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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.

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.

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

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.

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.

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.

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%, 2% gel concentration group; G4%, 4% gel concentration group; and G10%, 10 % 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%, G2% and G4%, G2% and G10%. The interquartile deviation was smaller between the groups CG and G4%. CONCLUSION: The 4% gel obtained from unripe banana peel (G4%) 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% gel a 2%, G4% gel a 4%, G10% gel a 10%. 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%, G2% e G4%, G2% e G10%, sendo o desvio interquartílico menor entre os grupos GC e G4%. CONCLUSÃO: O gel a 4% 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.

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.

PURPOSE: To evaluate effects of oxidative stress and supplementation of keratinocyte growth factor (KGF) on cultivated human keratinocytes. METHODS: Oxidative stress was produced through addition of hydrogen peroxide (H2O2) to the culture medium. Cultivated human keratinocytes were divided in 4 groups: Group control (G C), Group KGF (G KGF), Group H2O2 (G H2O2), Group H2O2 and KGF (G H2O2-KGF). Each experiment was accomplished with the same lineage cultivated keratinocytes, in triplicate. Cell viability was evaluated by trypan blue exclusion assay. RESULTS: The results showed that the culture medium supplemented with KGF presented a small rate of cell viability when compared to cells only in culture medium (pOBJETIVO: Avaliar os efeitos do estresse oxidativo e da suplementação do fator de crescimento de queratinócitos (KGF) em queratinócitos humanos cultivados. MÉTODOS: O estresse oxidativo foi produzido através da adição de peróxido de hidrogênio (H2O2) ao meio de cultura. Os queratinócitos humanos cultivados foram divididos em quatro grupos: grupo controle (G C), grupo KGF (G KGF), grupo H2O2 (G H2O2), grupo H2O2 e KGF (G H2O2-KGF). Cada experimento foi realizado com a mesma linhagem celular, em triplicata. A viabilidade celular foi avaliada pelo ensaio da exclusão do azul de tripan. RESULTADOS: Os resultados mostraram que o meio de cultura suplementado com o KGF apresentou menor taxa de viabilidade celular quando comparado às células do grupo controle (p

The use of cell culture methods in Plastic Surgery opened a new horizon in the research of cellular mechanisms of proliferation and biosynthesis functions. Several types of cells have been investigated in the cutaneous compartment. Keratinocytes and fibroblasts have been studied aiming the possibility of developing biomaterial for skin substitution. The present study describes the standardization for the development of fibroblast primary culture, its utilization in experiments and its storage.A utilização de métodos de cultura de células em Cirurgia Plástica abriu um novo horizonte na pesquisa dos mecanismos celulares de proliferação e biossíntese. Vários tipos de células têm sido investigados no compartimento cutâneo. Os queratinócitos e os fibroblastos têm sido estudados objetivando-se o desenvolvimento de biomateriais para substituição de pele. O presente estudo descreve uma padronização metodológica para o estabelecimento de uma cultura primária de fibroblastos, sua utilização em experimentos e seu armazenamento.

In Plastic Surgery, cell culture represents the perspective of studying cellular mechanisms that guide the healing process of several tissues. Some steps of the healing process depend on physical factors as the tissular partial pressure of O2. In cell culture, it is possible to submit cells to hypoxic enviroment. The present study reports an alternative method at low cost for the establishment of hypoxic environment in cell culture flasks.A cultura de células, na Cirurgia Plástica, representa uma perspective para o estudo dos mecanismos celulares que norteiam o processo cicatricial de diversos tecidos. Algumas etapas do processo de cicatrização dependem de fatores físicos como a pressão parcial de O2. Em uma cultura de células, é possível submeter células a um ambiente hipóxico. O presente estudo relata um método alternativo de baixo custo para o estabelecimento de um ambiente hipóxico em frascos de cultura de células.

One of the most used animal models of cultured keratinocytes autografting is based on xenografting of human keratinocytes to the rat or athymic mice, immunological neutral recipient that acts as biological carrier. It could be studied in this model many facts that occur after transplant without the ethical aspect in the clinical study. The proposition of the experimental model is related to the sequence of the total or partial skin transplant, as autografting or xenografting, cultured or not, to the back of athymic mice. The model presents the possibility of study in vivo athymic animal, when the in vivo study in anima nobili is not ethical. It permits the xenografting evaluation of cultured cells graft or of the genetically modified cells and of the association of the cultured cells and the dermal substitutes, the composite grafts, and of the autografting.Um dos modelos animais mais utilizados de auto-enxertia de queratinócitos cultivados é baseado em xeno-enxerto de queratinócitos humanos em rato atímico, um receptor imunologicamente neutro que atua como carreador biológico. Muitos fatos podem ser estudados nesse modelo que acontecem após o transplante sem os aspectos éticos do estudo clínico. A proposição do modelo experimental esta relacionada a sequência do transplante de pele parcial ou total como auto-enxerto ou xeno-enxerto, cultivado ou não, no dorso do rato atímico. O modelo apresenta a possibilidade do estudo in vivo do animal atímico, quando o estudo in vivo em anima nobili não é considerado ético. Isso permite a avaliação do xeno-enxerto de células humanas normais ou modificadas geneticamente modificadas cultivadas e a associação de células cultivadas e substitutes dérmicos, de enxertos compostos e de auto-enxerto.

One of the most used animal models of cultured keratinocytes autografting is based on xenografting of human keratinocytes to the rat or athymic mice, immunological neutral recipient that acts as biological carrier. It could be studied in this model many facts that occur after transplant without the ethical aspect in the clinical study. The proposition of the experimental model is related to the sequence of the total or partial skin transplant, as autografting or xenografting, cultured or not, to the back of athymic mice. The model presents the possibility of study in vivo athymic animal, when the in vivo study in anima nobili is not ethical. It permits the xenografting evaluation of cultured cells graft or of the genetically modified cells and of the association of the cultured cells and the dermal substitutes, the composite grafts, and of the autografting.

In the study of Plastic Surgery, cell culture may be used at experimental level in researches concerning biosynthesis functions of skin cells such as fibroblasts, keratinocytes, adipocytes, chondrocytes and osteocytes. The present study reports an experimental model for estimation of collagen in cell cultures using chromogenic precipitation reaction with an especific dye (Sirius Red).Dentro do estudo da Cirurgia Plástica, a cultura de células pode ser utilizada em experimentos relativos às funções de biossíntese de células relacionadas com a pele tais como fibroblastos, queratinócitos, adipócitos, condrócitos e osteócitos. O presente estudo relata modelo experimental para a mensuração estimada do colágeno em cultura de células utilizando-se uma reação de precipitação cromogênica com um corante específico (Sirius Red).

The bioengineering research is essential in the development of ideal combination of biomaterials and cultured cells to produce the permanent wound coverage. The experimental model of cultured keratinocytes presents all steps of the culture, since the isolation of the keratinocytes, preparation of the human acellular dermis, preparation of the composite skin graft and their elevation to the air-liquid interface. The research in cultured keratinocytes model advances in two main ways: 1. optimization of the methods in vitro to the skin cells culture and proliferation and 2. developing biomaterials that present similar skin properties.A pesquisa em bioengenharia é primordial no desenvolvimento da combinação ideal de biomateriais e células cultivadas para produzir a cobertura definitiva das lesões. O modelo experimental da cultura de queratinócitos apresenta toda as etapas do cultivo, desde o isolamento dos queratinócitos, preparação da derme acelular humana, do enxerto composto e da sua elevação à interface ar-líquido. A pesquisa em modelo de cultura de queratinócitos desenvolve-se em duas vias principais: 1. otimização dos métodos in vitro para cultivo e proliferação de células da pele e 2. desenvolvimento de biomateriais que mimetizem as propriedades da pele.

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 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

PURPOSE: To evaluate effects of oxidative stress and supplementation of keratinocyte growth factor (KGF) on cultivated human keratinocytes. METHODS: Oxidative stress was produced through addition of hydrogen peroxide (H2O2) to the culture medium. Cultivated human keratinocytes were divided in 4 groups: Group control (G C), Group KGF (G KGF), Group H2O2 (G H2O2), Group H2O2 and KGF (G H2O2-KGF). Each experiment was accomplished with the same lineage cultivated keratinocytes, in triplicate. Cell viability was evaluated by trypan blue exclusion assay. RESULTS: The results showed that the culture medium supplemented with KGF presented a small rate of cell viability when compared to cells only in culture medium (p

Establish an experimental model of metastatic melanoma primary culture cells from fine-needle aspiration biopsies (FNAB).Two metastatic melanoma primary culture cells from (FNAB) have been developed from patients who had been submitted to excision of metastatic lesions and they were identified by immunohistochemical analyses using S-100 and MB-45 melanoma markers.The culture diagnostic was confirmed by immunohistochemical. The technique of FNAB offers the advantage of providing a sequential analysis of the same tumor nodules throughout treatment, and is a minimally invasive procedure with almost no associated morbidity.The establishment of metastatic melanoma primary culture from FNAB samples showed a viable technique.