Estudo de três modelos de predição de gravidade em uma unidade de terapia intensiva especializada no atendimento de pacientes com extensas queimaduras / Study of three severity prediction models in an intensive care unit specialized in the care of patients with extensive burns

OBJETIVO: Não existe na literatura consenso indicando o melhor índice mortalidade a ser aplicado para pacientes vítimas de grandes queimaduras. Contudo, os índices de gravidade podem determinar o quadro apresentado pelo paciente, além de avaliar a probabilidade de mortalidade e morbidade a partir do quadro patológico, possibilitando uma melhor compreensão sobre a gravidade da doença e efetividade do tratamento. O objetivo é avaliar se o índice de gravidade clínica fornecido por três escalas de prognóstico (LODS, APACHE II e Charlson) reflete a situação clínica dos pacientes nas primeiras 24 horas posteriores a sua admissão na UTI de queimados. MÉTODO: Trata-se de um estudo documental com análise retrospectiva utilizando dados de 225 pacientes internados em UTI especializada em tratamentos de grandes queimados. Os valores registrados no prontuário nas primeiras 24 horas de internação foram aplicados nos escores LODS, APACHE II e índice de comorbidade de Charlson. Os pacientes foram divididos em grupo 1 (alta, 229 pacientes) e grupo 2 (óbito, 26 pacientes). RESULTADOS: Como resultado, foi obtida diferença estatística entre os dois grupos no escore de LODS e APACHE II, havendo uma média de pontuação superior no grupo 2. Também houve diferença estatística significante em relação aos parâmetros renais e pulmonares no escore de LODS, predição da mortalidade do escore APACHE II e superfície corporal queimada. CONCLUSÕES: Este estudo indica que a porcentagem de superfície corporal queimada, a função pulmonar e renal são preditores significantes de morte para pacientes queimados. Os índices calculados pelo APACHE II e o LODS se mostraram úteis para uso junto a esta população.

OBJECTIVE: There is no consensus in the literature indicating the best mortality rate to be applied to patients suffering from major burns. However, the severity indexes can determine the condition presented by the patient, in addition to evaluating the probability of mortality and morbidity from the pathological condition, allowing a better understanding of the severity of the disease and the effectiveness of the treatment. The aim is to assess whether the clinical severity index provided by three prognostic scales (LODS, APACHE II and Charlson) reflects the clinical situation of patients in the first 24 hours after their admission to the burn ICU. METHODS: This is a study with a retrospective analysis using data from 225 patients admitted to an ICU specializing in the treatment of major burns. The values recorded in the medical records in the first 24 hours of hospitalization were applied to the LODS, APACHE II and Charlson comorbidity index scores. Patients were divided into group 1 (discharge, 229 patients) and group 2 (death, 26 patients). RESULTS: As a result, a statistical difference was obtained between the two groups in the LODS and APACHE II scores, with an average of higher scores in group 2. There was also a statistically significant difference in relation to renal and pulmonary parameters in the LODS score, predicting the mortality of the APACHE II score and burned body surface. CONCLUSIONS: This study indicates that the percentage of body surface burned, lung and kidney function are significant predictors of death for burn patients. The indices calculated by APACHE II and the LODS proved to be useful for use with this population.

COVID-19: como proceder na prática da cirurgia plástica no Brasil. O que sabemos até agora? / COVID-19: how to proceed with the practice of plastic surgery in Brazil. What do we know right now?

COVID-19 (coronavirus disease, descrito em 2019) é uma doença infecciosa causada pelo coronavírus da síndrome respiratória aguda grave 2 (SARS-CoV-2). A maioria dos casos confirmados é leve ou assintomático, mas os casos mais graves podem evoluir para pneumonia grave com insuficiência respiratória e morte. Atualmente ocorre, no Brasil, um cenário de aumento exponencial de casos, dificultando a identificação da fonte de contágio. Ainda não podemos precisar quando ocorrerá o pico do surto de COVID-19 em nosso país ou quando os números de novos contaminados e óbitos começarão a diminuir. Nesse momento, o mais importante é a proteção contra um vírus do qual não se conhece todos os detalhes sobre contágio, transmissão e tratamento. A pandemia impactou e modificou a assistência médica, principalmente das especialidades cirúrgicas, onde o atendimento presencial é fundamental e não pode ser substituído integralmente pela telemedicina. Assim, o objetivo desta revisão foi compilar aspectos teóricos e práticos referentes à pandemia COVID-19 e seu impacto na rotina da atividade da cirurgia plástica. São propostos protocolos para retomada de nossas rotinas, analisando a experiência de países em fase mais adiantada da pandemia.

COVID-19 (coronavirus disease, described in 2019) is an infectious disease caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Most confirmed cases are mild or asymptomatic, but the most severe cases can progress to severe pneumonia with respiratory failure and death. In Brazil, there is a scenario of an exponential increase in cases, making it challenging to identify the source of contagion. We cannot yet specify when the peak of the COVID-19 outbreak will occur in our country or when the numbers of new contaminants and deaths will begin to decrease. So, the most important thing is protection against a virus for which all the details about contagion, transmission, and treatment are not known. The pandemic impacted and modified medical care, especially for surgical specialties, where face-to-face care is essential and cannot be replaced entirely by telemedicine. Thus, this review aimed to compile theoretical and practical aspects regarding the pandemic COVID-19 and its impact on plastic surgery activity routine. Protocols are proposed for resuming our routines, analyzing countries' experience at an advanced stage of the pandemic.

Effects of Hydrogel With Enriched Sodium Alginate in Wounds of Diabetic Patients.

Objective of this study was to evaluate the efficacy of the autolytic debridement promoted by hydrogel with sodium alginate enriched with fatty acids and vitamins A and E in the healing of foot wounds in diabetic patients. A clinical study was conducted at an outpatient clinic of medical specialties. The sample comprised 8 patients supervised for a 3-month period, from April to July 2017, by means of a clinical history, photographic record, planimetry, and classification of the wound severity by the Pressure Ulcer Scale for Healing (PUSH) system. Of the 8 patients supervised, 1 dropped out and 7 were followed up for 12 weeks. Only 2 had complete wound healing, but all presented a reduction of the lesion area of approximately 22.2% and PUSH score of 9.8 to 6.6. This study found that hydrogel showed good results for the treatment of diabetic feet, reducing the area and overall PUSH score of the wounds.

Evaluation of Radiosterilized Glyercerolated Amniotic Membranes as a Substrate for Cultured Human Epithelial Cells.

Human amniotic membrane (HAM) is a biomaterial with biological properties beneficial to tissue repair, serving as a substrate for cell cultivation. Irradiation is used for tissue sterilization, but can damage the HAM structure. The objective of this paper was to construct a skin substitute, composed of human keratinocytes cultured on glycerolated HAMs, and to evaluate the influence radiation on subsequent cell culture growth. Four batches of HAMs were glycerolated, and half of them were radio-sterilzed with 25 kGy. Non-irradiated glycerolated HAM (ni-HAM) and irradiated glycerolated HAM (i-HAM) samples were then de-epithelized and analyzed using optical microscopy (Picrossirius staining), immunofluorescence and electron microscopy. Subsequently, keratinocytes were cultured on ni- and i-HAMs, and either immersed or positioned at the air-liquid interface. The basement membranes of the ni-HAM group remained intact following de-epithelialization, whereas the i-HAM group displayed no evidence or remnant presence of these membranes. Concerning the keratinocyte cultures, the ni-HAM substrate promoted the growth of multi-layered and differentiated epithelia. Keratinocytes cultured on i-HAM formed epithelium composed of three layers of stratification and discrete cell differentiation. The glycerolated HAM was compatible with cultured epithelia, demonstrating its potential as a skin substitute. Irradiation at 25 kGy caused structural damage to the amnion.

The Role of Biological Skin Substitutes in Stevens-Johnson Syndrome: Systematic Review.

Stevens-Johnson syndrome (SJS) is a disease that causes skin exfoliative lesions, characterized by fever, necrosis, and epidermal detachment. Biological skin substitutes may be considered interesting options for the treatment of this disease. This study aims at identifying in the literature the evidence on the current role of these biomaterials in the treatment of SJS and its related diseases. A systematic review with a search period between 2003 and 2017 was carried out, consulting the Lilacs, BVS, and PubMed databases. The quality of the included studies was evaluated by the Oxford Center for Evidence-Based Medicine Classification, for evaluating levels of evidence from the scientific publications. Ninety-five publications were found, and after applying inclusion and exclusion criteria, they resulted in 9 articles. In total, 20 patients with 73.87% average of body surface affected received SJS skin treatment with some biological substitutes, among them are cutaneous allograft, amnion, and xenograft. Mortality rate was 10%, and in these situations, literature indicates mortality rates ranging from 25% to 70%. Effectiveness of the use of biological dressings may be a possible explanation for this finding. Findings indicate the use of these biomaterials may favor reepithelialization, reduce water loss, decrease the chance of infection, and, consequently, improve the survival of patients with SJS and toxic epidermal necrolysis. Biological skin substitutes have characteristics that make them very promising in the topical treatment of these wounds, but their use remains very restricted in the treatment of SJS.

Effects of Hydrogel With Enriched Sodium Alginate in Wounds of Diabetic Patients.

Objective of this study was to evaluate the efficacy of the autolytic debridement promoted by hydrogel with sodium alginate enriched with fatty acids and vitamins A and E in the healing of foot wounds in diabetic patients. A clinical study was conducted at an outpatient clinic of medical specialties. The sample comprised 8 patients supervised for a 3-month period, from April to July 2017, by means of a clinical history, photographic record, planimetry, and classification of the wound severity by the Pressure Ulcer Scale for Healing (PUSH) system. Of the 8 patients supervised, 1 dropped out and 7 were followed up for 12 weeks. Only 2 had complete wound healing, but all presented a reduction of the lesion area of approximately 22.2% and PUSH score of 9.8 to 6.6. This study found that hydrogel showed good results for the treatment of diabetic feet, reducing the area and overall PUSH score of the wounds.

Estabelecimento de protocolo para armazenamento de pele autógena refrigerada / Establishment of a protocol for storage of refrigerated autologous skin

Introdução: Enxertos de pele autólogos são utilizados em tratamento de pacientes queimados. Esses enxertos podem ser armazenados e preservados, desde que o processo de armazenamento seja realizado com rígido controle de qualidade, para garantir a redução dos riscos de infecção. Métodos: Foi realizado um estudo de coorte retrospectivo na Unidade de Queimados do Hospital das Clínicas de São Paulo no período de fevereiro de 2015 a julho de 2016, em que foi estabelecido um protocolo para armazenamento de pele refrigerada com controle de coleta, preservação, embalagem e registro de todos os processos. Para garantia de qualidade, foram coletadas biópsias dos enxertos para microbiologia pré e pós-armazenamento e realizado um estudo transversal de prevalência de contaminação pré e pós-estocagem. Resultados: Os pontos críticos encontrados foram inadequação de embalagem, ausência de registros de processos, falta de coleta de biópsias para microbiologia e falhas no descarte. A maior parte das amostras estava contaminada tanto pré como pós-estocagem (84,2%). Apenas dois pacientes apresentaram microbiologia estéril no pré e contaminada no pós, porém foram encontrados germes da pele do tipo gram+. Conclusão: Foi estabelecido um método promissor de armazenamento de pele refrigerada que necessita alguns pequenos ajustes para adequação ao controle de qualidade.

Introduction: Autologous skin grafts are used for treatment of burn patients. These grafts can be stored and preserved, as long as the storage process is performed with strict quality control to reduce the risk of infection. Methods: A retrospective cohort study was conducted in the Burn Unit of the Hospital das Clínicas de São Paulo from February 2015 to July 2016. During this period, a protocol was established to store refrigerated skin, with control of collection, preservation, and packaging, and recording of all processes. To ensure quality, graft biopsies were collected for pre- and poststorage microbiology testing and a cross-sectional study for contamination was performed. Results: Critical deficiencies included inadequate packaging, lack of processing records, lack of biopsies for microbiology testing, and failure to discard specimens. Most of the samples were contaminated before and after storage (84.2%). Only two samples were sterile before storage but became contaminated after storage, with growth of Gram-positive skin bacteria. Conclusion: A promising method for the storage of refrigerated skin was established, but requires minor adjustments in quality control.

Perfil epidemiológico do doador de pele do Banco de Tecidos do Hospital das Clínicas da Universidade de São Paulo / Epidemiological characteristics of skin donors from São Paulo University Clinical Hospital / El perfil epidemiológico de los donantes de piel del Banco de tejidos del Hospital de las Clínicas de la Universidad São Paulo

OBJETIVO: Identificar o perfil epidemiológico dos doadores de pele do Banco de Tecidos do Hospital das Clínicas da Universidade de São Paulo. MÉTODO: Trata-se de um estudo documental, retrospectivo, transversal, com abordagem quantitativa de análise de prontuários de doadores de pele do Banco de Tecidos do Hospital das Clínicas de São Paulo, no período entre 2000 a 2015, para identificar as principais características epidemiológicas dos doadores de pele sendo analisadas por método estatístico descritivo. Inicialmente, foram encontrados 183 prontuários, porém 15 foram excluídos devido à falta de informações completas dos doadores. Aprovado pela Comissão de Ética para Análise de Projetos de Pesquisa do HCFMUSP (Nº CAAE 42751915.5.0000.0068). RESULTADOS: Entre os doadores pesquisados, houve predomínio do sexo masculino, com média de idade 43,3 anos, sendo a principal causa dos óbitos os acidentes cerebrais vasculares e causas externas. A retirada do tecido ocorre em geral entre 1 e 4 horas (76,1% dos casos) após a parada cardíaca. A maior parte recebeu antibioticoterapia (92,2%) e não apresenta comorbidades (81,54%). CONCLUSÃO: O doador de pele em geral é do sexo masculino, jovem e previamente hígido, sendo fundamental esta caracterização para o fortalecimento dos mecanismos de busca de doadores e aumento da oferta de tecidos no país.(AU)

OBJECTIVE: To identify the skin donors epidemiological profile from Clinical Hospital Tissue Bank. METHODS: This is a retrospective, cross-sectional documentary study with a quantitative approach to analysis medical records of skin donors from Clínicas' Hospital Tissue Bank, between 2000 and 2015, to identify the main epidemiological characteristics of skin donors, being analyzed by descriptive statistical method. Initially, 183 medical records were found, but 15 were excluded due to lack of complete donor information. Approved by the Ethics Committee for Analysis of Research Projects of HCFMUSP (CAAE No. 42751915.5.0000.0068). RESULTS: Among the donors surveyed, there was a predominance of males, with a mean age of 43.3 years, the main cause of death was cerebrovascular accidents and external causes. Tissue withdrawal usually occurs between 1 and 4 hours (76.1% of cases) after cardiac arrest. The majority received antibiotic therapy (92.2%) and was healthy (81.54%). CONCLUSION: The skin donor in general is male, young and previously healthy, and this characterization is fundamental for the strengthening of donor search mechanisms and an increase in the supply of tissues in Brazil.(AU)

Objetivo: Identificar el perfil epidemiológico de los donantes del Banco de Tejidos del Hospital das Clínicas de la Universidad de São Paulo. Método: Se realizó un estudio documental, retrospectivo, transversal, con un enfoque cuantitativo para el análises de las histórias clínicas de los donadores de piel del Banco de tejidos del Hospital das Clínicas de São Paulo, en el periodo de 2000 a 2015, para identificar las principales características epidemiológicas de los donantes de piel, siendo analisados popr método estadístico descriptivo. Inicialmente, se encontraron 183 registros, pero 15 fueron excluidos debido a la falta de información completa de los donantes. Aprobado por el Comité de Ética en Investigación (núm. CAAE 42751915.5.0000.0068). Resultados: Entre los donantes estudiados, hubo un predominio del sexo masculino, una edad media de 43,3 años, es la principal causa de muerte de los accidentes vasculares cerebrales y las causas externas. El tejido extraído es generalmente entre 1 y 4 horas (76,1% de los casos) después de un paro cardíaco. La mayoría recibió antibióticos (92,2%) y muestra comorbilidades (81,54%). Conclusión: La piel del donante es generalmente de sexo masculino, jóvenes y previamente sanos, siendo fundamental la caracterización para fortalecer los mecanimos de bísqueda de donadores y aumento de la oferta de tejidos en el país.(AU)

Oxysterols in adipose tissue-derived mesenchymal stem cell proliferation and death.

Mesenchymal stem cells (MSCs) are multipotent cells characterized by self-renewal and cellular differentiation capabilities. Oxysterols comprise a very heterogeneous group derived from cholesterol through enzymatic and non-enzymatic oxidation. Potent effects in cell death processes, including cytoxicity and apoptosis induction, were described in several cell lines. Very little is known about the effects of oxysterols in MSCs. 7-ketocholesterol (7-KC), one of the most important oxysterols, was shown to be cytotoxic to human adipose tissue-derived MSCs. Here, we describe the short-term (24h) cytotoxic effects of cholestan-3α-5ß-6α-triol, 3,5 cholestan-7-one, (3α-5ß-6α)- cholestane-3,6-diol, 7-oxocholest-5-en-3ß-yl acetate, and 5ß-6ß epoxy-cholesterol, on MSCs derived from human adipose tissue. MSCs were isolated from adipose tissue obtained from three young, healthy women. Oxysterols, with the exception of 3,5 cholestan-7-one and 7-oxocholest-5-en-3ß-yl acetate, led to a complex mode of cell death that include apoptosis, necrosis and autophagy, depending on the type of oxysterol and concentration, being cholestan-3α-5ß-6α-triol the most effective. Inhibition of proliferation was also promoted by these oxysterols, but no changes in cell cycle were observed.

Comparisons of the results of peripheral nerve defect repair with fibrin conduit and autologous nerve graft: An experimental study in rats.

INTRODUCTION: The standard treatment for nerve defects is nerve autograft. There is no conduit available that provides the same regenerative capacity of nerve autograft. This study evaluated the histological and functional recovery of nerve defects treated with fibrin conduit in comparison to the nerve autograft, in a rat model. METHOD: A sciatic nerve injury model (10-mm defect) was performed in 20 Wistar rats, nerve defect was reconstructed using a fibrin conduit (n = 10). A nerve autograft was used as control (n = 10). The walking behavior was measured by footprint analysis at 4, 8, and 12 weeks and sciatic function index was determined. After 12 weeks, histological analysis was performed to evaluate the regenerated nerve and measured axonal density. The triceps surae muscle weight was also evaluated. RESULTS: The fibrin conduit group showed less improvement in walking behavior compared to nerve autograft (-53 ± 2 vs. -36 ± 2; P < 0.001 at 12 weeks). The fibrin conduit group presented axonal density of 40.0 axons/10.995µm2 and the nerve autograft group had 67.2 axons/10.995µm2 (P < 0.001). The triceps surae muscle weight ratio of the fibrin conduit group was 41 ± 3% versus 71 ± 4% of the nerve autograft group (P < 0.001). CONCLUSION: The fibrin conduit could be used for nerve reconstruction following peripheral nerve injury in the rat model. However, the functional recovery in the fibrin conduit repair group was worse than that in nerve autograft group and the nerve repair with the fibrin conduit has less myelinated fibers when compared to the repair with nerve autograft.

Análise do atendimento clínico de portadores de úlceras crônicas em membros inferiores / Analysis of the clinical care of patients with chronic ulcers of the lower limbs

Introdução: Úlceras crônicas em membros inferiores podem apresentar diferentes etiologias, sendo as mais frequentes: venosa, arterial, traumática, infecciosa e diabética. O tratamento dessas feridas é dinâmico e depende da evolução da reparação tecidual. Esse tratamento inclui métodos clínicos e cirúrgicos, sendo o curativo o método não cirúrgico mais frequentemente utilizado. Curativos podem ser desde de coberturas inertes até veículos para atuação de princípios ativos no leito da ferida. A principal indicação de ativos está relacionada a efeitos de desbridamento e controle da população bacteriana, possibilitando o preparo desses leitos para resolução cirúrgica ou espontânea. Método: Esse estudo é observacional, longitudinal, retrospectivo, de amostra randomizada no qual pretendemos analisar o atendimento prestado aos portadores de úlceras crônicas em membros inferiores no Ambulatório de Feridas Crônicas da Divisão de Cirurgia Plástica do HCFMUSP entre 2011 e 2013. Resultados: Foram analisados prontuários de indivíduos de ambos os gêneros, idade média 60 anos, portadores de úlceras crônicas em diferentes estágios evolutivos. Foram pesquisadas comorbidades, doença de base, tamanho da lesão, tratamentos utilizados e evolução das feridas. Notouse predomínio das causas vasculares (69,2%) como doenças de base. Todos os pacientes foram primeiramente tratados com curativos contendo princípios ativos, para preparo do leito das feridas. Desses, 84% foram encaminhados para resolução cirúrgica das feridas. Houve fechamento espontâneo em 1,5% dos casos. Os demais pacientes (14,5%) apresentaram piora das lesões com tratamento tópico, necessitando outras formas de preparo desse leito. Conclusão: Agentes tópicos podem ser uma forma ambulatorial/domiciliar efetiva de preparo do leito de úlceras crônicas para resolução cirúrgica.

Introduction: Chronic ulcers of the lower limbs may have different etiologies, with the most frequent being venous, arterial, traumatic, infectious, and diabetic. The treatment of these wounds is dynamic and depends on the evolution of tissue repair. This treatment includes clinical and surgical methods, and dressings are the most frequently used. Dressings can range from inert covers to vehicles for actuation of active substances in the wound bed. The main indication for these substances is related to the effects of debridement and control of the bacterial population, enabling the preparation of wound beds for surgical or spontaneous resolution. Method: This study is an observational, cross-sectional, retrospective study, with random sampling, aimed at assessing the care provided to patients with chronic ulcers of the lower limbs in the Outpatient Clinic for chronic wounds of the Plastic Surgery Division, HCFMUSP, between 2011 and 2013. Results: The clinical charts of 481 patients of both sexes, with a mean age 60 years, were analyzed; all had chronic ulcers in different stages of evolution. Comorbidities, underlying disease, size of the lesion, treatments, and evolution of wounds were evaluated. A predominance of vascular disease (69.2%) was the underlying cause. All patients were initially treated with dressings containing active agents, for preparation of the wound bed. Of these, 84% were referred for surgical management of wounds. There was no spontaneous closure in 1.5% of cases. The remaining patients (14.5%) showed worsening of lesions with topical treatment, and required other forms of preparation of the wound bed. Conclusion: Topical agents may be an effective outpatient/home method to prepare the wound bed of chronic ulcers for surgical management.

Estudo comparativo entre pentoxifilina e triancinolona no tratamento das cicatrizes hipertróficas pós-queimadura quanto à remodelação do colágeno e das fibras do sistema elástico / Comparative study of pentoxifylline and triamcinolone in the treatment of post-burn hypertrophic scars on the collagen remodeling and elastic system fibers

Introdução: O tratamento das sequelas cicatriciais permanece um desafio na prática diária. Corticosteroides injetáveis são amplamente utilizados no combate a queloides e cicatrizes hipertróficas, mas substâncias como a pentoxifilina (PTF) também têm demonstrado eficácia clínica na modulação dessas cicatrizes. Objetivos: No presente estudo, propusemos a comparação dos efeitos da PTF e do corticosteroide triancinolona nas cicatrizes hipertróficas de pacientes vítimas de queimaduras por meio de análise histológica da organização das fibras que contêm colágeno e das fibras do sistema elástico. Métodos: Foram estudadas amostras de pele cicatricial de 10 pacientes, entre 20 e 40 anos, com história de queimaduras em tronco, com até 24 meses de evolução, não tratadas cirurgicamente. Cada paciente teve duas áreas cicatriciais tratadas, uma com Hexacetonido de Triancinolona 20 mg/ml e outra com Pentoxifilina 1 mg/ ml; tendo sido realizadas três aplicações intracicatriciais com intervalos mensais. Uma biópsia de cada área tratada foi colhida após 30 dias de cada aplicação. Resultados: Os resultados clínicos foram evidentes e semelhantes para as duas drogas: diminuição da espessura, do prurido, da hiperemia e da consistência da cicatriz. Não se observaram diferenças arquiteturais no tecido conjuntivo subepidérmico quando comparadas a cicatriz original com as cicatrizes após cada tipo de tratamento (grandes feixes de fibras colágenas em todas as direções, com ausência de fibras do sistema elástico). Estudos subsequentes envolvendo a análise da espessura total da cicatriz e o grau de vascularização/ inflamação presentes se fazem necessários na investigação da justificativa da eficácia clínica dos tratamentos. Conclusão: Concluímos que a PTF teve uma resposta clínica e morfológica similar à triancinolona nos casos tratados.

Introduction: The treatment of scarring sequelae remains challenge in daily practice. Injecting corticosteroids are widely used to combat keloids and hypertrophic scars, but substances such as pentoxifylline (PTF) have also demonstrated clinical efficacy in modulating these scars. Objectives: This study set out to compare the effects of TFP and corticosteroid triamcinolone in hypertrophic scars of burn victims by histological analysis of the organization of the fibers containing collagen and elastic system fibers. Methods: Scar skin samples from 10patients were studied between 20 and 40 years, with a history of burns on the trunk, up to 24 months of evolution, not surgically treated. Each patient had two treated scar areas, one with triamcinolone hexacetonide 20 mg/ml and the other with pentoxifylline 1 mg/ml; having been held three intracicatriciais applications at monthly intervals. A biopsy of each treated area was harvested after 30 days of each application. Results: The clinical results were evident and similar for the two drugs: thinning, itching, hyperemia and scar consistency. There were no differences in architectural subepidermal connective tissue when compared with the original scar scars after each treatment (large bundles of collagen fibers in all directions with no elastic system fibers). Subsequent studies involving the analysis of the total thickness of the scar and the extent of vascularization/inflammation gifts are needed to investigate the reasons of clinical efficacy of treatments. Conclusion: We conclude that TFP had a clinical and morphological response similar to triamcinolone in treated cases.

Short-term effects of 7-ketocholesterol on human adipose tissue mesenchymal stem cells in vitro.

Oxysterols comprise a very heterogeneous group derived from cholesterol through enzymatic and non-enzymatic oxidation. Among them, 7-ketocholesterol (7-KC) is one of the most important. It has potent effects in cell death processes, including cytoxicity and apoptosis induction. Mesenchymal stem cells (MSCs) are multipotent cells characterized by self-renewal and cellular differentiation capabilities. Very little is known about the effects of oxysterols in MSCs. Here, we describe the short-term cytotoxic effect of 7-ketocholesterol on MSCs derived from human adipose tissue. MSCs were isolated from adipose tissue obtained from two young, healthy women. After 24 h incubation with 7-KC, mitochondrial hyperpolarization was observed, followed by a slight increase in the level of apoptosis and changes in actin organization. Finally, the IC50 of 7-KC was higher in these cells than has been observed or described in other normal or cancer cell lines.

Construção de substituto da pele composto por matriz de colágeno porcino povoada por fibroblastos dérmicos e queratinócitos humanos: avaliação histológica / Construction of a skin substitute composed of porcine collagen matrix populated with human dermal fibroblasts and keratinocytes: histological evaluation", "_e": "n

INTRODUÇÃO: O uso de enxertos autólogos é limitado pela extensão da área doadora e pelo estado clínico dos pacientes, no caso de lesões extensas. Alotransplantes coletados de cadáveres ou voluntários são rejeitados após uma ou duas semanas, servindo apenas como cobertura temporária para essas lesões. O tratamento de grandes lesões cutâneas com tegumento autólogo reconstruído constitui alternativa atraente, já que, a partir de um pequeno fragmento de pele do paciente, pode-se obter culturas de células que se multiplicam rapidamente e podem ser criopreservadas, permitindo, assim, sua utilização em novos tratamentos por tempo indeterminado. Este estudo pretendeu avaliar o comportamento histológico de queratinócitos e fibroblastos humanos cultivados sobre uma matriz de colágeno porcino derivada da submucosa intestinal. MÉTODO: Células da epiderme e derme humana foram cultivadas separadamente e semeadas sobre matriz de colágeno porcino, onde permaneceram em ambiente controlado por 21 dias, antes de serem submetidas a análise histológica. RESULTADOS: Observou-se que os fibroblastos invadem e colonizam a matriz de colágeno, enquanto os queratinócitos se organizam de forma laminar e estratificada sobre a superfície em que foram semeados. CONCLUSÕES: A utilização da matriz de colágeno porcino como carreador de células da pele humana é possível e a organização dessas células se assemelha à arquitetura da pele humana.

BACKGROUND: In the case of extensive lesions, the use of autologous grafts is limited by the extent of the donor area and the clinical condition of patients. Allografts collected from cadavers or volunteers are usually rejected after 1 to 2 weeks, thus serving only as temporary cover for these lesions. Treating major cutaneous lesions with reconstructed autologous skin is an attractive alternative, because it is possible to obtain cultures of cells that multiply rapidly and can be cryopreserved from a small fragment of the patient's skin, thereby facilitating its indefinite use in new treatments. This study evaluated the histological behavior of cultured human keratinocytes and fibroblasts on a collagen matrix derived from porcine small intestinal submucosa. METHODS: Cells from human epidermis and dermis were grown separately and seeded on porcine collagen matrix, which was maintained in a controlled environment for 21 days before being subjected to histological analysis. RESULTS: Fibroblasts invaded and colonized the collagen matrix, whereas keratinocytes were organized in laminated and stratified layers on the surface on which they were seeded. CONCLUSIONS: The use of porcine collagen matrix as a support for human skin cells is feasible, and the organization of these cells resembles the architecture of human skin.

Sistematização de curativos para o tratamento clínico das feridas / Systematization of dressing for clinical treatment of wounds", "_e": "n

O tratamento das feridas cutâneas inclui métodos clínicos e cirúrgicos, sendo o curativo um dos tratamentos clínicos mais frequentemente utilizados. Um vasto arsenal terapêutico composto por curativos passivos ou com princípios ativos é capaz de auxiliar na reparação do tegumento em diversas situações. Curativos visam a melhorar as condições do leito da ferida, podendo ser, em algumas ocasiões, o próprio tratamento definitivo, mas em muitas situações constituem apenas uma etapa intermediária para o tratamento cirúrgico. Curativos inteligentes e biológicos são hoje mais bem classificados como substitutos cutâneos e não serão considerados neste artigo. A escolha do curativo a ser utilizado deve ser baseada no conhecimento das bases fisiopatológicas da cicatrização e da reparação tecidual, sem nunca esquecer o quadro sistêmico do paciente.

The treatment of cutaneous wounds includes both medical and surgical methods; dressing is one of the most commonly used clinical treatments. An extensive therapeutic toolkit comprising passive dressings or dressings with active principles can help repair wounds in various situations. Dressings are used to improve the conditions of the wound bed and may occasionally be considered the definitive treatment, whereas in some cases, they may be considered an intermediate step to surgical treatment. Intelligent and biological wound dressings are currently classified as dermal substitutes and will not be discussed in this article. Dressings should be selected on the basis of knowledge of the pathophysiology of wound healing and tissue repair while keeping the systemic problems of the patient in mind.

Cultivo de células-tronco derivadas de tecido adiposo: uma análise crítica / Culture of adipose-derived stem cells:a critical analysis

Introdução: células-tronco derivadas de tecido adiposo (ADSCs) são pluripotentes, tem coleta poucoinvasiva e apresentam alto rendimento celular em cultura, além do fato de o número de lipoaspirações, a principalfonte destas células, estar aumentando ao redor do mundo. A Cirurgia Plástica tem encontrado benefícios de seu usoem diversas áreas, o que ressalta a importância de se analisar criticamente os protocolos de cultivo atualmenteutilizados. Objetivo: descrever detalhadamente o protocolo de cultivo adotado pelo nosso laboratório para podermos realizar uma análise crítica sobre o cultivo de ADSCs. Método: excedentes de tecido adiposo foram doados ao laboratório com finalidade de pesquisa por cinco pacientes de ambos os sexos e faixa etária de 20 a 45 anos. De cada paciente, 30 mL do material gorduroso foram coletados em frasco estéril e transportados ao laboratório de cultura celular, onde foram submetidos, por 45 min, à digestão enzimática por 30 mg de colagenase tipo IA diluída em30mL de meio de Dulbecco Modificado por Eagle (DMEM) e posteriormente centrifugados para que se isolem ascélulas da matriz extracelular. A amplificação das culturas foi feita com solução de tripsina 0,05% + EDTA 0,02%. Para congelamento, utilizou-se meio de congelamento composto por 60% DMEM, 10% dimetilsulfóxido (DMSO) e 30% SBF. Resultados: o protocolo apresentado para o cultivo de ADSCs demonstrou um cultivo celularsatisfatório, completando três passagens de cultura em todas as amostras, com posterior congelamento edescongelamento. Conclusões: a literatura científica nos indica que a colagenase possibilita extração e isolamento de ADSCs, devendo-se atentar ao tipo utilizado. Quanto ao congelamento, dado a citotoxicidade do DMSO, novos protocolos com grandes amostras substituindo este crioprotetor por trealose devem ser testados, visto que este último produziu bons resultados e não apresentou efeitostóxicos nos estudos já publicados.

Introduction: adipose-derived stem cells (ADSCs) are pluripotent and present little invasive collectionand high cell yield in culture, besides the fact that the number of liposuctions, the main source of these cells, is increasing around the world. Plastic Surgery has found the benefits of the use of those cells in several areas, which increases the importance of critically analyzing the protocolscurrently used for cell cultivation. Objective: to describe in detail the cultivation protocol adopted by our laboratory in order to perform a critical analysis of the culture of ADSCs. Method: surpluses of adipose tissue were donated to the laboratory with research purposes for 5 patients of bothgenders and age between 20 and 45 years. From each patient, 30 mL of fatty material was collected in a sterile flask and transported to the laboratory for cell culture, where they were subjected during 45 min to enzymatic digestion by 30 mg of type IA collagenase diluted in 30mL of Dulbecco’s Modified Eagle Medium (DMEM) and thencentrifuged to isolate the cells from the extracellular matrix. The amplification of the cultures was made with 0.05%trypsin solution + EDTA 0.02%. For cryopreservation was used freezing media consisting of 60% DMEM, 10%dimethylsulfoxide (DMSO) and 30% FBS. Results: The above protocol for growing ADSCs obtained a satisfactorycell culture, completing three cell passages in all samples, with subsequent freezing and thawing. Conclusions: The scientific literature indicates that collagenase get the bestresults in the extraction and isolation of ADSCs, paying attention to the type used. Regarding freezing, given thecytotoxicity of DMSO, new protocols with large samples replacing this cryoprotectant by trehalose should be tested, since the latter has produced good results and showed no toxic effects in published studies.

Substitutos cutâneos: conceitos atuais e proposta de classificação / Skin substitutes: current concepts and a new classification system

Feridas complexas podem resultar na perda completa do revestimento cutâneo. A solução consagrada pela cirurgia plástica é a enxertia de pele autógena, porém há casos em que ocorre escassez de área doadora cutânea, um problema ainda não totalmente solucionado. Assim, atualmente há muito interesse por materiais sintéticos ou biológicos que possam ser utilizados como substitutos cutâneos. O objetivo deste estudo foi introduzir uma forma mais didática de agrupar diferentes tipos de substitutos cutâneos. Acreditamos que esses produtos podem ser classificados de forma mais abrangente se forem divididos segundo três critérios: camada substituída da pele, subdivididos em epidérmicos (E), dérmicos (D) e compostos dermoepidérmicos (C); duração no leito da ferida, subdivididos em temporários (T) e permanentes (P); e origem do material constituinte, subdivididos em biológicos (b), biossintéticos (bs) e sintéticos (s).

Complex wounds are characterized by complete loss of cutaneous cover. The most common plastic surgery technique is the autogenous skin graft; however, the amount of material available from donor areas is often limited. The development of synthetic or biological products as skin substitutes is therefore an area of interest. The present study aimed to classify the different types of skin substitutes available based on three criteria: the skin layer to be replaced, which can be categorized into epidermal (E), dermal (D), and dermal-epidermal composites (C); the durability in the wound bed, which can be temporary (T) or permanent (P); and the origin of the material, subdivided into biological (b), biosynthetic (bs), and synthetic (s).

Clinical trial comparing 3 different wound dressings for the management of partial-thickness skin graft donor sites.

PURPOSE: A review of the literature reveals a lack of consensus regarding local management of skin graft donor sites. This study was undertaken to determine the effects of 3 different dressings on healing of donor sites and patient discomfort related to donor sites. DESIGN: This study is a comparison cohort study. SUBJECTS AND SETTING: We recruited 34 burn patients scheduled for partial-thickness skin grafts; their mean age was 36 ± 18 years (mean ± SD, range 20 to 54 years), and 63% were male. All subjects were managed at Burn Unity Care, located in Sao Paulo, Brazil. MATERIALS AND METHODS: Subjects were randomly allocated to 1 of 3 groups: (1) a study group (group A) whose donor sites were dressed with a bovine collagen calcium-alginate dressing covered with transparent polyurethane film; (2) a second intervention group (group B), whose donor sites were dressed only with transparent polyurethane film dressing; and (3) a control group whose donor sites were dressed with rayon soaked in 0.9% saline (group C). Two independent observers assessed donor site wounds for epithelialization, scabbing, quantity and characteristics of exudate, and complications. Pain was measured using the visual analog scale, the brief pain inventory, and Index of Pain Management. RESULTS: Subjects managed with the bovine collagen calcium-alginate dressing covered with transparent polyurethane film (group A) achieved the greatest epithelialization (6.3 vs 8.2 for thin film dressing only P < .02 and 6.3 vs 11.7 days for control group P < .01). Patients managed with the bovine collagen calcium-alginate dressing covered with transparent polyurethane film also reported less pain that subjects allocated to the control (group C) or thin film only group (group B), (P < .05). Ninety percent of subjects allocated to the calcium alginate covered with thin film dressing reported mild pain intensity on the Visual Analog Scale, 85% of did not report pain localized to the donor site on the brief pain inventory, and scores on the Index of Pain Management ranged from 23 to 11. CONCLUSION: Study findings suggest that use of a collagen calcium-alginate dressing with a transparent film covering reduces the time for complete epithelialization and may reduce pain related to skin graft donor sites.

Role of keratinocytes in wound contraction: an impact assessment using a model of collagen matrix populated with fibroblasts / Papel do queratinócito na contração da ferida: avaliação de impacto usando um modelo de matriz de colágeno povoada por fibroblastos

BACKGROUND: The possible participation of keratinocytes in wound remodeling has been widely studied. This study investigated the impact of keratinocytes in wound contraction. METHODS: Murine type I collagen gels populated by human fibroblasts and seeded with human keratinocytes on the surface to form a dermo-epidermal equivalent were used as the study group. Collagen gels populated by only fibroblasts were used as the control group. The criteria for the preparation and storage of gels were similar for both groups. RESULTS: An evident and statistically significant increase in gel contraction was observed in samples populated by keratinocytes compared to the control group. CONCLUSIONS: These results suggest that keratinocytes not only modulate fibroblast proliferation but also play an active role in wound contraction per se. Further research on the mechanisms involved in the communication pathways between cells and between cells and the matrix shall be assessed from the perspective of keratinocyte participation in wound healing and pathologic scarring.

INTRODUÇÃO: A eventual participação de queratinócitos na remodelagem da ferida tem sido estudada há muito tempo. Este trabalho investigou o impacto dos queratinócitos na contração da ferida. MÉTODO: Foi utilizado gel de colágeno tipo I murino povoado por fibroblastos humanos com queratinócitos humanos semeado na superfície (grupo estudo), formando um equivalente dermoepidérmico. Géis de colágeno povoado apenas por fibroblastos foram utilizados como grupo controle. Os critérios de confecção e armazenagem dos géis foram iguais para ambos os grupos. RESULTADOS: Houve aumento evidente e estatisticamente significante na contração de gel das amostras povoadas por queratinócitos, em comparação ao grupo controle. CONCLUSÕES: Esses resultados sugerem que os queratinócitos não só podem modular a proliferação de fibroblastos, mas também, por si só, desempenhar papel ativo na contração da ferida. Novas investigações sobre mecanismos envolvidos nas vias de comunicação entre células e entre célula e matriz devem ser avaliadas sob o ponto de vista de participação dos queratinócitos na cicatrização de feridas e formação de cicatrizes patológicas.

Replacement of fetal calf serum by human serum as supplementation for human fibroblast culture / Substituição de soro bovino fetal por soro humano como suplemento para cultura de fibroblastos humanos

INTRODUCTION: Fetal calf serum (FCS) is commonly used as a supplement in the culture medium for fibroblast cells. This supplementation is far from ideal as sample quality varies from batch to batch and the composition of FCS is not completely known. In addition, FCS may be contaminated with viruses and/or prions and may also cause adverse immunologic responses in humans. Due to these facts, a worldwide effort is being made to find alternatives for xenobiotic elements in cell cultures. Human serum could be a safer alternative, especially for clinical application. METHODS: We investigated human serum as a substitute for FCS in human fibroblast culture. Fresh human serum was obtained from 10 healthy volunteers. Fibroblasts were cultivated in multiwell plates containing either Dulbecco's modified Eagle's medium (DMEM) plus 10 percent FCS (D10) or DMEM plus 10 percent human serum (D10H). Cell counts were obtained between 24 and 264 hours of cultivation; results were expressed as the mean number of cells ± standard error of the mean to create cell proliferation curves. RESULTS: There was no statistical difference in fibroblast proliferation between the two groups. Human serum supported human fibroblast growth and proliferation, suggesting that it may be a potential substitute for FCS in human cell culture. Cells cultivated with human serum presented a different morphology, appearing smaller and more rounded as compared to cells cultivated in D10. CONCLUSIONS: These results demonstrate that human serum can be substituted for FCS in human fibroblasts culture and that fibroblasts cultivated in the presence of human serum have a morphology that is similar to in vivo fibroblasts.

INTRODUÇÃO: Soro bovino fetal (SBF) é comumente usado como suplemento no meio de cultura para cultivar fibroblastos. Essa forma de suplementação, porém, não é ideal, pois a qualidade das amostras de SBF é variada e sua composição não é completamente conhecida. Além disso, o SBF pode apresentar contaminação por vírus e príons ou causar complicações imunológicas. Assim, a comunidade científica tem buscado alternativas ao uso de elementos xenobióticos em cultura celular. O soro humano pode ser uma dessas alternativas, principalmente para aplicação clínica. MÉTODO: Soro humano, obtido de sangue de 10 voluntários saudáveis submetidos a avaliação sorológica prévia, foi testado como substituto do SBF em cultura de fibroblastos humanos. As células foram cultivadas em placas multipoços, contendo Dulbecco's Modified Eagle's Medium (DMEM) mais 10 por cento de SBF (D10) ou DMEM mais 10 por cento de sroro humano (D10H). Entre 24 e 264 horas de exposição aos meios testados, as células foram contadas e os resultados foram expressos em média ± erro padrão da média, para obtenção de curvas de proliferação celular. RESULTADOS: Não houve diferença estatística entre os grupos de proliferação. Fibroblastos na presença de soro humano aparentavam ser menores e mais arredondados em comparação àqueles mantidos em D10. CONCLUSÕES: Os resultados permitem inferir que o soro humano pode substituir o SBF em cultura de fibroblastos e que fibroblastos cultivados em meio suplementado por soro humano apresentam morfologia mais semelhante àqueles in vivo.