RESUMO
OBJECTIVE: Evaluation of the healing and toxicological effects of Copaifera duckei Dwyer oleoresin (CDO). MATERIALS AND METHODS: Rodents with skin lesions were divided into nine groups, including daily treatments with 1, 3 and 10% CDO, collagenase, antibiotic ointment and control groups, for 14 days. RESULTS: Treatment with 10% CDO reduced skin edema and hyperplasia, demonstrating anti-inflammatory effect of the oil. Reduction in the wound area was observed, indicating the healing effect of CDO. Histopathological analysis showed increases in angiogenesis and re-epithelialization in animals treated with the highest concentration. On the other hand, no alterations in ulcerations, inflammatory infiltrate, hemorrhage, congestion, degeneration, percentage of collagen fibers, number of cells stained with anti-macrophage migration inhibitory factor, or density of area stained with anti-collagen I and III were found. Toxicogenetic analysis revealed no differences in micronucleus frequencies or in the ratio of polychromatic erythrocytes to total erythrocytes between treated and negative control, demonstrating the absence of genotoxicity and cytotoxicity, respectively. There was no difference in levels of liver enzymes among groups, indicating the absence of hepatotoxicity. CONCLUSION: Formulations of CDO exerted beneficial effects on the stages of cutaneous wound healing and are promising options for the treatment of wounds.
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Skin wound healing is a highly complex event that involves different mediators at the cellular and molecular level. Lupeol has been reported to possess different biological activities, such as anti-inflammatory, antioxidant, antidiabetic, and in vitro wound healing properties, which motivated us to proceed with in vivo studies. We aimed to investigate the wound healing effect of lupeol-based cream for 3, 7, and 14 days. Wound excisions were induced on the thoraco-lumbar region of rats and topically treated immediately after injury induction. Macroscopic, histopathological, and immunohistochemical analyses were performed. Cytokine levels were measured by ELISA and gene expression was evaluated by real-time RT-qPCR. Our results showed a strong wound-healing effect of lupeol-based cream after 7 and 14 days. Lupeol treatment caused a reduction in proinflammatory cytokines (TNF-a, IL-1ß, and IL-6) and gene and protein NF-κB expression, and positively altered IL-10 levels, showing anti-inflammatory effects in the three treatment periods. Lupeol treatment showed involvement in the proliferative phase by stimulating the formation of new blood vessels, increasing the immunostaining of Ki-67 and gene expression, and immunolabeling of vascular endothelial growth factor (VEGF) and epidermal growth factor (EGF), and increasing gene expression of transforming growth factor beta-1 (TGF-ß1) after seven days of treatment. Lupeol was also involved in the tissue regeneration phase by increasing the synthesis of collagen fibers noted in the three treatment periods analyzed. Our findings suggest that lupeol may serve as a novel therapeutic option to treat cutaneous wounds by regulating mechanisms involved in the inflammatory, proliferative, and tissue-remodeling phases.
Assuntos
Anti-Inflamatórios/uso terapêutico , Colágeno/metabolismo , Citocinas/metabolismo , Fármacos Dermatológicos/uso terapêutico , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Antígeno Ki-67/metabolismo , NF-kappa B/metabolismo , Triterpenos Pentacíclicos/uso terapêutico , Fitoterapia , Cicatrização/efeitos dos fármacos , Administração Cutânea , Animais , Anti-Inflamatórios/administração & dosagem , Anti-Inflamatórios/farmacologia , Fármacos Dermatológicos/administração & dosagem , Fármacos Dermatológicos/farmacologia , Regulação da Expressão Gênica/efeitos dos fármacos , Inflamação , Peptídeos e Proteínas de Sinalização Intercelular/genética , Antígeno Ki-67/genética , Masculino , NF-kappa B/genética , Neovascularização Fisiológica/efeitos dos fármacos , Triterpenos Pentacíclicos/administração & dosagem , Triterpenos Pentacíclicos/farmacologia , RNA Mensageiro/biossíntese , RNA Mensageiro/genética , Ratos , Ratos Wistar , Regeneração/efeitos dos fármacos , Fenômenos Fisiológicos da Pele/efeitos dos fármacosRESUMO
INTRODUÇÃO: O cuidado do paciente com feridas é um constante desafio na prática clínica do enfermeiro. As coberturas oclusivas e não oclusivas são utilizadas no tratamento de feridas e existe uma lacuna de conhecimento sobre a influência dessas indicações no processo de cicatrização. OBJETIVO: O objetivo do presente estudo foi avaliar a influência de coberturas oclusivas sobre a cicatrização de feridas cutâneas excisionais em camundongos C57BL/6. MATERIAIS E MÉTODOS: Trata-se de um estudo translacional pré-clínico, comparativo e controlado. Os camundongos foram divididos em três grupos experimentais e o tratamento das feridas excisionais deles foram classificados como: animais que receberam oclusão com hidrocolóide (HD), animais que receberam oclusão com filme transparente de poliuretano (FT) e animais sem oclusão (SO). Os grupos foram avaliados quanto: a taxa de fechamento das feridas, o infiltrado de neutrófilos e macrófagos, dosagem da citocina TNF-α, o fator de crescimento angiogênico VEGF, a expressão protéica de HIF-1α e, histologicamente, o desenvolvimento de novos vasos sanguíneos. As cicatrizes das feridas foram analisadas por juízes independentes quanto à apresentação estética no décimo quarto dia de segmento. RESULTADO: No terceiro dia do processo de cicatrização, foi possível observar, macroscopicamente, que o fechamento das feridas cutâneas excisionais foi acelerado nos animais cuja ferida foi ocluída, utilizando-se tanto filme transparente quanto o hidrocolóide. Ainda no período de três dias, observou-se diminuição nos níveis da citocina pró-inflamatória TNF-α nas feridas ocluídas com hidrocolóide quando comparada com os grupos SO e FT. O acúmulo de neutrófilos também diminuiu, significativamente, ao terceiro dia de cicatrização no grupo tratado com hidrocolóide, quando comparado aos grupos SO e FT, respectivamente. O conteúdo dos macrófagos apresentou níveis significativamente mais altos ao terceiro dia no grupo FT quando comparado a SO. Ao sexto dia de estudo, evidenciaram-se dosagens significativamente aumentadas de macrófagos no grupo HD, contrastando com os grupos SO e FT. Os níveis de VEGF apresentaramse expressivamente aumentados ao sexto dia de cicatrização, nos animais cujas feridas estavam ocluídas com filme transparente de poliuretano e hidrocolóide. Houve aumento dos níveis do fator indutor de hipóxia 1-alfa nos animais submetidos a oclusão quando comparados aos sem oclusão. A análise qualitativa do reparo em cicatriz das feridas permitiu afirmar que houve concordância moderada e significativa entre os juízes no grupo HD e concordância substancial e significativa nas cicatrizes dos animais SO. CONSIDERAÇÕES FINAIS: Considerando-se que a resposta inflamatória é um processo imunológico fundamental para o processo de reparo de feridas, quando ocorre de forma controlada, sugere-se que a oclusão modula a resposta inflamatória. Tal achado é evidenciado por meio da inibição parcial da citocina pró-inflamatória TNF- α, da redução de acúmulo de neutrófilos e dos níveis mais altos de macrófagos. A oclusão das feridas não evidenciou aumento quantitativo de capilares em relação ao grupo sem oclusão.(AU)
INTRODUCTION: The care of the wounded patient is a constant challenge in the nurse's clinical practice. Occlusive and non-occlusive coverages are used to treat wounds and there is a lack of knowledge about the impact of these indications on the healing process. OBJECTIVE: The objective of the present study was to evaluate the influence of occlusive coatings on the cicatrization of cutaneous excisional wounds in C57BL / 6 mice. MATERIALS AND METHODS: It is a pre-clinical, comparative and controlled translational study. The mice were divided into three experimental groups and the treatment of excisional wounds of these were classified as: animals that received occlusion with hydrocolloid (HD), animals that received occlusion with transparent polyurethane film (FT) and animals without occlusion (SO). The groups were evaluated for: wound closure rate, neutrophil and macrophage infiltration, TNF-α cytokine dosing, VEGF angiogenic growth factor, HIF-1α protein expression and, histologically, the development of new blood vessels. Wound scars were analyzed by independent judges regarding the aesthetic presentation on the fourteenth day of the segment. RESULTS: On the third day of the healing process it was possible to observe, macroscopically, that the closure of excisional skin wounds was accelerated in animals whose wound was occluded using both transparent and hydrocolloid films. Also, in the three-day period, there was a decrease in TNF-α proinflammatory cytokine levels in the hydrocolloid occluded wounds when compared to the SO and FT groups. Neutrophil accumulation also significantly decreased on the third day of healing in the group treated with hydrocolloid occlusion when compared to the SO and FT groups, respectively. Macrophage content had significantly higher levels, on day 3, in the FT group, when compared to OS. On the sixth day of the study, significantly increased dosages of macrophages were detected in the HD group, contrasting with the SO and FT groups. VEGF levels were expressively increased on the sixth day of healing in animals whose wound was occluded with transparent polyurethane film and hydrocolloid film. There was an increase in levels of the factor inducing 1-alpha hypoxia in the animals submitted to occlusion when compared to those without occlusion. The qualitative analysis of the wound healing repair allowed to affirm that there was moderate and significant agreement, between the judges, in the HD group and substantial and significant concordance in the scars of the SO animals. FINAL CONSIDERATIONS: Considering that the inflammatory response is a fundamental immunological process for the wound repair process, when it occurs in a controlled way, it is suggested that the occlusion modulates the inflammatory response. Such a finding is evidenced by partial inhibition of the proinflammatory cytokine TNF-α, reduction of neutrophil accumulation, and higher levels of macrophages. The wounds occlusion did not show a quantitative increase of capillary vessels in relation to the group without occlusion.(AU)
Assuntos
Animais , Camundongos , Cicatrização , Curativos Hidrocoloides , Curativos Oclusivos , Dissertação Acadêmica , Inflamação/terapiaRESUMO
A cicatrização de feridas é um processo que requer a interação de várias células da derme e epiderme. O objetivo deste trabalho foi avaliar qual o momento da aplicação das células das ADSCs em feridas cutâneas agudas que faria diferença na cicatrização nos primeiros sete dias da lesão. As células-tronco foram isoladas do tecido adiposo de camundongos C57Bl/6 GFP+. Para tanto, foram utilizados 49 camundongos C57Bl/6, divididos em quatro grupos: grupo I (GI/controle; n=14); grupo II (GII; n=14): ADSCs injetadas no d0; grupo III (GIII; n=14): ADSCs injetadas no terceiro dia; e Grupo IV (GIV; n=7): ADSCs injetadas no quinto dia. As avaliações clínicas ocorreram nos dias zero, três, cinco e sete, e as histopatológicas nos dias cinco e sete. Na metodologia proposta, foi observado que o uso de ADSCs aumenta a vascularização, a formação de tecido de granulação, a colagenização e incrementa o número de folículos pilosos em apenas sete dias de avaliação. Além disso, o momento da aplicação das células não repercutiu diferenças significativas nas fases inflamatória e proliferativa do processo de cicatrização das feridas cutâneas.(AU)
Wound healing is a process that requires the interaction of various cells in the dermis and epidermis. The aim of this study was to evaluate the action of ADSCs in the treatment of acute wounds in order to understand if application time of the cells results in a difference in healing the first seven days of injury. The stem cells were isolated from adipose tissue of C57BL / 6 mice GFP +. Thus, we used 49 mice C57BL / 6 divided into four groups: Group I (GI / control, n=14); Group II (GII; n=14): ADSCs injected to the d0; Group III (GIII; n=14): ADSCs injected on the 3rd day, and Group IV (GIV; n=7): ADSCs injected day 5(d5). Clinical evaluations were performed on days 0, 3, 5 and 7 and the histopathology on days 5 and 7. In the proposed methodology, the use of ADSCs increased vascularization, formation of granulation tissue, collagen deposition and increases the number of hair follicles in just seven days of evaluation. In addition, the time of application of the cells did not affect significant differences in the inflammatory and the proliferative phase of wound healing skin.(AU)
Assuntos
Animais , Camundongos , Células-Tronco , Cicatrização/fisiologia , Tecido Adiposo , Inflamação/veterináriaRESUMO
A cicatrização de feridas é um processo que requer a interação de várias células da derme e epiderme. O objetivo deste trabalho foi avaliar qual o momento da aplicação das células das ADSCs em feridas cutâneas agudas que faria diferença na cicatrização nos primeiros sete dias da lesão. As células-tronco foram isoladas do tecido adiposo de camundongos C57Bl/6 GFP+. Para tanto, foram utilizados 49 camundongos C57Bl/6, divididos em quatro grupos: grupo I (GI/controle; n=14); grupo II (GII; n=14): ADSCs injetadas no d0; grupo III (GIII; n=14): ADSCs injetadas no terceiro dia; e Grupo IV (GIV; n=7): ADSCs injetadas no quinto dia. As avaliações clínicas ocorreram nos dias zero, três, cinco e sete, e as histopatológicas nos dias cinco e sete. Na metodologia proposta, foi observado que o uso de ADSCs aumenta a vascularização, a formação de tecido de granulação, a colagenização e incrementa o número de folículos pilosos em apenas sete dias de avaliação. Além disso, o momento da aplicação das células não repercutiu diferenças significativas nas fases inflamatória e proliferativa do processo de cicatrização das feridas cutâneas.(AU)
Wound healing is a process that requires the interaction of various cells in the dermis and epidermis. The aim of this study was to evaluate the action of ADSCs in the treatment of acute wounds in order to understand if application time of the cells results in a difference in healing the first seven days of injury. The stem cells were isolated from adipose tissue of C57BL / 6 mice GFP +. Thus, we used 49 mice C57BL / 6 divided into four groups: Group I (GI / control, n=14); Group II (GII; n=14): ADSCs injected to the d0; Group III (GIII; n=14): ADSCs injected on the 3rd day, and Group IV (GIV; n=7): ADSCs injected day 5(d5). Clinical evaluations were performed on days 0, 3, 5 and 7 and the histopathology on days 5 and 7. In the proposed methodology, the use of ADSCs increased vascularization, formation of granulation tissue, collagen deposition and increases the number of hair follicles in just seven days of evaluation. In addition, the time of application of the cells did not affect significant differences in the inflammatory and the proliferative phase of wound healing skin.(AU)
Assuntos
Animais , Camundongos , Células-Tronco , Cicatrização/fisiologia , Tecido Adiposo , Inflamação/veterináriaRESUMO
ABSTRACT Wound healing is a process that requires the interaction of various cells in the dermis and epidermis. The aim of this study was to evaluate the action of ADSCs in the treatment of acute wounds in order to understand if application time of the cells results in a difference in healing the first seven days of injury. The stem cells were isolated from adipose tissue of C57BL / 6 mice GFP +. Thus, we used 49 mice C57BL / 6 divided into four groups: Group I (GI / control, n=14); Group II (GII; n=14): ADSCs injected to the d0; Group III (GIII; n=14): ADSCs injected on the 3rd day, and Group IV (GIV; n=7): ADSCs injected day 5(d5). Clinical evaluations were performed on days 0, 3, 5 and 7 and the histopathology on days 5 and 7. In the proposed methodology, the use of ADSCs increased vascularization, formation of granulation tissue, collagen deposition and increases the number of hair follicles in just seven days of evaluation. In addition, the time of application of the cells did not affect significant differences in the inflammatory and the proliferative phase of wound healing skin.
RESUMO A cicatrização de feridas é um processo que requer a interação de várias células da derme e epiderme. O objetivo deste trabalho foi avaliar qual o momento da aplicação das células das ADSCs em feridas cutâneas agudas que faria diferença na cicatrização nos primeiros sete dias da lesão. As células-tronco foram isoladas do tecido adiposo de camundongos C57Bl/6 GFP+. Para tanto, foram utilizados 49 camundongos C57Bl/6, divididos em quatro grupos: grupo I (GI/controle; n=14); grupo II (GII; n=14): ADSCs injetadas no d0; grupo III (GIII; n=14): ADSCs injetadas no terceiro dia; e Grupo IV (GIV; n=7): ADSCs injetadas no quinto dia. As avaliações clínicas ocorreram nos dias zero, três, cinco e sete, e as histopatológicas nos dias cinco e sete. Na metodologia proposta, foi observado que o uso de ADSCs aumenta a vascularização, a formação de tecido de granulação, a colagenização e incrementa o número de folículos pilosos em apenas sete dias de avaliação. Além disso, o momento da aplicação das células não repercutiu diferenças significativas nas fases inflamatória e proliferativa do processo de cicatrização das feridas cutâneas.
RESUMO
CONTEXT: The quantification of total collagen is of major importance in a wide range of research areas, including the study of cutaneous wound healing and new drugs trials. OBJECTIVE: The total collagen content in skin biopsies was compared by biochemical hydroxyproline assay and by two computer-aided histomorphometric analyses of histological sections. MATERIALS AND METHODS: Two methods were used to evaluate collagen formation: the hydroxyproline assay, as the gold standard and histomorphometric image analysis of the filled areas by corresponding stained collagen fibres, using picrosirius and Gomori's trichrome staining. The image analyses were determined by digital densitometry recognition using computer-aided ImageJ software. One-way ANOVA, simple linear regression and ANCOVA were applied for the statistical analysis and correlation. RESULTS: In a simple linear regression analysis carried out on the 14th day period after the induction of skin injury, three techniques, picrosirius red (F = 33.57, p = 0.00), Gomori's trichrome (F = 81.61, p = 0.00) and hydroxyproline content (F = 16.85, p = 0.00) were able to detect collagen production. After scale adjustment, there were no significant differences among either the slopes (F = 1.17, p = 0.32) or the intercepts (F = 0.69, p = 0.51) of the estimated regression lines. It seems that a highly significant correlation exists between the histomorphometrical analysis and hydroxyproline assay. DISCUSSION AND CONCLUSION: The morphometric analysis proved to be adequate and can be used as a simple, rapid, low-cost technology for evaluating total collagen in cutaneous wound specimens, compared with the gold standard hydroxyproline assay.
Assuntos
Colágeno/análise , Pele/lesões , Cicatrização/fisiologia , Animais , Hidroxiprolina/análise , Processamento de Imagem Assistida por Computador , Masculino , Ratos , Ratos Wistar , Pele/químicaRESUMO
BACKGROUND: The overproduction of reactive oxygen species (ROS) and exacerbated inflammatory response are the main events that impair healing of pressure ulcers. Therefore, olive oil may be a good alternative to improve the healing of these chronic lesions due to its anti-inflammatory and antioxidant properties. OBJECTIVE: This study investigated the effect of olive oil administration on wound healing of pressure ulcers in mice. METHODS: Male Swiss mice were daily treated with olive oil or water until euthanasia. One day after the beginning of treatment, two cycles of ischemia-reperfusion by external application of two magnetic plates were performed in skin to induced pressure ulcer formation. RESULTS: The olive oil administration accelerated ROS and nitric oxide (NO) synthesis and reduced oxidative damage in proteins and lipids when compared to water group. The inflammatory cell infiltration, gene tumor necrosis factor-α (TNF-α) expression and protein neutrophil elastase expression were reduced by olive oil administration when compared to water group. The re-epithelialization and blood vessel number were higher in the olive oil group than in the water group. The olive oil administration accelerated protein expression of TNF-α, active transforming growth factor-ß1 and vascular endothelial growth factor-A when compared to water group. The collagen deposition, myofibroblastic differentiation and wound contraction were accelerated by olive oil administration when compared to water group. CONCLUSION: Olive oil administration improves cutaneous wound healing of pressure ulcers in mice through the acceleration of the ROS and NO synthesis, which reduces oxidative damage and inflammation and promotes dermal reconstruction and wound closure.
Assuntos
Inflamação/tratamento farmacológico , Óxido Nítrico/metabolismo , Azeite de Oliva/uso terapêutico , Úlcera por Pressão/tratamento farmacológico , Espécies Reativas de Oxigênio/metabolismo , Pele/metabolismo , Cicatrização/efeitos dos fármacos , Administração Cutânea , Animais , Colágeno/metabolismo , Modelos Animais de Doenças , Humanos , Masculino , Camundongos , Azeite de Oliva/administração & dosagem , Reepitelização/efeitos dos fármacos , Pele/irrigação sanguínea , Pele/efeitos dos fármacos , Fator de Crescimento Transformador beta1/metabolismo , Fator de Necrose Tumoral alfa/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismoRESUMO
Two oil blends (sunflower/canola oils 85/15 (BL1) and canola/linseed oils 70/30 (BL2)), were prepared and enzymatically interesterified to be applied to surgically-induced wounds in rats. Following surgery, the animals were submitted to the Treatment with Physiological Saline (TPS) (control group), Blends (TBL), and Structured Lipids (TSL). The control group (TPS) received physiological saline solution for 15 days. In TBL, BL1 was administered during the inflammation phase (days 0-3) and BL2 in the tissue formation and remodeling phase (days 4-15). In TSL, Structured Lipid 1 (SL1) and Structured Lipid 2 (SL2) were used instead of BL1 and BL2, respectively. The aim of this study was to compare wound closure evolution among rats treated with the blends or structured lipids versus control rats treated with physiological saline. The wound healing process was evaluated by measuring the wound areas along the treatments and the concentrations of cytokines. An increase in the areas of wounds treated with the blends and structured lipids in the inflammatory phase was observed, followed by a steeper closure curve compared to wounds treated with physiological saline. The changes observed during the inflammatory phase suggest a potential therapeutic application in cutaneous wound healing which should be further investigated...
Duas misturas de óleos vegetais (girassol/canola 85/15 (BL1) e canola/linhaça, 70/30 (BL2) foram preparadas e interesterificadas por via enzimática para serem aplicadas em feridas induzidas cirurgicamente em ratos. Após a cirurgia, os animais foram submetidos ao tratamento com soro fisiológico (TPS) (grupo controle), tratamento com as misturas (TBL) e tratamento com os lipídios estruturados (TSL). O grupo controle (TPS) recebeu soro fisiológico por 15 dias. Em TBL, BL1 foi administrada durante a fase de inflamação (dias 0-3) e BL2 na fase de formação de tecido e remodelação (dias 4-15). Em TSL, os lipídios estruturados SL1 e SL2 foram usados em vez de BL1 e BL2, respectivamente. O objetivo deste estudo foi avaliar a evolução do fechamento das feridas dos grupos de ratos tratados com as misturas ou lipídios estruturados em comparação com os ratos do grupo controle, tratados com soro fisiológico. O processo de cicatrização das feridas foi avaliado através da medição das áreas das feridas ao longo dos tratamentos e pela determinação das concentrações de citocinas. Observou-se aumento das áreas das feridas tratadas com as misturas e os lipídios estruturados na fase inflamatória, seguida por um fechamento acentuado de feridas comparado com o tratamento com solução salina. As mudanças observadas durante a fase inflamatória sugerem uma potencial aplicação terapêutica na cicatrização de feridas cutâneas, fazendo-se necessárias investigações posteriores...
Assuntos
Masculino , Ratos , Infecção dos Ferimentos/tratamento farmacológico , Óleos de Plantas/efeitos adversos , Óleos de Plantas/farmacologia , Análise de Variância , Cicatrização , Citocinas/análiseRESUMO
The α2-adrenoceptors regulate the sympathetic nervous system, controlling presynaptic catecholamine release. However, the role of the α2-adrenoceptors in cutaneous wound healing is poorly understood. Mice lacking both the α2A/α2C-adrenoceptors were used to evaluate the participation of the α2-adrenoceptor during cutaneous wound healing. A full-thickness excisional lesion was performed on the dorsal skin of the α2A/α2C-adrenoceptor knockout and wild-type mice. Seven or fourteen days later, the animals were euthanized and the lesions were formalin-fixed and paraffin-embedded or frozen. Murine skin fibroblasts were also isolated from α2A/α2C-adrenoceptor knockout and wild-type mice, and fibroblast activity was evaluated. The in vivo study demonstrated that α2A/α2C-adrenoceptor depletion accelerated wound contraction and re-epithelialization. A reduction in the number of neutrophils and macrophages was observed in the α2A/α2C-adrenoceptor knockout mice compared with wild-type mice. In addition, α2A/α2C-adrenoceptor depletion enhanced the levels of nitrite and hydroxyproline, and the protein expression of transforming growth factor-ß and vascular endothelial growth factor. Furthermore, α2A/α2C-adrenoceptor depletion accelerated blood vessel formation and myofibroblast differentiation. The in vitro study demonstrated that skin fibroblasts isolated from α2A/α2C-adrenoceptor knockout mice exhibited enhanced cell migration, α-smooth muscle actin _protein expression and collagen deposition compared with wild-type skin fibroblasts. In conclusion, α2A/α2C-adrenoceptor deletion accelerates cutaneous wound healing in mice.