy other habitats along their life-history (e. g. floodplains, oxbow lakes, main channel of great rivers). The importance of the Ipanema River basin for fish fauna conservation is also reinforced by the fact that it is located in a highly impacted region of southeastern Brazil, near the São Paulo metropolitan area.

Um estudo sobre a estrutura da assembleia de peixes do Rio Ipanema, um pequeno afluente da bacia do Rio Tietê, no Sudeste do Brasil, foi realizado com o objetivo de buscar padrões estruturais de diversidade de espécies em pequenos ambientes lóticos de planícies. Peixes foram coletados a cada dois meses, de junho de 2003 a abril de 2004, em quatro locais de amostragem situados no trecho inferior do rio. A assembleia de peixes se mostrou rica em espécies, com 52 espécies registradas, pertencentes a 40 gêneros, 19 famílias e às ordens Characiformes (25 spp.); Siluriformes (19 spp.); Cyprinodontiformes (3 spp.); Gymnotiformes (2 spp.); Perciformes (2 spp.), e Synbranchiformes (1 sp.). A fauna de peixes foi composta por espécies de pequeno porte (<200 mm CP) e por indivíduos de espécies que atingem porte médio (até 400 mm CP) e grande (mais de 400 mm CP). O Rio Ipanema, assim como outros pequenos ambientes lóticos de transição entre riachos e grandes rios na drenagem do Alto Rio Paraná, são considerados importantes para a conservação da ictiofauna porque representam habitats disponíveis para populações persistentes de espécies de pequeno porte e para indivíduos ou cardumes não persistentes de espécies de médio e grande porte, que podem ocupar outros habitats (por exemplo, as lagoas marginais, as planícies de inundação e as calhas principais de grandes rios), durante o transcorrer de sua vida. Esta importância para conservação também é ressaltada porque a Bacia do Rio Ipanema é localizada em uma região intensamente impactada do Sudeste do Brasil, próxima à área metropolitana de São Paulo.

In vivo and in vitro assessment of pathways involved in contrast media-induced renal cells apoptosis.

Contrast-induced nephropathy accounts for >10% of all causes of hospital-acquired renal failure, causes a prolonged in-hospital stay and represents a powerful predictor of poor early and late outcome. Mechanisms of contrast-induced nephropathy are not completely understood. In vitro data suggests that contrast media (CM) induces a direct toxic effect on renal tubular cells through the activation of the intrinsic apoptotic pathway. It is unclear whether this effect has a role in the clinical setting. In this work, we evaluated the effects of CM both in vivo and in vitro. By analyzing urine samples obtained from patients who experienced contrast-induced acute kidney injury (CI-AKI), we verified, by western blot and immunohistochemistry, that CM induces tubular renal cells apoptosis. Furthermore, in cultured cells, CM caused a dose-response increase in reactive oxygen species (ROS) production, which triggered Jun N-terminal kinases (JNK1/2) and p38 stress kinases marked activation and thus apoptosis. Inhibition of JNK1/2 and p38 by different approaches (i.e. pharmacological antagonists and transfection of kinase-death mutants of the upstream p38 and JNK kinases) prevented CM-induced apoptosis. Interestingly, N-acetylcysteine inhibited ROS production, and thus stress kinases and apoptosis activation. Therefore, we conclude that CM-induced tubular renal cells apoptosis represents a key mechanism of CI-AKI.