The angico alcoholic extract (Anadenanthera colubrina var. cebil) induces the acceleration of wound healing in skin wounds of rats.
Skin wound healing is a complex process involving different events such as blood coagulation, inflammation, new blood vessels formation, and extracellular matrix deposition. These events can be observed by using histology techniques. However, the lack of the standardization of such parameters impacts on the reproducibility of results. Here, we describe a protocol to perform macroscopic and microscopic analyses of the events that occur during skin wound healing using the experimental model of excisional wounds in rats.
PURPOSE:To evaluate the effects of angico bark extract (Anadenanthera colubrina var. cebil) in the healing process of the skin of rats. METHODS:Twenty adult male rats were divided into four groups of five animals each, according to the respective postoperative days, as follow: G4, G7, G14 and G21. Each group received two incisions on skin and subcutaneous tissue in the right and left antimere of the thoracic region, separated by a distance of 2 cm. The right lesion was treated daily with saline and the left with the angico alcoholic extract (5%). At the end of each experimental period, the animals were euthanized and fragments of the wound area with the edges were removed, fixed in 10% formaldehyde solution and processed for paraffin embedding. Histological sections (5 µm of thickness) were stained with hematoxylin and eosin (HE), Gomori trichromic and picrosisirus red for morphological and morphometric analyses.Statistical analysis was done by ANOVA and Tukey-Kramer test (p<0.05). RESULTS:Morphological analysis showed larger fibroblasts and a higher concentration of collagen fibers in skyn wounds treated with the angico extract. Morphometric analysis demonstrated a significant increase in the number of fibroblasts at 7th and collagen in 7th and 14th days (p<0.01) in wounds treated with the angico extract. CONCLUSION:The angico alcoholic extract (Anadenanthera colubrina var. cebil) induces the acceleration of wound healing in skin wounds of rats.
Resumo -O objetivo deste trabalho foi avaliar morfometricamente a neoangiogênese de retalhos cutâneos subdérmicos em ratos tratados com óleo de copaíba (Copaifera langsdorffi i) em pomada a 10%. O delineamento experimental foi inteiramente casualizado, composto por três recursos farmacológicos. Foram utilizadas dez repetições para cada recurso, e cada animal foi considerado uma unidade experimental. Trinta ratos Wistar foram submetidos à elevação do retalho cutâneo dorsal subdérmico e distribuídos em três grupos: grupo controle absoluto, no qual os animais não receberam nenhum tratamento; grupo controle, no qual os animais receberam tratamento tópico diário com pomada com apenas veículo (glicerina e vaselina); e grupo tratado, no qual os animais foram tratados diariamente com óleo de copaíba em pomada a 10%. Os ratos foram tratados e observados por oito dias após o ato operatório. No oitavo dia de pós-operatório, realizou-se a análise macroscópica do retalho e foram coletados fragmentos das porções cranial, média e caudal do retalho cutâneo para análise histopatológica. A análise morfométrica mostrou diferença signifi cativa para o número de novos vasos sanguíneos nas partes média e caudal do retalho cutâneo no grupo tratado. O óleo de copaíba mostra-se efi ciente no aumento da neoangiogênese em retalhos cutâneos subdérmicos de ratos.Termos para indexação: Copaifera langsdorffi i, isquemia, neovascularização. Neoangiogenesis of skin fl aps in rats treated with copaiba oilAbstract -The aim of this work was to evaluate morphometric aspects of subdermic skin fl ap neoangiogenesis in rats treated with a 10% copaiba (Copaifera langsdorffi i) oil ointment. The experiment design was a completely randomized, with three pharmacological resources. Ten replicates were used for each resource, and each animal was considered one experimental unit. Thirty Wistar rats were submitted to subdermic dorsal skin fl ap elevation and distributed into three groups: absolute control group, in which the animals did not receive any treatment; control group, in which the animals received topical treatment only with vehicle (glycerin and petroleum jelly), and treated group, in which the animals were treated daily with a 10% copaiba oil ointment. The rats were treated and observed for eight days after surgery. On the eighth day, the skin fl ap underwent a macroscopic analysis and fragments were collected from its cranial, medial and caudal portions for histopathological analysis. The morphometric analysis showed signifi cant difference for proliferation of newly formed vessels in the middle and caudal portions of the skin fl ap in the treated group. Copaiba oil showed effi ciency in the increase of neoangiogenesis of subdermic skin fl aps in rats.
Purpose: To evaluate wound contraction and the concentration of mast cells in skin wounds treated with wild plum (Ximenia americana) essential oil-based ointment in rats. Methods: Sixty rats were submitted to two cutaneous wounds in the thoracic region, on the right and left antimeres. Thereon, they were divided into three groups: GX (wounds treated once a day with hydro alcoholic branch extract of Ximenia amerIcana), GP (wounds that received vehicle), and GC (wounds without product ap-plication). Wounds were measured immediately after the injury as well as 4, 7, 14 and 21 days post-topical application of the extract. At these days, five rats from each group were euthanatized. Thereafter, samples were fixed in 10% formalde-hyde and processed for paraffin embedding. Sections were stained with H.E, Masson's Trichrome and toluidine blue for morphological, morphometrical and histopathological analysis, under light microscopy. The degree of epithelial con-traction was measured and mast cell concentrations were also evaluated with an image analyzer (Image Pro-plus®software). Results: The extract treated group showed lower mast cell concentrations in the 4th day of lesion, as compared to GP (GX
This study aimed to perform phytochemical analysis and to test the antimicrobial activity of the crude hydroalcoholic extract obtained from the leaves of Sphagneticola trilobata. Classes of secondary metabolites present in the extract were identified through phytochemical screening using analytical thin-layer chromatography. Antimicrobial activity was evaluated by testing cultures of Staphylococcus aureus, S. epidermidis, Staphylococcus spp., Escherichia coli, Serratia marcescens, Enterococcus faecalis, Pseudomonas aeruginosa, Salmonella Typhimurium, and Klebsiella pneumoniae isolated from human skin and those of Staphylococcus spp. isolated from dog skin using the broth microdilution method. In the phytochemical screening, classes of anthracenic derivatives and mono-, sesqui-, and diterpenes were identified. Colorimetric analysis showed total phenol and total flavonoid contents of 21.7 ± 0.009 mg of gallic acid equivalents per gram of sample and 0.23 ± 0.005 mg of catechin equivalents per gram of sample, respectively. Microbiological analysis revealed that the hydroalcoholic extract of S. trilobata exhibited antimicrobial activity against cultures of Staphylococcus spp., E. coli, S. marcescens, and E. faecalis isolated from human skin and those of Staphylococcus spp. isolated from dog skin. Thus, crude hydroalcoholic extract of leaves of S. trilobata contained flavonoids and terpenoids as secondary metabolites, which contributed to its antimicrobial activity against skin bacteria isolated from different sources.
Purpose To evaluate the healing potential of the Ximenia americana hydroalcoholic extract in 10% cream in excisional wound models in rats. Methods Sixty male adults Wistar rats were submitted to skin and subcutaneous tissue surgery in the right and left thoracic regions, divided into three experimental groups: Standard submitted to treatment with only the base vehicle, Treated wounds treated with hydroalcoholic extract of X. americana applied on 10%, Lanette base and Control, untreated wounds. The treatment was performed daily and the wounds evaluated microscopically by the quantification of fibroblasts, collagen fibers and blood vessels. Results The histomorphometric analysis showed a significant increase in the number of fibroblasts, collagen fibers and blood vessels in the treated group. Conclusion The topical action of the cream based on Ximenia americana shows angiogenic effects and improves the replacement of collagen, suggesting its use for the development of herbal remedy in the treatment of cutaneous wound healing.
To evaluate the effects of rosemary leaf essential oil-based ointments on the healing of rat skin lesions. Methods: Sixty adult male rats, with dorsal excisional skin wounds made surgically under anesthesia, were divided into three groups (n = 20): Sham group (untreated wounds); control group (CG, wounds treated with vehicle); and essential oil (EO) treated group (wounds treated with essential oil-based ointments), administered topically once daily. Skin wounds were evaluated at 4, 7, 14, and 21 days after EO or vehicle treatments. Lesions were analyzed macroscopically for the contraction degree. Formalin-fixed paraffin-embedded sections of skin wounds were used for histopathological evaluation. Results: Macroscopic evaluation showed wounds edges with thin crust without firmness and yellowish color, along with an improvement in wound contraction in EO group when compared to the other groups. A reduced inflammatory reaction, along with newly formed small diameter capillaries and more organized and elongated collagen fibers, were more frequently observed in EO group than in the other groups. Moreover, blood vessel number and collagen fibers density were significantly higher in EO group. Conclusion: Skin lesion treatment with rosemary leaf essential oil-based ointments accelerates the initial stages of healing, reduces inflammation, and increases angiogenesis, collagen fibers density, and wound contraction in rats.
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