RESUMO Introdução: Embora o tratamento do câncer de cabeça e pescoço tenha evoluído nos últimos anos, ainda é um desafio preservar e restaurar a simetria facial após o mesmo, comprometendo significativamente a qualidade de vida dos pacientes. Em cirurgia reconstrutiva, o enxerto de gordura autóloga vem sendo usado para tratar defeitos de volume e de contorno. O objetivo é revisar os efeitos do enxerto de gordura em pacientes com radiodermite, devido ao tratamento radioterápico de câncer de cabeça e pescoço. Métodos: Revisão de literatura nas bases PubMed e SciELO, buscando estudos publicados nos últimos 20 anos nas línguas portuguesa, inglesa ou espanhola. Foram utilizados os descritores “adipose tissue”, “transplantation”, “neoplasms”, “head and neck neoplasms” e “radiotherapy”; e os termos não descritores “fat grafting”, “fat transplantation” e “fat graft”. Resultados: De 212 artigos encontrados, apenas 7 estavam em conformidade aos critérios de elegibilidade. Conclusão: A revisão sugere que a lipoenxertia em reconstrução oncológica de cabeça e pescoço é um tratamento seguro e permite otimizar os resultados estéticos e funcionais. Faltam estudos prospectivos para melhor definição de técnicas e de resultados.

ABSTRACT Introduction: Although treatment of head and neck cancer has evolved in recent years, it is still a challenge to preserve and restore facial symmetry afterward; this fact significantly compromises the quality of life of patients. In reconstructive surgery, autologous fat grafting has been used to treat volume and contour defects. The objective is to review the effects of fat grafting in patients with radiodermatitis due to head and neck cancer radiotherapy treatment. Methods: Literature review in PubMed and SciELO databases, looking for studies published in the last 20 years in Portuguese, English, or Spanish. The descriptors “adipose tissue”, “transplantation”, “neoplasms”, “head and neck neoplasms” and “radiotherapy” and the non-descriptive terms “fat grafting”, “fat transplantation” and “fat graft” were used. Results: Of 212 articles found, only seven complied with the eligibility criteria. Conclusion: The review suggests that fat grafting in head and neck cancer reconstruction is a safe treatment and optimizes aesthetic and functional results. Prospective studies are lacking for a better definition of techniques and results.

Abstract Purpose: To assess the action of vitamin C on the expression of 84 oxidative stress related-genes in cultured skin fibroblasts from burn patients. Methods: Skin samples were obtained from ten burn patients. Human primary fibroblasts were isolated and cultured to be distributed into 2 groups: TF (n = 10, fibroblasts treated with vitamin C) and UF (n = 10, untreated fibroblasts). Gene expression analysis using quantitative polymerase chain reaction array was performed for comparisons between groups. Results: The comparison revealed 10 upregulated genes as follows: arachidonate 12-lipoxygenase (ALOX12), 24-dehydrocholesterol reductase (DHCR24), dual oxidase 1 (DUOX1), glutathione peroxidase 2 (GPX2), glutathione peroxidase 5 (GPX5), microsomal glutathione S-transferase 3 (MGST3), peroxiredoxin 4 (PRDX4), phosphatidylinositol-3,4,5-trisphosphate dependent Rac exchange factor 1 (P-REX1), prostaglandin-endoperoxide synthase 1 (PTGS1), and ring finger protein 7 (RNF7). Conclusion: Cultured fibroblasts obtained from burn patients and treated with vitamin C resulted in 10 differentially expressed genes, all overexpressed, with DUOX1, GPX5, GPX2 and PTGS1 being of most interest.

Abstract Purpose: To investigate the use Aldefluor® and N, N - Dimethylaminobenzaldehyde (DEAB) to design a protocol to sort keratinocyte stem cells from cultured keratinocytes from burned patients. Methods: Activated Aldefluor® aliquots were prepared and maintained at temperature between 2 to 8°C, or stored at -20°C. Next, the cells were collected following the standard protocol of sample preparation. Results: Best results were obtained with Aldefluor® 1.5µl and DEAB 15 µl for 1 x 106 cells, incubated at 37°C for 15 minutes. Flow cytometer range for keratinocyte stem cells separation was evaluated. There were 14.8% of stem cells separated in one sample of keratinocyte culture used to pattern the protocol. After being defined the ideal concentration, the same test pattern was performed in other keratinocyte samples. We observed a final mean of 10.8%. Conclusion: Aldefluor® has been shown as a favorable marking of epidermal keratinocyte stem cells for subsequent separation on a flow cytometer, with detection of 10.8% of epidermal keratinocyte stem cells, in this protocol.

Objetivo: Evaluar la Calidad de Vida (CV) y la capacidad funcional en pacientes con Enfermedad Arterial Periférica ( EAP) y Úlceras Arteriales (UA). Metodología: Estudio clínico descriptivo y transversal, realizado en el sur de Minas Gerais, Brasil. Sesenta pacientes adultos asignados en tres grupos -grupo EAP con UA, grupo EAP sin UA y grupo de control sin EAP y UA- fueron entrevistados, para lo cual se hizo uso de las escalas Health Assessment Questionnaire (HAQ) y el Short Form-36 Health Survey (SF-36) que evalúan la capacidad física y la calidad de vida, respectivamente. Para las comparaciones entre los grupos, se utilizó la prueba de Kruskal-Wallis. Resultados: Cuarenta y dos participantes (70%) son mujeres y la edad media fue de 63 años. Los pacientes con EAP, con o sin UA, presentaron limitación funcional significativamente mayor que los pacientes sin PAD (p < 0,01) en componentes relacionados con el andar, el uso de utensilios domésticos, la higiene y el alcance/agarre de objetos. Los pacientes con PAD, con o sin lesiones, tenían puntuaciones significativamente más bajas de SF-36 -CV inferior- en las dimensiones Función física, Rol físico, Dolor corporal, Vitalidad, Función social y Rol emocional en comparación con los pacientes del grupo control (p < 0,001). También se observaron diferencias significativas entre el grupo control y el grupo con EAP y au en las dimensiones Salud general y Salud mental. Conclusión: Los pacientes con EAP, con o sin úlcera, mostraron dificultad para realizar algunas actividades cotidianas, así como baja calidad de vida en comparación con los pacientes sin la enfermedad.

Objective: To assess Quality of Life (QOL) and functional ability in patients with Peripheral Arterial Disease (PAD) and Arterial Ulcers (AUS). Methodology: Descriptive and cross-sectional clinical study, conducted in Southern Minas Gerais, Brazil. Sixty adult patients allocated in three groups -group PAD with aus, group PAD without aus, and control group, without both PAD and aus- were interviewed using the scales Health Assessment Questionnaire (HAQ) and Short Form-36 Health Survey (SF-36), which assess physical disability and quality of live, respectively. The Krus-kal-Wallis test was used in order to compare the groups. Results: Forty-two participants (70%) are women, and the mean age was 63 years old. Patients with PAD, with or without aus, had significantly greater disability than patients without PAD (p < 0.01) on components related to walking, use of household utensils, hygiene, and reach/grip of objects. Patients with PAD, with or without lesions, reported significantly lower SF-36 scores -lower QOL- on Physical functioning, Role physical, Bodily pain, Vitality, Social functioning, and Role emotional domains, in comparison to patients from the control group (p < 0.001). Significant differences were also observed between the control group and the group with PAD and aus on the General health and Mental health domains. Conclusion: Patients with PAD, with or without AUS, reported difficulty in performing some activities of daily living and impaired QOL in comparison to patients without the disease.

Resumo Objetivo: Avaliar a Qualidade de Vida (QV) e a capacidade funcional em pacientes com Doença Arterial Periférica (DAP) e Úlceras Arteriais (UA). Metodologia: Estudo clínico descritivo e transversal, realizado no Sul de Minas Gerais, Brasil. Sessenta pacientes adultos alocados em três grupos -grupo DAP com UA, grupo DAP sem UA e grupo controle, sem DAP e sem UA- foram entrevistados, utilizando-se os questionários Health Assessment Questionnaire (HAQ) e o Short Form-36 Health Survey (SF-36) para medir a capacidade funcional e a QV, respectivamente. Para a comparação entre os grupos, foi utilizado o teste de Kruskal-Wallis. Resultados: Quarenta e dois participantes (70%) são mulheres e a idade média foi de 63 anos. Os pacientes com DAP, com ou sem UA, apresentaram incapacidade funcional significativamente maior que pacientes sem DAP (p < 0,01) nos componentes relacionados a deambulação, uso de utensílios domésticos, higiene e apreensão/sustentação de objetos. Os pacientes com DAP, com ou sem lesões, apresentaram SF-36 escores significativamente mais baixos -menor QV- nos domínios Capacidade funcional, Aspectos físicos, Dor corporal, Vitalidade, Aspectos sociais e Aspecto emocional do SF-36 em relação ao grupo controle (p < 0,001). Também foram observadas diferenças significativas (p < 0,001) entre o grupo controle e o grupo com DAP e UA nos domínios Estado geral de saúde e Saúde mental. Conclusão: Os pacientes com DAP, com ou sem úlcera, apresentaram redução na capacidade de realização de algumas atividades cotidianas, além de baixos níveis de QV quando comparados aos pacientes sem a doença.

ABSTRACT PURPOSE: To evaluate the effect of keratinocyte growth factor (KGF) treatment on the expression of wound-healing-related genes in cultured keratinocytes from burn patients. METHODS: Keratinocytes were cultured and divided into 4 groups (n=4 in each group): TKB (KGF-treated keratinocytes from burn patients), UKB (untreated keratinocytes from burn patients), TKC (KGF-treated keratinocytes from controls), and UKC (untreated keratinocytes from controls). Gene expression analysis using quantitative polymerase chain reaction (qPCR) array was performed to compare (1) TKC versus UKC, (2) UKB versus UKC, (3) TKB versus UKC, (4) TKB versus UKB, (5) TKB versus TKC, and (6) UKB versus TKC. RESULTS: Comparison 1 showed one down-regulated and one up-regulated gene; comparisons 2 and 3 resulted in the same five down-regulated genes; comparison 4 had no significant difference in relative gene expression; comparison 5 showed 26 down-regulated and 7 up-regulated genes; and comparison 6 showed 25 down-regulated and 11 up-regulated genes. CONCLUSION: There was no differential expression of wound-healing-related genes in cultured primary keratinocytes from burn patients treated with keratinocyte growth factor.

ABSTRACT Introduction: The transverse rectus abdominis myocutaneous flap is a well stablished surgery for breast reconstruction. Its main disadvantage is a large defect in its donor site, leading to weakness in the abdominal wall. Many surgical tactics has been described to address the donor site defect, including direct closure of the muscular aponeurotic and use of polypropylene mesh. The purpose of this study was to assess abdominal hernia and bulge after TRAM flap breast reconstruction using two layers of polypropylene mesh in the donor site defect. Methods: Prospective study in 24 patients who had unilateral TRAM flap breast reconstruction with two layers of polypropylene mesh to close the donor site defect. The patients were evaluated in 3.6 and 12 months of postoperative time for incidence of abdominal hernia and bulge. Results: It was observed one case of abdominal bulge (4.16%) and no case of abdominal hernia. There were no cases of mesh infection or extrusion. Conclusion: The use of two layers of polypropylene mesh in the donor site defect of TRAM flap breast reconstruction resulted in low incidence of abdominal hernia and bulge.

RESUMO Introdução: O retalho transverso do músculo reto abdominal (TRAM) é um método bem estabelecido para reconstrução mamária. Apresenta como principal desvantagem o grande defeito da área doadora, podendo causar fraqueza da parede abdominal, hérnias e abaulamentos. Diversas táticas de fechamento do defeito da área doadora foram descritas, incluindo fechamento direto da aponeurose do músculo reto abdominal e uso de telas de polipropileno. O objetivo deste estudo foi avaliar os índices de abaulamento e hérnia no pós-operatório de TRAM utilizando a tática cirúrgica de fechamento da área doadora com duas telas de polipropileno. Métodos: Estudo prospectivo em 24 pacientes submetidos à reconstrução mamária unilateral com TRAM monopediculado ou bipediculado e reparo da área doadora com duas telas de polipropileno. Os pacientes foram avaliados quanto à incidência de hérnia e abaulamento no pósoperatório de 3, 6 e 12 meses. Resultados: Ocorreu um caso (4,16%) de abaulamento abdominal e nenhum caso de hérnia. Não houve casos de infecção ou extrusão da tela. Conclusão: O uso de duas telas de polipropileno na área doadora do TRAM propiciou baixa incidência de hérnias e abaulamentos abdominais.

ABSTRACT Introduction: Despite great advances in treatment, burned skin infection remains a major challenge. The aim of this study is to evaluate the microbiological aspects of the first year’s operation of a Burn Unit in a University Hospital. Methods: Retrospective study. Microbiological data were collected and analyzed from patients admitted to the Burn Unit of São Paulo Hospital, a University Hospital of the Paulista Medical School (EPM) of the Federal University of São Paulo (UNIFESP) from June 2009 to July 2010. Results: The average length of stay was 13.8 days with a mortality rate of 5.9%, and median of TBSA was 10.3%. Evaluated 159 cultures from 101 patients. Blood cultures were the most requested (41%). It was also accessed 245 surveillance cultures collected from 75 patients. The microbiological analysis revealed a total positivity rate of 34,5%. The most prevalent agents were Coagulase-negative Staphylococcus - CoNS - (33%), Pseudomonas aeruginosa (24%), Acinetobacter spp. (22%) and Klebsiella pneumoniae (5%). Conclusion: The microbiological evaluation of the first year’s activity of EPM/UNIFESP Burn Care Unit revealed that, although the most prevalent agent was CoNS, Gram negative bacilli are still very prevalent, such as Pseudomonas aeruginosa and Acinetobacter baumannii. Despite the short time of operation, was observed large number of multiresistant microorganisms which can be explained by long exposure to antimicrobials and high transfer rate from other hospitals.

RESUMO Introdução: Apesar dos grandes avanços em seu tratamento, infecção de pele com queimadura continua a ser um grande desafio. O objetivo deste estudo é avaliar os aspectos microbiológicos do primeiro ano de funcionamento de uma unidade de queimadura em um Hospital Universitário. Métodos: Estudo retrospectivo. Dados microbiológicos foram coletados e analisados a partir de pacientes internados na Unidade de Queimadura (UTQ) do Hospital São Paulo, Hospital Universitário da Escola Paulista de Medicina (EPM) da Universidade Federal de São Paulo (UNIFESP), entre junho de 2009 e julho de 2010. Resultados: O tempo médio de permanência hospitalar foi de 13,8 dias, com uma taxa de mortalidade de 5,9%. A média da superfície corpórea queimada foi de 10,3%. Avaliou-se 159 culturas de 101 pacientes. Culturas de sangue foram as mais solicitadas (41%). Também foram acessadas 245 culturas de vigilância, coletadas de 75 pacientes. A análise microbiológica revelou um índice de positividade total de 34,5%. Os agentes mais prevalentes foram Staphylococcus coagulase-negativo - CoNS - (33%), Pseudomonas aeruginosa (24%), Acinetobacter spp. (22%) e Klebsiella pneumoniae (5%). Conclusão: A avaliação microbiológica do primeiro ano de funcionamento da UTQ da EPM/ UNIFESP revelou que, embora o agente mais prevalente tenha sido a CoNS, bacilos Gram negativos ainda são muito prevalentes, como a Pseudomonas aeruginosa e a Acinetobacter baumannii. Apesar de pouco tempo de operação, observou-se um grande número de microrganismos multirresistentes, que pode ser explicado por longa exposição a agentes antimicrobianos e alta taxa de transferência de outros hospitais.

PURPOSE: To evaluate the expression profile of genes related to Toll Like Receptors (TLR) pathways of human Primary Epidermal keratinocytes of patients with severe burns. METHODS: After obtaining viable fragments of skin with and without burning, culture hKEP was initiated by the enzymatic method using Dispase (Sigma-Aldrich). These cells were treated with Trizol(r) (Life Technologies) for extraction of total RNA. This was quantified and analyzed for purity for obtaining cDNA for the analysis of gene expression using specific TLR pathways PCR Arrays plates (SA Biosciences). RESULTS: After the analysis of gene expression we found that 21% of these genes were differentially expressed, of which 100% were repressed or hyporegulated. Among these, the following genes (fold decrease): HSPA1A (-58), HRAS (-36), MAP2K3 (-23), TOLLIP (-23), RELA (-18), FOS (-16), and TLR1 (-6.0). CONCLUSIONS: This study contributes to the understanding of the molecular mechanisms related to TLR pathways and underlying wound infection caused by the burn. Furthermore, it may provide new strategies to restore normal expression of these genes and thereby change the healing process and improve clinical outcome.

PURPOSE: To evaluate KGF and human beta defensin-4 (HBD-4) levels produced by dermic fibroblasts and keratinocytes cultivated from burned patients' skin samples. METHODS: Keratinocytes and fibroblasts of 10 patients (four major burns, four minor burns and two controls) were primarily cultivated according to standard methods. HBD-4 and KGF genes were analyzed by quantitative PCR. RESULTS: In fibroblasts, KGF gene expression was 220±80 and 33.33±6.67 (M±SD; N=4), respectively for major and minor burn groups. In keratinocytes, KGF gene expression was 11.2±1.9 and 3.45±0.37 (M±SD; N=4), respectively for major and minor burn groups. In fibroblasts, HBD-4 gene expression was 15.0±4.0 and 11.5±0.5 (M±SD; N=4), respectively for major and minor burn. In keratinocyte, HBD-4 gene expression was 0.0±0.0 and 13.4±4.8 (M±SD; N=4), respectively for major and minor burn. CONCLUSIONS: KGF expression was increased in burn patient fibroblasts compared to control group. In keratinocytes culture, KGF suppression is inversely proportional to burn extension; it is active and increased in major burn but decreased in minor burn. HBD-4 expression was increased in fibroblasts and decreased in keratinocytes from all burned patients.

PURPOSE: To evaluate the level of cytokines and keratinocyte growth factor (KGF) or Fibroblast Growth Factor 7 (FGF-7) in the culture medium of cultured human dermal fibroblasts from patients with large burn in comparison to small burn. METHODS: Fibroblasts of 10 patients (four large burns, four small burns and two controls) were initiated by the enzymatic method using collagenase. Cytokines and KGF in the supernatant of the culture medium was measured by, respectively, flow cytometry using Cytometric Bead Array Human Inflammation kit (CBA, BD Biosciences, USA) and the enzyme immunoassay method using the Quantikine (r) Human KGF. The experiments were performed in triplicate. RESULTS: The expression of IL-12 protein in patients with large burns showed a tendency to increase. IL- 6, IL- 10, and IL- 1beta were observed no difference. For IL - 8, TNF - alpha and KGF was observed a significant difference between the expression in large and small burned patient. CONCLUSION: That IL-8, TNF-alpha and KGF showed higher expression in cultured fibroblasts of large burned patients.

PURPOSE: This study is a systematic literature review and meta-analysis concerning the use of a testosterone synthetic analog, oxandrolone, and its use in severe adult burns. METHODS: Randomized prospective clinical studies, in English, Portuguese or Spanish, were sought on the following databases: MEDLINE, COCHRANE, EMBASE and LILACS. There was no restriction in relation to the publication date. RESULTS: This search produced 24 studies on MEDLINE and twelve articles were presented on the COCHRANE database .Sixteen were excluded due to the title not being related to this search or by including children. Of the eigth residual studies, after adaptation to the inclusion criteria, only four were selected. After analyzing the results, two were discarded since they did not present adequate patient characterization and the facts on these articles were analyzed differently from the others, hindering the meta-analysis. CONCLUSION: The analysis of the available data demonstrated significant benefits (p<0.05) considering lesser loss of corporal mass, lesser nitrogen loss, and shorter donor area healing time, when Oxandrolone was used, comparatively with the control group (placebo or not).

PURPOSE: To evaluate the role of transforming growth factor beta 1 (TGF-β1) on the induced osteogenic differentiation of human dermal fibroblasts. METHODS: We performed four groups with cultured dermal fibroblasts according to the culture medium: CONTROL (DMEM culture medium); TGF-β1 (DMEM culture medium with 10 ng/ml of TGF-β1); OSTEOG (DMEM culture medium with 0.5 µg/ml of ascorbic acid, 10 mmol/l of β-glycerophosphate and 10 nmol/L of dexamethasone); and OSTEOG/TGF-β1 (osteogenic medium with 10 ng/ml of TGF-β1). Alkaline phosphatase (ALP) activity and the amount of osteocalcin (OC) in the supernatant, as well as the capability to form calcium phosphate deposits, were analysed for 28 days RESULTS: There were significant differences (p<0.05) between CONTROL and TGF-β1 groups in comparison with OSTEOG and OSTEOG/TGF-β1 groups in the ALP activity and OC amount. Although, both osteogenic groups had the same behavior with regard the expression curve during the experimental time, the OSTEOG/TGF-β1 group achieved significantly higher ALP and OC levels and showed no significant difference in the levels of mineralized deposits and in comparison with the levels found in the OSTEOG group. CONCLUSION: The addition of transforming growth factor beta 1 to the osteogenic culture medium increased the activity of alkaline phosphatase and the amount of osteocalcin, but TGF-β1 did not alter the presence of mineralized calcium phosphate deposits.

PURPOSE: Evaluate the expression profile of genes related to Innate and Adaptive Immune System (IAIS) of human Primary Epidermal keratinocytes (hPEKP) of patients with severe burns. METHODS: After obtaining viable fragments of skin with and without burning, culture hKEP was initiated by the enzymatic method using Dispase (Sigma-Aldrich). These cells were treated with Trizol(r) (Life Technologies) for extraction of total RNA. This was quantified and analyzed for purity for obtaining cDNA for the analysis of gene expression using specific IAIS PCR Arrays plates (SA Biosciences). RESULTS: After the analysis of gene expression we found that 63% of these genes were differentially expressed, of which 77% were repressed and 23% were hyper-regulated. Among these, the following genes (fold increase or decrease): IL8 (41), IL6 (32), TNF (-92), HLA-E (-86), LYS (-74), CCR6 (- 73), CD86 (-41) and HLA-A (-35). CONCLUSIONS: This study contributes to the understanding of the molecular mechanisms underlying wound infection caused by the burn. Furthermore, it may provide new strategies to restore normal expression of these genes and thereby change the healing process and improve clinical outcome.

PURPOSE: To evaluate the gene expression of KGF, TNF-alpha and IL-1 beta in skin fibroblasts and keratinocytes cultured from burned patients. METHODS: Three patients with large burns and three patients with small burns, as well as two controls, were included. The cell culture was initiated by the enzymatic method. After extraction and purification of mRNA, qPCR was used to assess the gene expression of KGF, TNF-alpha and IL-1 beta. RESULTS: The expression of KGF was increased on average 220-fold in large burns and 33.33-fold in small burns in fibroblasts, and 11.2-fold in large burns and 3.45-fold in small burns in keratinocytes compared to healthy patients (p<0.05). Expression of TNF-alpha was not observed. IL-1 beta is down-regulated in fibroblasts of burned patients, and much more repressed in small burns (687-fold, p<0.05). In keratinocytes, the repression of IL-1 beta expression occurs in patients with small burns (28-fold), while patients with large burns express this gene intensively (15-fold). CONCLUSIONS: The study showed a quantitative pattern in the expression of KGF gene, which is more expressed according to the size of the burn. TNF-alpha was not expressed. A qualitative pattern in the expression of IL-1 beta gene was demonstrated.

PURPOSE: To assess the effects of unripe Musa sapientum peel on the healing of surgical wounds in rats. METHODS: One hundred and twenty Wistar rats were divided into two treatment groups of 60 animals each: the control group (gel without the active ingredient) and experimental group (4% Musa sapientum peel gel). A 4 x 4 cm surgical wound was created on the back of each animal. The wound was cleaned daily with 0.9% saline, treated with 4% gel or natrosol gel (control), and covered with gauze. Animals from both groups were sacrificed after seven, 14 and 21 days of treatment; the tissue from the wound site was removed together with a margin of normal skin for histological analysis. RESULTS: No significant differences in wound contraction rates (p=0.982) were found between time points (seven, 14 and 21 days of treatment) in both groups. However, a significantly higher wound contraction rate was observed in the control group on day 21 compared with the experimental group (p=0.029). There were no significant differences in histomorphological features between groups. The experimental group showed an increased number of polymorphonuclear cells on day 7, with a significant reduction on day 21 (p=0.026). CONCLUSION: The use of 4% unripe Musa sapientum peel gel on surgical wounds in rats resulted in an increased number of polymorphonuclear cells on day 7, reduced wound contraction, reduced vascular proliferation and increased concentration of collagen fibers on day 21.