Metformin-induced alterations in gills of the freshwater fish Astyanax lacustris (Lütken, 1875) detected by histological and scanning electron microscopy.

The antidiabetic drug metformin is widely prescribed and found in different concentrations in the environment around the world, raising concern about potential impacts on aquatic life. Analyses of the effects of exposure of biological models to aquatic contaminants are important for assessing pollution effects on fish health. The gills of fishes represent primary targets of disturbance by pollutants, mainly because of the large surface of the respiratory epithelium and the high perfusion rate, which both help the entry of pollutants into this tissue. In this context, the aim of this work was to use gill histological analyses biomarkers to evaluate the toxicity of metformin on aquatic environmental systems, by means of chronic exposure for 90 days of Astyanax lacustris (lambari), an ecologically important neotropical species that can be used as an environmental bioindicator. Histopathological analyses were performed using Light and Scanning Electron Microscopy. The main changes were lamellar fusion, telangiectasia hyperplasia and disappearance of microridges. The morphological changes observed possibly interfere with the gill physiology, indicating an unfavorable situation to the presence of metformin in the water, pointing to a concern that metformin may pose a risk to Astyanax lacustris and likely to other fish species, compromising the dynamics of the aquatic ecosystem as a whole. Graphical abstract.

Evaluation of a multiple microemulsion from Trichilia catigua extract and the percutaneous penetration through skin by Phase-Resolved photoacoustic spectroscopy.

Emulsion systems have been a breakthrough in cosmetic products, providing performance and effectiveness of products that use this technological strategy for drug delivery systems. In this sense, the microemulsion of the multiple emulsion W/O/W type containing a standardized extract of Trichilia catigua with high levels of polyphenols and antioxidants has great potential for cosmetic use. The aim of this study was to evaluate the formulations safety through the analysis of toxicity, comedogenicity, and histopathology in rabbits and apply the Phase-Resolved Photoacoustic Spectroscopy method to determine the formulation percutaneous penetration through the skin. The ex vivo experiments were performed in the ears of albino New Zealand rabbits treated twice a day for 14 days. The results of histological, hematological, and blood chemistry showed that the formulations are safe. Histopathological analysis showed no tissue reaction in any of the analyzed organs (liver and kidneys), confirming the absence of toxicity. Histological analysis showed that the formulations with extract of T. catigua demonstrated mild-moderately comedogenic and acanthosis compared to the control group. Inflammatory reactions, erythema, and desquamation were not observed in treated and controls animals. The phase-resolved photoacoustic spectroscopy method showed the penetration of the developed formulations throughout the rabbit's skin, identifying their absorption bands at the dermal side of the skin. In conclusion, the results of this study provide a step towards the application of the developed natural antioxidant encapsulated in a multiple microemulsion for skincare, concerned with the physical, chemical, and biological properties of the formulation.

Hydrogel containing (1 → 6)-ß-D-glucan (lasiodiplodan) effectively promotes dermal wound healing.

A hydrogel containing exocellular (1 → 6)-ß-D-glucan (lasiodiplodan, LAS) was developed and its wound healing potential was evaluated. ß-Glucans have attracted much interest by the cosmetic industry sector because of their bioactive and functional properties and in promoting skin health. In the present work an ß-glucan was studied as a healing biomaterial that has not hitherto been reported in the scientific literature. LAS produced by the ascomycete Lasiodiplodia theobromae MMPI was used in the formulation of a healing hydrogel. Physicochemical and microbiological quality parameters, antioxidant potential and stability of the formulation was evaluated. FTIR, thermal analysis and SEM techniques were also employed in the characterization. Wistar rats were used as a biological model to investigate the wound healing potential. Histological analyses of cutaneous tissue from the dorsal region were conducted after 4, 7, 10 and 14 days of treatment, and evaluated re-epithelialization, cell proliferation and collagen production. Physicochemical stability, microbiological quality and antioxidant potential, especially in relation to its ability to scavenge hydroxyl radicals were found. The hydrogel stimulated cell re-epithelialization and proliferation during all days of the treatment, and stimulated an increase of collagen fibers. Lasiodiplodan showed immunomodulatory activity in wound healing and this biomacromolecule could be an alternative compound in wound care.

Stem bark extract of Poincianella pluviosa incorporated in polymer film: Evaluation of wound healing and anti-staphylococcal activities.

Poincianella pluviosa has already been described as capable of healing skin wounds. In an attempt to prolong contact of the drug with the wound, it was proposed in this study to evaluate wound healing using a crude extract (CE) of P. pluviosa incorporated in carboxymethylcellulose polymer films. The chromatographic profile of the semipurified fraction of P. pluviosa was evaluated by ultra-high performance liquid chromatography (UHPLC), confirming the compounds gallic acid, geraniin, and ellagic acid. The films were evaluated for their physical and mechanical properties, water vapor permeability, moisture absorption capacity, and FTIR spectroscopy. For in vivo experiments, wounds were made on the back of rats and treated daily for 4, 7, 10, or 14 days with film containing CE or control film. At the end of each period, skin permeation analysis and histological analysis were made using re-epithelialisation, cell proliferation, and collagen formation. Statistical significance was determined by GraphPad Prism using t test and Mann-Whitney test. Anti-staphylococcal activity was evaluated with standard strains of Staphylococcus aureus, methicillin-resistant, and coagulase negative. It was demonstrated that the presence of CE in the films increased the capacity to absorb water and decreased resistance and permeability. The CE of the film permeated the skin, reaching the dermis and was able to influence re-epithelisation, cell proliferation, and collagen formation. Satisfactory results were observed against S. aureus strains, particularly coagulase negative. Films with CE of P. pluviosa can be an alternative in the wound healing, protecting against opportunistic infections and giving comfort to the patient.

Ação cicatrizante de plantas medicinais: um estudo de revisão / Healing action of medicinal plants: a review

Plantas Medicinais são utilizadas desde a antiguidade para tratar uma série de doenças desempenhando um papel fundamental na saúde mundial. O uso da fitoterapia é muitas vezes o único recurso terapêutico de algumas comunidades. A Organização Mundial da Saúde reconhece a fitoterapia como uma alternativa de tratamento viável com baixo custo e recomenda o levantamento, identificação, estímulo e orientação do uso das plantas medicinais que possuem eficácia e segurança terapêutica comprovada. Nos últimos anos, os fitoterápicos ganharam espaço importante na reparação de tecidos, obtendo a partir deles formulações que agem no tratamento das feridas. O presente estudo teve por objetivo analisar pesquisas envolvendo plantas medicinais e seu poder cicatrizante. Foram utilizados dados de bases online, dos últimos dez anos, utilizando como descritores: "feridas, cicatrização, plantas medicinais". Foram analisados os nomes científico e popular das plantas, a parte da planta que foi utilizada para os experimentos, tipo de extração e utilização, classes das substâncias e efeito na cicatrização. As plantas pesquisadas foram: Aloe vera (L.) Burm. f. (Babosa), Coronopu didymus (Mastruz), Tabebuia avellanedae (Ipê roxo), Arnica Montana L. (Arnica), Orbignya phalerata (Babaçu), Stryphnodendron adstringens Martius (Barbatimão), Caesalpinia ferrea Martius (Jucá), Chenopodium ambrosioides L. (Erva de Santa Maria), Triticum vulgare (Trigo), Tabernaemontana catharinensis (Jasmim), Calendula officinalis (Calêndula) e Helianthus annus (Girassol), Catharanthus roseus L. (Vinca Rosea) e Schinus terebinthifolius Raddi (Aroeira). Assim, pode-se observar que todas as plantas estudadas no artigo favoreceram o processo de cicatrização por meio de diferentes atividades, e que essas atividades estão relacionadas com as diversas classes de metabólitos

Medicinal plants have been used to treat a number of diseases since ancient times, and play a key role in human health. In recent decades, modern medicine has made great advances, but plants still contribute to the development of medicines. The use of herbal medicine is often the only therapeutic resource available in some communities. The World Health Organization recognizes herbal therapy as a viable treatment alternative at a low cost and recommends the collection, identification, stimulation and guidance of the use of medicinal plants that have presented efficacy and safety therapeutics. In recent years, herb therapy has gained important space in the repair of tissues through formulations that act in the treatment of wounds. The aim of this study was to analyze studies involving medicinal plants and their healing potential. Data from online databases from the past ten years were analyzed, using the descriptors: "wounds, healing, medicinal plants". The scientific and popular names of plants, the part of the plant used, type of extraction, therapeutic use, classes of substances and effect on wound healing were analyzed. The plants studied were: Aloe vera (L.) Burm. f. (Babosa), Coronopu didymus (lesser swine cress), Tabebuia avellanedae (pink ipe), Arnica Montana L. (wolf's bane), Orbignya phalerata (Babassu), Stryphnodendron adstringens Martius (Barbatimão), Caesalpinia ferrea Martius (Brazilian ironwood), Chenopodium ambrosioides L. (wormseed), Triticum vulgare (wheat), Tabernaemontana catharinensis (Jasmin), Calendula officinalis (Calendula) and Helianthus annus (sunflower), Catharanthus roseus L. (Madagascar periwinkle) and, Schinus terebinthifolius Raddi (Brazilian peppertree). Thus, the results showed that all the studied plants favored the wound healing process through different mechanism. These activities may be related to the different classes of substances

Pharmacognostic evaluation, and development and validation of a HPLC-DAD technique for gallocatechin and epigallocatechin in rhizomes from Limonium brasiliense

ABSTRACT Limonium brasiliense (Boiss.) Kuntze, Plumbaginaceae, is a plant from the southern coast of Brazilian that is used for the treatment of premenstrual syndrome, menstrual disorders and genito-urinary infections. The aim of the present study was to determine the quality control parameters for rhizomes collected during different periods by pharmacopoeial and non-pharmacopoeial methods, and to develop and validate a HPLC-DAD method for quantitative control of marker substances. The measured parameters were: granulometric analysis (d50 = 0.21–0.48 mm), loss on drying (11.1–12.4%), total ash (4.9–5.7%), dry residue by extraction with acetone:water (7:3, v/v) (30.6–39.5%), total polyphenol content (8.5–15.8%), and chromatographic fingerprint by HPLC and TLC. Besides, the acetone:water (7:3, v/v) extraction solvent in combination with a turbo-extractor, yielded the crude extract with a significant increase in tannins (F4,20 = 37.0, p < 0.001). The antioxidant potential of the crude acetone:water (7:3, v/v) extract, as well as the ethyl acetate and water fractions obtained after the partition process was evaluated by DPPH and the results were, respectively: IC50 6.87, 5.91, and 6.92 µg/ml. The validation parameters for the HPLC-DAD method showed adequate specificity, precision and accuracy. The gallo- and epigallocatechin contents were, respectively, 0.8–2.7% and 1.2–2.2%. These data contribute to analysis of the pharmacognostic quality control of the commonly used part from this species.

Larvicidal Activity against Aedes aegypti and Chemical Characterization of the Inflorescences of Tagetes patula.

The crude acetone extract (CAE) of defatted inflorescences of Tagetes patula was partitioned into five semipurified fractions: n-hexane (HF), dichloromethane (DF), ethyl acetate (EAF), n-butanol (BF), and aqueous (AQF). BF was fractionated by reversed-phase polyamide column chromatography, obtaining 34 subfractions, which were subjected to HSCCC, where patuletin and patulitrin were isolated. CAE and the fractions BF, EAF, DF, and AQF were analyzed by LC-DAD-MS, and patuletin and patulitrin were determined as the major substances in EAF and BF, respectively. BF was also analyzed by HPLC and capillary electrophoresis (CE), and patulitrin was again determined to be the main substance in this fraction. CAE and the semipurified fractions (750, 500, 300, 100, and 50 mg/L) were assayed for larvicidal activity against Aedes aegypti, with mortality rate expressed as percentage. All fractions except AQF showed insecticidal activity after 24 h exposure of larvae to the highest concentration. However, EAF showed the highest activity with more than 50% reduction in larval population at 50 mg/L. The insecticidal activity observed with EAF might have been due to the higher concentration of patuletin present in this fraction.

Enhanced Cutaneous Wound Healing In Vivo by Standardized Crude Extract of Poincianella pluviosa.

Wound healing is a complex process that involves several biological events, and a delay in this process may cause economic and social problems for the patient. The search continues for new alternative treatments to aid healing, including the use of herbal medicines. Members of the genus Caesalpinia are used in traditional medicine to treat wounds. The related species Poincianella pluviosa (DC.) L.P. Queiroz increases the cell viability of keratinocytes and fibroblasts and stimulates the proliferation of keratinocytes in vitro. The crude extract (CE) from bark of P. pluviosa was evaluated in the wound-healing process in vivo, to validate the traditional use and the in vitro activity. Standardized CE was incorporated into a gel and applied on cutaneous wounds (TCEG) and compared with the formulation without CE (Control) for 4, 7, 10, or 14 days of treatment. The effects of the CE on wound re-epithelialization; cell proliferation; permeation, using photoacoustic spectroscopy (PAS); and proteins, including vascular endothelial growth factor (VEGF), superoxide dismutase 2 (SOD-2) and cyclooxygenase 2 (COX-2) were evaluated. The TCEG stimulated the migration of keratinocytes at day 4 and proliferation on the following days, with a high concentration of cells in metaphase at 7 days. Type I collagen formed more rapidly in the TCEG. PAS showed that the CE had permeated through the skin. TCEG stimulated VEGF at day 4 and SOD-2 and COX-2 at day 7. The results suggest that the CE promoted the regulation of proteins and helped to accelerate the processes involved in healing, promoting early angiogenesis. This led to an increase in the re-epithelialized surface, with significant mitotic activity. Maturation of collagen fibers was also enhanced, which may affect the resistance of the extracellular matrix. PAS indicated a correlation between the rate of diffusion and biological events during the healing process. The CE from P. pluviosa appears promising as an aid in healing.

Development and characterization of multiple emulsions for controlled release of Trichilia catigua (Catuaba) extract.

Considering the antioxidant activity of the Trichilia catigua extract (TCE), the aim of the current study was to develop and characterize W/O/W multiple emulsions containing different vegetable oils as a platform to deliver a TCE. The extract displayed antioxidant activity (IC50) of 4.59 µg/mL and total phenol content (TPC) of 50.84%. Formulations were prepared by the phase-inversion emulsification method and analyzed for morphological appearance, pH, conductivity, droplet size and distribution, content of active, rheological properties, in vitro release, skin permeation, and stability. Formulations prepared with canola oil were selected and displayed regular morphology, mean diameter 2.77 µm (without TCE), 3.07 µm with 0.5% and 3.23 µm with 1.0% TCE. Rheometry (flow) showed pseudoplastic behavior with minimal thixotropy for both systems. TCE could be released from emulsions containing 1.0% and 0.5% TCE in a controlled manner for 16 and 23 h, respectively. The emulsions allowed good retention of TCE in the skin (stratum corneum, epidermis, and dermis). In a 180-d assessment of accelerated chemical stability, TPC was more reduced for the emulsions at 40 °C; other parameters remained stable. Multiple emulsions containing TCE were developed, exhibited good characteristics, and may be considered for future investigations as anti-aging formulations for the skin.

Acute toxicity and long-term safety evaluation of the crude extract from rhizomes of Limonium brasiliense in mice and rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Limonium brasiliense (Boiss.) Kuntze, Plumbaginaceae, popularly known as baicuru, has been used in folk medicine to treat menstrual cramps and to regulate menstrual periods. However, little is known about its safety. This study evaluated the safety through in vivo tests of the acute, long-term, and liver toxicity, and the mutagenicity of the crude extract (CE) from rhizomes of L. brasiliense. MATERIALS AND METHODS: The acute toxicity was assessed in Swiss mice, and the chronic toxicity in Wistar rats. Male and female mice received the CE orally in single doses of 1.0, 2.0, 3.0, 4.0, or 5.0 g/kg. Clinical changes and mortality rate were used as parameters to assess the acute toxicity. In the long-term evaluation, male and female Wistar rats were treated orally with daily doses of the CE (50, 100, or 200 mg/kg) for 90 days. Assessments of weight, behavior and food intake, urinalysis, biochemical and hematological analyses, as well as macro- and microscopic observations of several organs were performed. The redox state of the liver was evaluated as a means of investigating the liver toxicity, and the micronucleus test to assess mutagenicity was also performed. RESULTS: Evaluation of acute toxicity indicated no apparent clinical change in the animals; the LD50 was 4.8 g/kg. Evaluation after 90 days administration showed that the CE, even in higher doses than are considered therapeutic, appeared to be safe. The micronucleus test demonstrated a low mutagenic potential for the CE. CONCLUSION: Our results showed that treatment with the CE from L. brasiliense caused low or no toxicity, as assessed using these doses and evaluation methods.

Morpho-anatomical study of rhizome of Limonium brasiliense

AbstractLimonium brasiliense (Boiss.) Kuntze, Plumbaginaceae, is an herb popularly known as guaicuru, guaicurá or baicuru. The species inhabits salt marshes from the coastal region of southern Brazil, including Rio de Janeiro, to Uruguay and Argentina. Although widely used in folk medicine in the state of Rio Grande do Sul to treat genitourinary infections and to regulate menstrual periods, L. brasiliense has been little studied. The present morpho-anatomical study was undertaken to resolve some doubts in the literature as to the nature of the part of the plant that is used for medicinal purposes, a true rhizome or a root. The morpho-anatomical characteristics were analyzed with the aid of light and scanning electron microscopy. The botanical material was characterized as a rhizome with internodes that are evident in the younger but not the older portions. Microscopic analysis revealed the presence of a multilayered periderm with a cortex, ray parenchyma, and pith, formed by collenchyma tissue with abundant intercellular spaces in the outer portions of the cortex, responsible for the rigidity of the body, and with walls impregnated with phenolic compounds. The vascular bundles are collateral with elliptical to elongated cells, and with few conducting and sclerenchymal elements. Groups of sclereids are dispersed through the cortex and pith. These morpho-anatomical characteristics define the structure as a rhizome.

Stryphnodendron adstringens: Clarifying Wound Healing in Streptozotocin-Induced Diabetic Rats.

Diabetes mellitus is a serious public health problem in which a major complication is impaired wound healing. Among the strategies developed to foster tissue repair is the use of medicinal plants. The bark of Stryphnodendron adstringens, which is popularly used as an aid in wound healing, has a documented effect on wound repair in normal rats. This study evaluated the healing action of the crude extract of S. adstringens in diabetic rats, and its chemical content. Compounds present in the crude extract were characterized by mass spectrometry. In diabetic rats (streptozotocin 35 mg/kg), two wounds made in the skin were treated daily for 4, 7, 10, and 14 days with gel containing 1 % crude extract or with base gel. Histological analyses involved the measurement of the length and thickness of the re-epithelialized surface, quantification of the number of cells in mitosis, and types I and III collagen fibers. Also, cutaneous permeation by photoacoustic spectroscopy, and the expression of cyclooxygenase-2 and vascular endothelial growth factor by Western blot were assessed. The crude extract fingerprint showed masses indicating proanthocyanidins. The crude extract mainly stimulated cell migration and proliferation of keratinocytes at the beginning of the treatment in addition to stimulating the replacement of type III collagen fibers by type I collagen fibers at 10 and 14 days. The photoacoustic spectroscopy technique showed that the gel containing 1 % of crude extract permeated through the skin to the dermis, where the crude extract was found. Vascular endothelial growth factor was stimulated after 7 days of treatment with the crude extract and cyclooxygenase-2 at 4, 7, and 10 days. The crude extract of S. adstringens acted in tissue repair in wounds in diabetic rats by stimulating the production of collagen fibers at the wound site. The crude extract favored the formation of a more organized extracellular matrix and filled the entire extent of the wound, and also fostered the upregulation of cyclooxygenase-2 and vascular endothelial growth factor, which are essential to this process. These crude extract actions in diabetic wounds are probably due to the presence of proanthocyanidins.