In Vitro and In Vivo Evaluation of Chitosan/HPMC/Insulin Hydrogel for Wound Healing Applications.

Treatment of chronic wounds is challenging, and the development of different formulations based on insulin has shown efficacy due to their ability to regulate oxidative stress and inflammatory reactions. The formulation of insulin with polysaccharides in biohybrid hydrogel systems has the advantage of synergistically combining the bioactivity of the protein with the biocompatibility and hydrogel properties of polysaccharides. In this study, a hydrogel formulation containing insulin, chitosan, and hydroxypropyl methyl cellulose (Chi/HPMC/Ins) was prepared and characterized by FTIR, thermogravimetric, and gel point analyses. The in vitro cell viability and cell migration potential of the Chi/HPMC/Ins hydrogel were evaluated in human keratinocyte cells (HaCat) by MTT and wound scratch assay. The hydrogel was applied to excisional full-thickness wounds in diabetic mice for twenty days for in vivo studies. Cell viability studies indicated no cytotoxicity of the Chi/HPMC/Ins hydrogel. Moreover, the Chi/HPMC/Ins hydrogel promoted faster gap closure in the scratch assay. In vivo, the wounds treated with the Chi/HPMC/Ins hydrogel resulted in faster wound closure, formation of a more organized granulation tissue, and hair follicle regeneration. These results suggest that Chi/HPMC/Ins hydrogels might promote wound healing in vitro and in vivo and could be a new potential dressing for wound healing.

An older diabetes-induced mice model for studying skin wound healing.

Advances in wound treatment depend on the availability of animal models that reflect key aspects of human wound healing physiology. To this date, the accepted mouse models do not reflect defects in the healing process for chronic wounds that are associated with type two diabetic skin ulcers. The long term, systemic physiologic stress that occurs in middle aged or older Type 2 diabetes patients is difficult to simulate in preclinical animal model. We have strived to incorporate the essential elements of this stress in a manageable mouse model: long term metabolic stress from obesity to include the effects of middle age and thereafter onset of diabetes. At six-weeks age, male C57BL/6 mice were separated into groups fed a chow and High-Fat Diet for 0.5, 3, and 6 months. Treatment groups included long term, obesity stressed mice with induction of diabetes by streptozotocin at 5 months, and further physiologic evaluation at 8 months old. We show that this model results in a severe metabolic phenotype with insulin resistance and glucose intolerance associated with obesity and, more importantly, skin changes. The phenotype of this older age mouse model included a transcriptional signature of gene expression in skin that overlapped that observed with elderly patients who develop diabetic foot ulcers. We believe this unique old age phenotype contrasts with current mice models with induced diabetes.

Accelerative action of topical piperonylic acid on mice full thickness wound by modulating inflammation and collagen deposition.

Epidermal growth factor (EGF) promotes cell growth, proliferation, and survival in numerous tissues. Piperonylic acid, a metabolite present in peppers (Piper nigrum L. and Piper longum L.), can bind to the epidermal growth factor receptor (EGFR) and induce an intracellular signaling cascade leading to the transcription of genes responsible for these actions, especially in keratinocytes. These cells are fundamental in maintaining cutaneous homeostasis and are the first to be damaged in the case of a wound. Thus, we hypothesized that piperonylic acid improves wound healing. C57BL6/J male mice were submitted to dorsal skin wounds caused by a 6 mm punch and treated topically with piperonylic acid or vehicle. The wounds were evaluated macro- and microscopically, and tissue samples were collected for immunofluorescence and real-time PCR analyses on days 6, 9 and 19 post-injury. Topical piperonylic acid improved wound healing from day 6 post-injury until closure. This phenomenon apparently occurred through EGFR activation. In addition, piperonylic acid modulated the gene expression of interleukin (Il)-6, il-1ß, tumor necrosis factor (Tnf)-α, il-10, monocyte chemoattractant protein (Mcp)-1 and insulin-like growth factor (Igf)-1, which are important for the healing process. By day 19 post-injury, the new tissue showed greater deposition of type I collagen and a morphology closer to intact skin, with more dermal papillae and hair follicles. We conclude that piperonylic acid may be a viable option for the treatment of skin wounds.

Glutamic acid promotes hair growth in mice.

Glutamic acid is the main excitatory neurotransmitter acting both in the brain and in peripheral tissues. Abnormal distribution of glutamic acid receptors occurs in skin hyperproliferative conditions such as psoriasis and skin regeneration; however, the biological function of glutamic acid in the skin remains unclear. Using ex vivo, in vivo and in silico approaches, we showed that exogenous glutamic acid promotes hair growth and keratinocyte proliferation. Topical application of glutamic acid decreased the expression of genes related to apoptosis in the skin, whereas glutamic acid increased cell viability and proliferation in human keratinocyte cultures. In addition, we identified the keratinocyte glutamic acid excitotoxic concentration, providing evidence for the existence of a novel skin signalling pathway mediated by a neurotransmitter that controls keratinocyte and hair follicle proliferation. Thus, glutamic acid emerges as a component of the peripheral nervous system that acts to control cell growth in the skin. These results raise the perspective of the pharmacological and nutritional use of glutamic acid to treat skin diseases.

Hypothalamic Microglial Heterogeneity and Signature under High Fat Diet-Induced Inflammation.

Under high-fat feeding, the hypothalamus atypically undergoes pro-inflammatory signaling activation. Recent data from transcriptomic analysis of microglia from rodents and humans has allowed the identification of several microglial subpopulations throughout the brain. Numerous studies have clarified the roles of these cells in hypothalamic inflammation, but how each microglial subset plays its functions upon inflammatory stimuli remains unexplored. Fortunately, these data unveiling microglial heterogeneity have triggered the development of novel experimental models for studying the roles and characteristics of each microglial subtype. In this review, we explore microglial heterogeneity in the hypothalamus and their crosstalk with astrocytes under high fat diet-induced inflammation. We present novel currently available ex vivo and in vivo experimental models that can be useful when designing a new research project in this field of study. Last, we examine the transcriptomic data already published to identify how the hypothalamic microglial signature changes upon short-term and prolonged high-fat feeding.

Bioactive Fatty Acids in the Resolution of Chronic Inflammation in Skin Wounds.

Significance: Optimal skin wound healing is crucial for maintaining tissue homeostasis, particularly in response to an injury. The skin immune system is under regulation of mediators such as bioactive lipids and cytokines that can initiate an immune response with controlled inflammation, followed by efficient resolution. However, nutritional deficiency impacts wound healing by hindering fibroblast proliferation, collagen synthesis, and epithelialization, among other crucial functions. In this way, the correct nutritional support of bioactive lipids and of other essential nutrients plays an important role in the outcome of the wound healing process. Recent Advances and Critical Issues: Several studies have revealed the potential role of lipids as a treatment for the healing of skin wounds. Unsaturated fatty acids such as linoleic acid, α-linolenic acid, oleic acid, and most of their bioactive products have shown an effective role as a topical treatment of chronic skin wounds. Their effect, when the treatment starts at day 0, has been observed mainly in the inflammatory phase of the wound healing process. Moreover, some of them were associated with different dressings and were tested for clinical purposes, including pluronic gel, nanocapsules, collagen films and matrices, and polymeric bandages. Therefore, future research is still needed to evaluate these dressing technologies in association with different bioactive fatty acids in a wound healing context. Future Directions: This review summarizes the main results of the available clinical trials and basic research studies and provides evidence-based conclusions. Together, current data encourage the use of bioactive fatty acids for an optimal wound healing resolution.

Topical Insulin Modulates Inflammatory and Proliferative Phases of Burn-Wound Healing in Diabetes-Induced Rats.

The healing time of burn wounds depends on surface area and depth of the burn and associated comorbidities. Diabetes mellitus (DM) causes delays in the healing process by extending the inflammatory phase. Treatment with topical insulin can improve the inflammatory phase, restore metabolic dysregulation, and modulate impaired cellular signaling in burn wounds. The objective of this study was to evaluate markers of the inflammatory and proliferative phases of second-degree burns after topical insulin treatment in diabetic rats. Type I DM was induced with streptozotocin in male Wistar rats. The animals' backs were shaved and subjected to thermal burning. Rats were randomized into two groups: control diabetic (DC) and insulin diabetic (DI). At Days 7 and 14 postburn, rats were euthanized, and wound-tissue sections were evaluated by hematoxylin and eosin, Weigert, and Verhöeff staining, immunohistochemistry-paraffin, and enzyme-linked immunosorbent assay. A significant increase in reepithelialization was seen on Days 7 and 14 in DI versus DC rats. On Day 7, interleukin (IL)-1ß, IL-6, monocyte chemotactic protein (MCP)-1, and F4/80 expression were increased in DI versus DC rats. On Day 14, MCP-1 expression was decreased and F4/80 increased in DI versus DC rats. On Days 7 and 14, Ki-67, transforming growth factor-ß1, vascular endothelial growth factor expression, and formation of elastic fibers were increased in DI versus DC rats. Topical insulin modulates burn-wound healing in diabetic animals by balancing inflammation and promoting angiogenesis and formation of elastic fibers.

Psychometric Performance of the Brazilian Version of the Diabetes Distress Scale in Patients With Diabetes Mellitus Type 2.

BACKGROUND AND PURPOSE: Assessing the stress of patient with diabetes requires reliable and valid instruments. This study evaluated the measurement properties of the Brazilian version of the Diabetes Distress Scale (B-DDS). METHODS: This cross-sectional study enrolled 139 patients with Type 2 diabetes mellitus (DM2) receiving outpatient treatment. Reliability and construct validity were estimated through convergent validity and confirmatory factor analysis. RESULTS: Evidence of reliability was obtained-Cronbach alpha = .87 and intraclass correlation coefficient (ICC) = .93. Significant positive correlations of moderate-to-strong magnitudes were observed between the dimensions of the DDS and the total score of the PAID; the confirmatory factor analysis supported the four dimensions of the original instrument. CONCLUSIONS: The B-DDS is reliable and valid for evaluation of the stress related to diabetes mellitus.

Effect of atorvastatin on wound healing in rats.

Skin-wound healing is a complex and dynamic biological process involving inflammation, proliferation, and remodeling. Recent studies have shown that statins are new therapeutical options because of their actions, such as anti-inflammatory and antioxidant activity, on vasodilation, endothelial dysfunction and neoangiogenesis, which are independent of their lipid-lowering action. Our aim was to investigate the effect of atorvastatin on tissue repair after acute injury in healthy animals. Rats were divided into four groups: placebo-treated (P), topical atorvastatin-treated (AT), oral atorvastatin-treated (AO), topical and oral atorvastatin-treated (ATO). Under anesthesia, rats were wounded with an 8-mm punch in the dorsal region. Lesions were photographed on Days 0, 1, 3, 7, 10, 12, and 14 post-injury and samples taken on Days 1, 3, 7, and 14 for protein-expression analysis of insulin receptor substrate (IRS)-1, phosphatidylinositol 3-kinase (PI3K), protein kinase B (Akt), glycogen synthase kinase (GSK)-3, endothelial nitric oxide synthase (eNOS), vascular endothelial growth factor (VEGF), extracellular signal-regulated kinase (ERK), interleukin (IL)-10, IL-1ß, IL-6, and tumor necrosis factor (TNF)-α. Upon macroscopic examination, we observed significant reductions of lesion areas in groups AT, AO, and ATO compared to the P group. Additionally, AT and AO groups showed increased expression of IRS-1, PI3K, Akt, GSK-3, and IL-10 on Days 1 and 3 when compared with the P group. All atorvastatin-treated groups showed higher expression of IRS-1, PI3K, Akt, GSK-3, IL-10, eNOS, VEGF, and ERK on Day 7. On Days 1, 3, and 7, all atorvastatin-treated groups showed lower expression of IL-6 and TNF-α when compared with the P group. We conclude that atorvastatin accelerated tissue repair of acute lesions in rats and modulated expressions of proteins and cytokines associated with cell-growth pathways.

Papaína-ureia como agente desbridante: revisão de literatura / Papína-urea como agente del desbridamiento: revisión de la literatura / Papain-urea a debriding agent: literature review

A remoção do tecido inviável da ferida é um passo importante para o processo de cicatrização. O desbridamento enzimático é um dos métodos utilizados pela equipe de enfermagem para tal fim, consistindo na aplicação de enzimas exógenas no leito da ferida, a fim de obter a degradação do tecido necrosado, sem prejuízo do viável. Dentre os produtos disponíveis para tal desbridamento, a associação papaína-ureia apresenta resultados satisfatórios. Entretanto, ainda é pouco utilizada no Brasil. Assim, o presente estudo é uma revisão de literatura, cujo objetivo foi identificar artigos científicos sobre o uso da papaína-ureia como agente desbridante em feridas. A busca se deu nas seguintes bases de dados: PubMed, Scopus, Web of Science, Cochrane, LILACS e SciELO. O período considerado foi de 2000 a 2012, utilizando os descritores: desbridamento, papaína-ureia, papaína, ureia e suas respectivas versões em inglês. De acordo com a literatura, o uso de papaína-ureia promove dissolução do tecido não viável e melhora a aparência da lesão: ocorre redução de edema, eritema, odor e exsudato. Favorece, assim, o processo de granulação e epitelização. Entretanto, as evidências científicas ainda são poucas e mais estudos precisam ser realizados.

The removal of infeasible tissue from a wound is an important step to the healing process. Enzymatic debridement is one of the methods used by nursing staff for this, what consisits in the application of exogenous enzymes on wound in order to obtain necrotic tissue degradation without prejudice to feasible tissues. Among the available products for such debridement, the association of papain and urea produces satisfactory results. However, it is not widely used in Brazil. Therefore, this literature review aimed to identify papers on using papain-urea as debriding agent for wounds. The search took place in the following databases: PubMed, Scopus, Web of Science, Cochrane, LILACS and SciELO. The considered period was from 2000 to 2012, using the following keywords in Portuguese and English: debridement, papain-urea, papain, and urea. According to literature, the use of papain-urea causes dissolution of non-viable tissue and improves the appearance of lesions: it decreases edema, erythema, odor, and exudate. Thus, it favors granulation and tissue epithelialization. However, there are few scientific evidences regarding this issue and more studies must be conducted.Keywords: Debridement. Papain. Urea. Wound healing. Nursing care..

La eliminación de tejido no viable de la herida es un paso importante en el proceso de cicatrización. Desbridamiento enzimático es uno de los métodos utilizados por el personal de enfermería, que consiste en la aplicación de las enzimas exógenas en la herida con el fin de obtener la degradación de tejido necrótico, sin daño para viables. Entre los productos disponibles para tal desbridamiento, la asociación de la papaína y urea produce resultados satisfactorios. Sin embargo, no se utiliza ampliamente en Brasil. Por lo tanto, este estudio es una revisión de la literatura que tuvo como objetivo identificar artículos sobre el uso de papaína-urea como agente de desbridamiento para las heridas. La búsqueda se realizó en las siguientes bases de datos: PubMed, Scopus, Web of Science, Cochrane, LILACS y SciELO. El período considerado fue 2000-2012, utilizando las palabras clave en Portugués y Inglés: desbridamiento, papaína-urea, papaína y urea. Según la literatura, aplicación de la papaína-urea promueve disolución del tejido no viable, así como mejora la aparencia de la lesión: produce reducción de edema, eritema, olor y exudado. Por lo tanto favorece el proceso de granulación y epitelización. Pero, las evidencias científicas son aún pocas y más estudios necesitan ser realizados.

Nursing diagnoses of diabetic patient medical charts: a descriptive study

Aim: identifying taxonomy II nursing diagnoses from the North American Nursing Diagnoses Association International using nursing records associated with outpatient diabetic treatment. Method: a descriptive and retrospective study. Data obtained from 35 patients' medical charts, using a tool devised by the authors and analyzed using relative and absolute frequencies. Result: from eight diagnoses, three of the following showed up in more than 50% of the samples: Ineffective health maintenance; Imbalanced nutrition - more than body requirements; Sedentary lifestyle. Discussion: The findings show that these three diagnoses relate to a very important and unique issue in treating diabetic patients: the difficulty on the part of patients to adhere to treatment and self-care. Conclusion: The findings show that nursing diagnoses are useful with regard to identifying phenomena that require attention in specific treatment contexts, as well as acting as a guide in terms of the assistance offered.

Diabetes mellitus e o processo de cicatrização cutânea / Diabetes mellitus and the process of cutaneous healing / Diabetes mellitus y el proceso de cicatrización cutánea

O objetivo desta atualização foi abordar o mecanismo molecular e celular do processo de cicatrização de feridas cutâneas em indivíduos com diabetes mellitus. O processo de reparo tecidual em indivíduos com diabetes mellitus é lentificado, a produção excessiva de Espécies Reativas de Oxigênio, diminuição do Óxido Nítrico, diminuição da reposta aos Fatores de Crescimento e das proteínas da via de sinalização da insulina estão envolvidos neste processo. O sucesso no tratamento de feridas de difícil cicatrização depende do conhecimento dos fatores que interferem neste processo, que aliado à prática clínica trará ao enfermeiro subsídios para prevenção, intervenções e escolha de coberturas adequadas para o cuidado e tratamento de feridas em pacientes com diabetes mellitus.

The objective of this updating study was to address the molecular and cellular process of the healing of cutaneous wounds in individuals with diabetes mellitus. The process of tissue repair in individuals with diabetes mellitus is slowed down; the excessive production of Reactive oxygen species, the reduction of Nitric Oxide, the reduction in the response to Growth Factors and the reduction in the proteins of the insulin signalling route are all involved in this process. Success in treating wounds which are having difficulty in healing depends on knowledge of the factors which interfere in this process, which, allied with clinical practice, provides the nurse with support for prevention, interventions and choice of appropriate dressings for the care and treatment of wounds in patients with diabetes mellitus.

La finalidad de esta actualización fue plantear el mecanismo molecular y celular del proceso de cicatrización de heridas cutáneas en individuos con diabetes mellitus. El proceso de restauración tecidual en individuos con diabetes mellitus es lentificado, la producción excesiva de Especies Reactivas de Oxigenio, reducción del Óxido Nítrico, reducción de la respuesta a los Factores de Crecimiento y de las proteínas de la vía de señalización de la insulina hacen parte de este proceso. El éxito en el tratamiento de heridas de dificil cicatrización depende del conocimiento de los factores que interfieren en este proceso, lo cual junto a la práctica clínica traerá al enfermero subsidios para prevención, intervenciones y elección de medidas adecuadas para el cuidado y tratamiento de heridas en pacientes con diabetes mellitus.