The role of vitamin C in the gene expression of oxidative stress markers in fibroblasts from burn patients

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.

Aldefluor protocol to sort keratinocytes stem cells from skin

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.

Keratinocyte growth factor and the expression of wound-healing-related genes in primary human keratinocytes from burn patients

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.

Quality of Life and Sleep of a Multidisciplinary Team in the Burns Treatment Unit of Hospital São Paulo / Qualidade de vida e de sono da equipe multidisciplinar na Unidade de Tratamento de Queimaduras do Hospital São Paulo

Introdução: O objetivo foi avaliar a qualidade de vida e do sono da equipe multidisciplinar da Unidade de Tratamento de Queimaduras. Métodos: O questionário Short Form-36 Medical Outcomes Survey (SF-36) e o Pittsburgh Sleep Quality Index (PSQI) foram utilizados em 50 trabalhadores da unidade. Resultados: A limitação dos aspectos físicos, da saúde geral, da vitalidade e da limitação da saúde social e mental foram significativamente correlacionada com o sono (p<0,05). Conclusões: Esse ambiente de trabalho não é propício para o autocuidado dos trabalhadores; ao contrário, é um lugar de mobilização para emoçõoes e para o estresse.

Background: The aim of this study was to assess the quality of life and sleep of the multidisciplinary team of the Burn Unit. Methods: The Short Form-36 Medical Outcomes Survey (SF-36) and Pittsburgh Sleep Quality Index (PSQI) were used in 50 workers. Results: The limitation of the physical aspects, the general health, vitality, and limitation of social and mental health were significantly correlated with sleep (p<0.05). Conclusions: The work environment is not conducive to workers' self-care; rather, it is a mobilizing place for emotions and stress.

Perfil microbiológico da unidade de queimaduras da EPM/UNIFESP, São Paulo, Brasil / Microbiological surveillance of burn unit of EPM/UNIFESP in São Paulo, Brazil

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.

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.

Toll like receptors gene expression of human keratinocytes cultured of severe burn injury

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. .

Human beta defensin-4 and keratinocyte growth factor gene expression in cultured keratinocyte and fibroblasts of burned patients

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. .

Innate and adaptive immunity gene expression of human keratinocytes cultured of severe burn injury

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. .

Oxandrolone use in adult burn patients. Systematic review and meta-analysis

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). .

Keratinocyte growth factor, interleukins (1 beta, 6, 8, 10, 12), and tumor necrosis factor alpha in culture medium of dermal fibroblast of 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. .

TGF-β1 on induced osteogenic differentiation of human dermal fibroblast

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. .

Keratinocyte growth factor, tumor necrosis factor-alpha and interleukin-1 beta gene expression in cultured fibroblasts and keratinocytes from burned patients

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.

Unripe Musa sapientum peel in the healing of surgical wounds in rats

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.

AZT on telomerase activity and cell proliferation in HS 839.T melanoma cells / AZT na atividade da telomerase e na proliferação de células de melanoma HS 839.T

PURPOSE: To evaluate telomerase activity and proliferation of HS839.T melanoma cells, subjected to the action of AZT. METHODS: Cells were grown in triplicate, AZT at different concentrations: 50, 100 and 200μM, was added and left for 24 and 48 hours, and its effects were compared with the control group. Telomerase activity was detected by PCR and cell proliferation was evaluated by MTT. RESULTS: After 24 hours, there was no inhibition of cell proliferation or telomerase activity when compared to the control group. After 48 hours, there was a momentary decrease, suggesting that the cell lines used in this study are sensitive to AZT, but quickly recover both the enzyme activity and cell proliferation. CONCLUSION: The action of AZT on the melanoma cells studied, at the concentrations and times tested, did not inhibit telomerase activity nor affect cell proliferation.

OBJETIVO: Avaliar a atividade da telomerase e da proliferação de células de melanoma HS839.T submetidas à ação do AZT. MÉTODOS: As células foram cultivadas, em triplicata, com diferentes concentrações de AZT: 50, 100 e 200µM, por 24h e 48h, seus efeitos comparados com o grupo controle. A atividade da telomerase foi detectada por PCR e a proliferação celular avaliada por MTT. RESULTADOS: No tempo de 24 horas, não houve inibição da proliferação celular e da atividade da telomerase em comparação com o grupo controle. No período de 48 horas, houve uma diminuição momentânea, sugerindo que as células das linhagens utilizadas neste estudo são sensíveis ao AZT, mas que recuperam a atividade enzimática e proliferativa. CONCLUSÃO: Nas células de melanoma HS839.T estudadas e nas concentrações e tempos propostos, a ação do AZT não inibiu a atividade da telomerase e não afetou a proliferação celular.

Portland cement with additives in the repair of furcation perforations in dogs / Cimento Portland com aditivos na reparação de perfurações radiculares em cães

PURPOSE: To evaluate the use of Portland cements with additives as furcation perforation repair materials and assess their biocompatibility. METHODS: The four maxillary and mandibular premolars of ten male mongrel dogs (1-1.5 years old, weighing 10-15 kg) received endodontic treatment (n=80 teeth). The furcations were perforated with a round diamond bur (1016 HL). The perforations involved the dentin, cementum, periodontal ligament, and alveolar bone. A calcium sulfate barrier was placed into the perforated bone to prevent extrusion of obturation material into the periradicular space. The obturation materials MTA (control), white, Type II, and Type V Portland cements were randomly allocated to the teeth. Treated teeth were restored with composite resin. After 120 days, the animals were sacrificed and samples containing the teeth were collected and prepared for histological analysis. RESULTS: There were no significant differences in the amount of newly formed bone between teeth treated with the different obturation materials (p=0.879). CONCLUSION: Biomineralization occurred for all obturation materials tested, suggesting that these materials have similar biocompatibility.

OBJETIVO: Avaliar o uso de cimentos Portland aditivados na reparação de perfurações radiculares e a biocompatibilidade destes materiais. MÉTODOS: Oitenta pré-molares, quatro da arcada dentária superior e quatro da arcada inferior de 10 cães machos, sem raça definida, com idade em torno de um a um ano e meio, pesando entre 10 e 15 kg foram submetidos a tratamento endodôntico, sendo realizadas perfurações nas furcas com broca de diamante 1016 HL. A cavidade envolveu dentina e cemento, como também periodonto e o osso alveolar. Na porção óssea da obturação, barreira de sulfato de cálcio foi utilizada evitando extravasamento do cimento para o espaço periodontal. Foi realizada a distribuição randomizada dos cimentos MTA (controle), Portland tipo II, Portland tipo V e Portland branco estrutural nas obturações. Os dentes foram restaurados com resina composta. Após 120 dias realizou-se eutanásia, retirada dos dentes, preparação e análise histológica. RESULTADOS: Entre os cimentos não houve diferença estatística significante quanto à neoformação óssea (p=0,879). CONCLUSÃO: Ocorreu biomineralização com os diferentes cimentos usados no estudo, sugerindo que estes são similares em termos de biocompatibilidade.

Rat an experimental model for burns: A systematic review / Rato como modelo experimental de queimadura: Revisão sistemática

PURPOSE: To revise and systematize scientific knowledge of the experimental model for cutaneous burns in rats. METHODS: A bibliographical review from 2008 up to January 2011 in PubMed, EMBASE and LILACS was undertaken. Were used the keywords: animal models, burns and rats. 221 studies were identified, and 116 were selected. RESULTS: It was found that: 54/86 (62.7%) had third degree burns; 55/73 (75.3%) studied the back; 45/78 (57.6%) used heated water and 27/78 (35.9%) incandescent instruments; 39/78 (50%) studied systemic effects; 22/71 (31%) used ketamine associated with xylazine; 61/64 (95.3%) performed depilation with appropriate equipment; 36/72 (50%) used microscopy; more than 50% did not describe analgesia or antibiotics during the postoperative period; in 42/116 (36.2%) postoperative fluid therapy was performed; and the time interval after the burn, up to the beginning of the results analysis varied from 7s up to four weeks. Legislation issues on burn experiments are discussed. CONCLUSION: The hot water was the main method to induce burns those of third degree on the back, with anesthesia using ketamine and xylazine, after depilation. These were evaluated microscopically, without using analgesia or an antibiotic during the postoperative period. The studies were not very reproducible.

OBJETIVO: Revisar e sistematizar o conhecimento científico do modelo experimental em queimadura da pele em ratos. MÉTODOS: Revisão da literatura foi realizada de 2008 a Janeiro de 2011 na PubMed, EMBASE e LILACS. Os descritores usados foram: modelo animal, queimadura e ratos. 221 estudos foram identificados e 116 foram selecionados. RESULTADOS: Foi encontrado que: 54/86 (62,7%) tinham queimadura de terceiro grau; 55/73 (75,3%) estudaram o dorso; 45/78 (57.6%) usaram líquido aquecido e 27/78 (35,9%) usaram instrumento incandescente; 39/78 (50%) estudaram efeitos sistêmicos; 22/71 (31%) usaram ketamina associada a xilazina; 61/64 (95,3%) realizaram depilação com equipamento apropriado; 36/72 (50%) usaram microscopia; mais de 50% não descreveram uso de analgésicos ou antibióticos durante o período pós-operatório; em 42/116 (36,2%) foi realizada reposição hídrica pós-operatória; e o intervalo de tempo após a queimadura e a análise variou de 7s a quatro semanas. Aspectos legais sobre experimentos em queimaduras foram discutidos. CONCLUSÃO: Líquido aquecido foi o principal método para induzir queimadura de terceiro grau no dorso do animal, com anestesia usando quetamina e xilazina, após depilação, avaliados por microscopia, sem uso de analgesia ou antibióticos. Os estudos não são reprodutíveis.

Ácido ascórbico intracelular diminui a indução à senescência celular, mas não a apoptose em fibroblastos expostos a concentrações subtóxicas de H2O2 / Intracellular ascorbic acid reduces the induction of cellular senescence, but not apoptosis in fibroblastos exposed to sub toxic concentrations of H2O2

Introdução: As espécies reativas de oxigênio (EROs) são produzidas durante o metabolismo normal das células, tendo funções fisiológicas, como a sinalização celular. Porém, em uma situação que leve o organismo à produção exagerada de EROs, temos o chamado estresse oxidativo, que tem ação deletéria às células, podendo levar a apoptose ou senescência celular. A vitamina C é um dos principais a gentes antioxidantes do organismo, atuando em todas as formas de estresse oxidativo. Método: Isolamento e cultivo de fibroblastos humanos dérmico em seis grupos: controle, Vitamina C +, Vitamina C -, H2O2, Vitamina C + H2O2, Vitamina C - H2O2. Os fibroblastos foram submetidos ao estresse oxidativo pela suplementação de H2O2 ao meio de cultura por 2 horas. Foram avaliadas proliferação pelo MTT, senescência celular pela marcação da enzima beta-galoctosidase, apoptose celular e liberação de radicais livres por citometria de fluxo. Resultados: O peróxido de hidrogênio aumentou significantemente a senescência celular e a apoptose nos fibroblastos, enquanto a vitamina C diminuiu significantemente a indução a senescência celular somente no estado intracelular. Conclusões: O ácido ascórbico não protegeu os fibroblastos humanos dérmicos cultivados contra o estresse oxidativo induzido pelo H2O2. O ácido ascórbico intracelular levou à diminuição da indução à senescência celular.

Introduction: Reactive oxygen species (ROS) are produced during normal metabolism of cells, with physiological functions such as cell signaling. But in a situation that causes the body to ROS overproduction, we have the so-called oxidative stress, which has deleterious effects on cells, leading to apoptosis or cellular senescence. Vitamin C is one of the main antioxidants in the body, working in all forms of oxidative stress. Methods: Isolation and culture of human dermal fibroblasts in six groups: control, + Vitamin C, - Vitamin C, H2O2, H2O2 + Vitamin C, H2O2 – Vitamin C. The fibroblasts were subjected to oxidative stress by supplements H2O2 to the culture medium for 2 hours. Were evaluated by MTT proliferation assay, cellular senescence by marking the enzyme beta-galactosidade, apoptosis and release of free radicals by flow cytometry. Results: Hydrogen peroxide significantly increased cellular senescence and apoptosis in fibroblasts, while vitamin C decreased significantly the induction of cellular senescence, just in the intracellular state. Conclusions: Ascorbic Acid did not protect cultured human dermal fibroblasts against oxidative stress induced by H2O2. Ascorbic acid has led to decreased intracellular induction of cellular senescence.

Gel from unripe Musa sapientum peel to repair surgical wounds in rats / Gel da casca de Musa sapientum verde no reparo de lesões operatórias em ratos

PURPOSE: To determine the optimum concentration of a gel obtained from unripe banana (Musa sapientum) peel for wound treatment in rats. METHODS: A randomized triple blind study was conducted with 40 Wistar rats, which were divided into 4 groups: CG, control group; G2 percent, 2 percent gel concentration group; G4 percent, 4 percent gel concentration group; and G10 percent, 10 percent gel concentration group. The banana peel gel was applied daily, for 7 days, to a 4-cm² wound created on the back of each animal of all groups. After this period, the wounds were biopsied. Statistical analysis was carried out using the Kruskal-Wallis test complemented by the Student-Newman-Keuls test. RESULTS: Macroscopic examination revealed that partial epithelialization occurred in all groups. Wound contraction was also observed in all groups and ranged from 1.38 to 1.57 mm in the study groups, and from 1.03 to 1.10 mm in the control group, with significant differences (p < 0.05) between the groups: CG and G10 percent, G2 percent and G4 percent, G2 percent and G10 percent. The interquartile deviation was smaller between the groups CG and G4 percent. CONCLUSION: The 4 percent gel obtained from unripe banana peel (G4 percent) resulted in better epithelialization of wounds healed by secondary intention compared with other gel concentrations.

OBJETIVO: Avaliar a concentração ideal do gel da casca de Musa sapientum verde no tratamento de feridas em ratos. MÉTODOS: Estudo randomizado, triplo cego, com 40 ratos da linhagem Wistar divididos em quatro grupos: GC controle, G2 por cento gel a 2 por cento, G4 por cento gel a 4 por cento, G10 por cento gel a 10 por cento. Realizou-se aplicação diária do gel nas diferentes concentrações, durante sete dias, em uma ferida de 4 cm² realizada no dorso de cada rato. Após este período, as lesões foram biopsiadas. Para analise dos dados utilizou-se o teste de Kruskal-Wallis complementado pelo teste de Student-Newman-Keuls. RESULTADOS: Os achados macroscópicos demonstraram reepitelização parcial em todos os grupos. A contração da área da ferida variou entre 1,38 a 1,57 mm nos grupos de estudo, e entre 1,03 a 1,10 mm no grupo controle. Houve diferença significante (p < 0,05) entre os grupos: GC e G10 por cento, G2 por cento e G4 por cento, G2 por cento e G10 por cento, sendo o desvio interquartílico menor entre os grupos GC e G4 por cento. CONCLUSÃO: O gel a 4 por cento da casca de M. sapientum verde promoveu maior área de epitelização em feridas com cicatrização por segunda intenção, em relação ao gel nas outras concentrações testadas.

Root perforations treatment using mineral trioxide aggregate and Portland cements / Cimentos de agregado trióxido mineral e Portland no tratamento de perfurações radiculares

PURPOSE: Clinical, radiological and histological evaluation of root perforations treated with mineral trioxide aggregate (MTA) or Portland cements, and calcium sulfate barrier. METHODS: One molar and 11 premolar teeth of a male mongrel dog received endodontic treatment and furcations were perforated with a high-speed round bur and treated with a calcium sulfate barrier. MTA, Portland cement type II (PCII) and type V (PCV), and white Portland cement (WPC) were used as obturation materials. The teeth were restored with composite resin and periapical radiographs were taken. The animal was euthanized 120 days post-surgery for treatment evaluation. RESULTS: Right lower first premolar (MTA), right lower third premolar (PCV), left lower second premolar (MTA), and right lower second premolar (WPC): clinically normal, slightly radio-transparent area on the furcation, little inflammatory infiltrate, and new-bone formation. Left lower third premolar (PCII), right upper first premolar (WPC), right upper third premolar (PCII), and left upper first molar (PCV): clinically normal, radiopaque area on the furcation, and new-bone formation. Right upper second premolar (MTA), left upper second premolar (WPC), left upper third premolar (PCII): presence of furcation lesion, large radiolucent area, and intense inflammatory infiltrate. CONCLUSION: All obturation materials used in this study induced new-bone formation.

OBJETIVO: Avaliar clínica, radiológica e histologicamente perfurações radiculares tratadas com MTA e cimentos Portland, com barreira de sulfato de cálcio. MÉTODOS: A amostra foi constituída por 11 dentes pré-molares e um molar de cão macho, sem raça definida. Após tratamento endodôntico realizaram-se perfurações nas furcas com broca esférica de alta rotação e barreira de sulfato de cálcio. Foram utiliados os cimentos MTA, Portland tipo II (CPII), Portland tipo V (CPV) e Portland branco estrutural (CPB). Os dentes foram restaurados com resina composta e submetidos a radiografias periapicais. O animal foi confinado por 120 dias e submetido à eutanásia para avaliações. RESULTADOS: Primeiro pré-molar inferior direito (MTA), terceiro pré-molar inferior direito (CPV), segundo pré-molar inferior esquerdo (MTA) e segundo pré-molar inferior direito (CPB): clinicamente normal; leve área radiotransparente na furca; pequeno infiltrado inflamatório e neoformação óssea. Terceiro pré-molar inferior esquerdo (CPII), primeiro pré-molar superior direito (CPB), terceiro pré-molar superior direito (CPII) e primeiro molar superior esquerdo (CPV): clinicamente normal; área radiopaca na furca; neoformação óssea. Segundo pré-molar superior direito (MTA), segundo pré-molar superior esquerdo (CPB) e terceiro pré-molar superior esquerdo (CPII): clinicamente com lesão na furca; intensa área radiolucida; infiltrado inflamatório intenso. CONCLUSÃO: Todos os cimentos induziram a neoformação óssea.