La régénération de la nageoire caudale et de la peau chez le poisson zèbre impliquent la formation d’un tissu de remplacement, structurellement et fonctionnellement identique à celui précédant la lésion. Ces mécanismes nécessitent la mise en place et la prolifération de cellules progénitrices, capables de reformer lors d’une phase de « patterning » les tissus lésés.Au cours de ma thèse j’ai étudié les évènements prolifératifs qui permettent cette néo-formation des tissus. Dans le cas de la nageoire, mes travaux ont conduit à la mise en évidence d’un mécanisme commun entre régénération et développement de croissance saltatoire des segments osseux. Ils expliquent en partie le retour à la forme bilobée observée lors de la repousse. De la même façon, dans le cas de la peau, l’avènement de la prolifération dans le derme et dans l’épiderme précède la mise en place des signaux, communs au développement, requis pour la distribution et la formation des écailles.De plus, j’ai effectué une caractérisation précise de l’ensemble du processus cicatriciel chez le poisson zèbre, conduisant à la formation d’une peau intégralement régénérée. Au cours de la cicatrisation, la fermeture de la blessure est complétée rapidement en quelques heures par migration épidermique. Une fois la zone lésée fermée, un mécanisme de morphogénèse de la peau est réactivé chez l’adulte. La prolifération cellulaire est présente simultanément dans le derme et l’épiderme. Elle n’est déclenchée qu’après la mise en place de l’assise basale de l’épiderme. Dans celui-ci, elle affecte d’une part des cellules éparses au sein de la couche basale et d’autre part la majorité des cellules de la couche intermédiaire. Ces derniers travaux suggèrent que chez les téléostéens, les cellules souches épidermiques sont localisées dans la couche basale, alors que la prolifération des cellules transientes a lieu dans la couche intermédiaire. D’autre part, ils mettent en évidence un processus commun de cicatrisation rapide en milieu liquide impliquant une fermeture de la blessure par néo-épithélialisation, semblable au cas de la cornée et de la muqueuse orale chez les mammifères. / Caudal fin and skin regeneration in zebrafish both require new tissues formation, structurally and functionally identical to the former ones. They imply the formation and especially the proliferation of progenitor cells, and then during a patterning phase, they differentiate into a well-organized structure.During my PhD, I have studied in zebrafish model the proliferative events that conduct to the neoformation of caudal fin in one hand and the proliferative events implicated in the cutaneous wound healing in the other hand.The first part of this work supports the evidences of a common saltatory growth mechanism in both regeneration and development of caudal fin bony rays, and that the bi-lobed shape restoration of the fin could be a consequence of this bony segment salutary growth.During skin wound healing, proliferation is necessary in order to allow dermis and epidermis neo-morphogenesis and these events are over before the scale formation is initiated. In the second part of this study I characterized the entire skin wound healing process in the zebrafish model, from the wound closure to a fully regenerated skin including appendages. According to my results, the wound closure is a very fast event completed only in a few hours and it occurs only by epidermal cells migration. Cellular proliferation was detected after complete wound closure and once the epidermal basal layer differentiation is achieved. Cell proliferation appears to be restricted to a few basal cells whereas the major proliferation is detected in the intermediates layers of the epidermis.Taken together, these results suggest that in teleosteans, the epidermal stem cells and transient cells might be respectively located in the basal and intermediate layers. Moreover, there might be a common process due to aquatic environment, leading to a fast wound closure by re-epithelialization, between teleost skin and mammal’s cornea as well as oral mucosa.

Made available in DSpace on 2014-06-11T19:27:45Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-03-10Bitstream added on 2014-06-13T20:56:46Z : No. of bitstreams: 1 perussi_lr_me_arafo.pdf: 792637 bytes, checksum: 8a59e1121d0d7a28043ba17704db43b2 (MD5) / Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) / O laser cirúrgico de Er,Cr:YSGG (érbio, impregnado por cromo: ítrio, scandio, gálio, garnet) tem sido utilizado no tratamento cirúrgico periodontal, embora poucos estudos histológicos sejam encontrados na literatura. O objetivo deste estudo foi comparar histologicamente a utilização do laser de Er-Cr: YSGG em tecido mole e ósseo em relação à técnicas cirúrgicas convencionais. Foram utilizados 20 ratos, divididos aleatoriamente em 3, 7, 15 e 30 dias, com 5 animais por período. Em cada animal foram realizados os procedimentos (P) nas regiões de dorso e calota, respectivamente: PI) Incisão no dorso com bisturi de lâmina no 15; PII) Incisão no dorso com Laser Er,Cr:YSGG (150mJ de energia, 2.0W de potência), PIII) Defeito ósseo na calota com fresa diamantada esférica em baixa rotação sob refrigeração; PIV) Defeito ósseo na calota com Laser Er,Cr:YSGG (150mJ de energia, 3.0W de potência). Os procedimentos foram randomicamente distribuídos entre lado direito e esquerdo de cada animal. A análise histológica foi avaliada de acordo com a intensidade da resposta inflamatória, presença de áreas necróticas e processo de regeneração tecidual. A análise histométrica dos defeitos ósseos foi confirmada pelo teste estatístico paramétrico ANOVA e complementado pelo teste Bonferroni. Para os procedimentos na região de dorso pode-se observar que nos períodos iniciais houve melhor resposta histológica para o PI, embora no período de 30 dias os procedimentos I e II apresentavam resultados semelhantes. A análise histométrica confirmou uma maior formação óssea na calota com o uso do laser de Er,Cr:YSGG aos 30 dias (79.96±10.30%) em relação ao procedimento com fresa aos 15 (62.05±4.84%) e 30 dias (58.23±9.99%). Na análise histológica em tecido duro, observou-se necrose óssea superficial... / The Er,Cr:YSGG (Erbium, Chromium:Yttrium–Scandium–Gallium–Garnet) surgical laser has been demonstrated as an useful instrument on dentistry since its wavelength (2,78μm) is strongly absorbed by water and hydroxyapatite. The aim of this study was to histologically compare the in vivo healing process that occurs in soft and hard tissues incisions made by Er,Cr:YSGG laser or conventional periodontal surgeries techniques. Twenty rats were used and divided into 4 groups of 5 different periods. On each animal was performed the following procedures (P) in the soft and hard tissue: PI) dorsum incision with scalpel number 15; PII) dorsum irradiation with 2.0W Er,Cr:YSGG laser; PIII) skull defect made by a diamond spherical bur with water refrigeration; PIV) skull defect created by 3.0W Er,Cr:YSGG laser. The procedures were randomly distributed into right and left side of each animal. After sacrifice 3, 7, 15 and 30 days post-surgery the histological examinations were performed. Histometric analysis of the bone defects was confirmed by the parametric statistic test ANOVA and complemented by the Bonferroni test. In the first periods, the histological findings in the dorsum were better in the PI group, although by day 30 PI and PII showed similar healing response. The procedures in the skull revealed superficial necrosis in all periods in the PIV group, although this group had much more evidence of bone formation at day 30 than PIII. The histometric analysis also confirmed a better tissue response for laser application in the skull than conventional techniques, with 79.96±10.30% bone formation by day 30 with Er,Cr:YSGG laser and 62.05±4.84% by day 15 and 58.23±9.99% by day 30 with bur. The utilization of Er,Cr:YSGG laser in soft tissue caused a minimal delay in the healing initial periods but the... (Complete abstract, click electronic access below)

Made available in DSpace on 2014-06-11T19:23:40Z (GMT). No. of bitstreams: 0 Previous issue date: 2005-12-19Bitstream added on 2014-06-13T18:19:49Z : No. of bitstreams: 1 coutinho_ccc_me_araca.pdf: 114919 bytes, checksum: 49f558e8cba145ffdabb2e45dc62ada3 (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Na dinâmica da reparação óssea os fenômenos de reabsorção e neoformação são dependentes e acoplados. Proteínas efetivamente envolvidas na diferenciação celular determinam ativação ou inibição das atividades que regulam o ganho ou perda de massa óssea. Dentre as proteínas ósseas identificadas e envolvidas na dinâmica óssea podemos destacar a OPG, a RANK e RANKL como marcadores de atividades celulares. O presente trabalho tem como objetivo identificar, nos diferentes períodos da cronologia do processo de reparo alveolar através de técnica imunoistoquímica, a presença das proteínas OPG, RANK e RANKL. Para tanto foram utilizados 60 ratos machos submetidos à exodontia do incisivo superior direito e perfundidos aos 14, 21 e 28 dias pós-operatórios. As hemi-maxilas contendo o alvéolo dental em reparação foram removidas, pósfixadas, descalcificadas em EDTA, crioprotegidas e obtidos cortes longitudinais com 14æm em criostato. Os cortes foram submetidos à reação imunoistoquímica mediante a utilização de anticorpos primários para OPG, RANK e RANKL, como amplificador foi utilizado o sistema avidina-biotina e a diaminobenzidina (DAB) como cromógeno. Os resultados mostram que qualitativamente ocorre um balanço na expressão das proteínas que caracterizam reabsorção e neoformação óssea nos diferentes períodos estudados, onde aos 14 e 21 dias ocorre maior expressão de RANK. Com relação às proteínas OPG e RANKL, observa-se que elas apresentam-se expressas nas células da linhagem osteoblástica de forma similar, sendo que 28 dias é o período de maior expressão destas proteínas. / In the bone healing dynamics, the resorption and neoformation processes are dependent. Proteins involved in the cellular differentiation determinate the activation or inhibition of the activities that regulate the gain or loss of bone mass. From all of the identified bone proteins, it may be distinguished the OPG, RANK and RANKL. The present study has the aim to identify, in the different periods of alveolar bone healing chronology, the expression of OPG, RANK and RANKL proteins using the immunohistochemistry methodology. To perform this study, 60 male rats had the right upper incisive extracted and they were perfused at 14, 21 and 28 pos-operative days. The hemimaxilla with the rat extraction socket was removed, pos fixed and decalcified in EDTA. Then, they were cryoprotected and longitudinal slices with 14 ìm thickness were obtained in cryostat. The slices were submitted to immunohistochemistry reaction and the primary antibodies used were against OPG, RANK and RANKL proteins. It was used the avidinbiotin system to amplify the sign and diaminobenzidine was the cromogen. The results show that there is a balance in the expression of the proteins, showing that there is an increase in the expression of RANK at 14 and 21 pos-operative periods. In relation to OPG and RANKL, these proteins presents a similar expression in all of the pos-extraction periods analysed in this study and at 28 days after the extraction there is the greater expression of both proteins.

Peroxissome proliferator-activated receptor-alfa (PPAR-?) é um fator de transcrição nuclear envolvido na regulação do metabolismo de lipídeos e da inflamação. PPAR? pode estar relacionado com a modulação da cicatrização de feridas cutâneas, que é um processo multifatorial, dependente de mecanismos de sinalização celular e de inflamação. Deste modo, o objetivo deste trabalho foi analisar o papel do receptor PPAR? na cicatrização de feridas cutâneas induzidas experimentalmente e a sua relação com o metabolismo sistêmico após tratamento com agonista do PPAR?. Para tanto, foram realizadas feridas na pele da região dorsal de camundongos 129/SvEv, que foram tratados diariamente com o agonista de PPAR?, Gemfibrozil, por via oral ou tópica. Os animais foram acompanhados durante 240h pós-cirúrgico (p.c.) para a análise do reparo cutâneo e alterações metabólicas que poderiam ser induzidas pela ativação de PPAR?. Os camundongos tratados apresentaram melhor cicatrização após ativação de PPAR? com 100 ou 50 mg/kg/dia de agonista por via oral ou tópica, respectivamente. O tratamento oral induziu cicatrização mais rápida somente após 24h, 48h e 72h p.c., enquanto que os animais tratados com Gemfibrozil tópico apresentaram cicatrização mais precoce em todos os tempos avaliados. A indução de feridas alterou o metabolismo sistêmico dos camundongos que demonstraram significativa perda de peso e redução de triglicérides, independentemente do tratamento. Porém, a ativação de PPAR? não alterou a glicemia ou a função hepática. Na análise histopatológica das feridas foi verificado infiltrado inflamatório, composto principalmente por neutrófilos e outras células polimorfonucleares. Entretanto, o tratamento com Gemfibrozil tópico levou a um menor infiltrado inflamatório e diferenciada deposição de colágeno após 10 dias p.c. Além disso, houve diminuição do acúmulo de neutrófilos, macrófagos e eosinófilos quando comparados aos animais que receberam apenas o veículo. O tratamento tópico promoveu menor acúmulo de linfócitos TCD4+, TCD8+ e T??, e ainda diferenciado influxo de células dendríticas para a lesão. No entanto, não houve diferença em relação a células T reguladoras nos linfonodos drenantes, mas os animais tratados apresentaram diminuição de Foxp3 nas células CD4+CD25-. Em conclusão, PPAR? atua no reparo cutâneo, e sua ativação local acelera a cicatrização por meio da modulação da inflamação na pele. Finalmente, os resultados sugerem que PPAR? pode ser alvo importante para novas terapias que visam melhorar a cicatrização de feridas, especialmente quando ativado no local da lesão. / Peroxissome proliferator-activated receptor alpha (PPAR?) is a nuclear transcription factor involved in the regulation of lipid metabolism and inflammation. PPAR? may be associated to the modulation of wound healing, which is a multifactorial process dependent on mechanisms of cell signaling and inflammation. Then this work aimed to analyze the role of PPAR? receptor in experimental cutaneous wound healing and its relationship to the systemic metabolism of mice treated with a PPAR? agonist. For this, skin wounds were performed in the dorsal region of 129/SvEv mice, treated daily with the PPAR? agonist, Gemfibrozil, by oral or topical route. Mice were followed for 240h post-surgery (p.s.) for skin repair and metabolic changes that could be induced by PPAR? activation. There was improved wound healing in mice treated with 100 or 50 mg/Kg of PPAR? agonist by oral or topical route respectively. The oral treatment induced a better repair in the early 24h, 48h and 72h p.s. while mice treated by topical application of Gemfibrozil presented faster healing in all times evaluated. Wound\'s induction affected the systemic metabolism of mice leading to significant weight loss. PPAR? agonist did not alter glucose, triglycerides or liver function, although all injured animals had a significant decrease on triglycerides levels in the early times p.s., independent on the treatment. In histopathological examination of the wounds it was observed inflammatory infiltrate, composed mainly of neutrophils and other polymorphonuclear cells. However, topical treatment with PPAR? agonist led to lower inflammatory infiltrate and differentiated collagen deposition 10 days p.s. Furthermore, there was decrease of neutrophil, macrophages and eosinophils influx when compared to untreated mice. Topical treatment led to decrease in the TCD4+, TCD8+ e T?? lymphocytes accumulation in the lesions, and differentiated dendritic cell influx to the wounds. However there was no difference regarding CD4+CD25+ T cells in lymph nodes, but treated mice showed decrease Foxp3 expression. In conclusion, the triglycerides serum level was altered in the course of wound healing and may be associated to skin lesion, while PPAR? agonist acts in wound repair by accelerating healing and modulating neutrophil influx to the skin. Finally, our results suggested that PPAR? may be an important target for novel therapies aimed at improved wound healing, especially when administered topically.

Orientador: Marisa Masumi Beppu / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Engenharia Quimica / Made available in DSpace on 2018-08-07T04:11:53Z (GMT). No. of bitstreams: 1 Goulart_GilbertoAlessandreSoares_D.pdf: 3369983 bytes, checksum: 6b0a60782f517a23c920987f58b71dd3 (MD5) Previous issue date: 2006 / Resumo: A utilização de polímeros naturais como biomateriais tem crescido nos últimos anos. Existem polímeros naturais, como a quitosana, por exemplo, que têm grande potencial de uso nas áreas farmacêuticas, médicas, de engenharia de tecidos e engenharia biomédica (reconstituição de ossos), e matriz de liberação controlada de fármacos, com propriedades biológicas interessantes, tais como: biocompatibilidade com tecidos e órgãos humanos, não toxicidade, capacidade hemostática, bactericida, fungicida e principalmente propriedade cicatrizante. Este polímero pode ser utilizado juntamente com outros compostos, como a papaína, que é uma enzima proteolítica extraída do mamão e que possui propriedade antiinflamatória, fungicida facilita a cicatrização de tecidos epiteliais. O objetivo deste trabalho foi estudar a de utilização da quitosana e papaína no reparo de tecidos epiteliais. Para tal, foram produzidas micropartículas de quitosana com papaína sorvida em diferentes concentrações e também com a utilização de agentes reticulantes como o glutaraldeído 0,75% em massa e o tripolifosfato de sódio (TPP) 10% m/v, com a finalidade de imobilizar a enzima sobre as micropartículas de quitosana. As micropartículas foram testadas quanto a sua capacidade de liberação de papaína e avaliadas quanto ao tempo de armazenamento. Analises morfológicas (MEV), cristalográficas (DRX) e térmicas (TGA e DSC) foram realizadas para caracterização das micropartículas de quitosana, com a finalidade de verificar a influência do agente reticulante na liberação de papaína sorvida nas micropartículas de quitosana. Foi observado que as micropartículas utilizadas na forma natural com papaína sorvida sofriam deterioração com o decorrer do tempo. Por esse foi mais indicado o uso de um agente reticulante, com a finalidade de imobilizar a papaína e manter sua atividade enzimática constante. O agente reticulante escolhido foi o TPP, pelo fato de não possuir toxicidade. Foi observado que a imobilização da papaína ocorreu de forma efetiva pelo uso do TPP, pois se conseguiu manter constante a atividade enzimática nas micropartículas de quitosana reticuladas por um período de tempo de 6 meses / Abstract: The use of natural polymers as biomaterials has been growing in the last years. Natural polymers as chitosan, consists in good examples with great potential for use in pharmaceutical and medical areas, tissue and biomedical engineering (reconstitution of bones), and controlled release of drugs. Chitosan presents interesting biological properties, such as: biocompatibility with tissue and organs, non toxicicity, hemostatic, anti-bacterial, anti-fungal properties and mainly healing properties. It can be used associated with other active molecules, such as papain, that is a proteolitic enzyme of papaya and also possesses anti-inflammatory property, being able to facilitate the healing of epithelial tissues. The objective of this study was to evaluate the possibility of using of chitosan for wound healing. Hence, microparticles of chitosan with sorbed papain (in several concentrations) associated with the use of crosslinking agents as glutaraldehyde 0.75% (w/w) solution and sodium tripoly-phosphate (TPP) 10% (w/v) solution in order to immobilize the enzyme. The produced microparticles were tested on its capacity of papain release and the time of storage. Morphological (SEM), crystallographic (DRX) and thermal (TGA and DSC) analyses were performed for microparticles characterization, with the purpose to verify the influence of the crosslinking agent in the release of papain from the microparticles. As results, microparticles used in the natural form with sorbed papain underwent deterioration with elapsing of the storage time. For this reason, TPP was used to immobilize papain and keep its enzimatic activity, without presenting toxicicity. The immobilization of the papain using TPP occurred in a sactisfactory way, keeping the enzymatic activity of papain in chitosan microparticles even after a long shelf life / Doutorado / Engenharia de Processos / Doutor em Engenharia Química

Orientador: Edgard Graner / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Odontologia de Piracicaba / Made available in DSpace on 2018-07-27T09:39:40Z (GMT). No. of bitstreams: 1 Silva_HannahCarmemCarlosRibeiro_D.pdf: 2966581 bytes, checksum: 18a22acf8512cc1abc03572e8071674e (MD5) Previous issue date: 2000 / Resumo: A Ciclosporina A (CSA) é uma droga imunossupressora, que atua de forma específica e reversível, principalmente em linfócitos T CD4. O objetivo deste trabalho foi verificar o efeito da CSA sobre a produção e atividade de metaloproteinases de matriz (MMPs) associadas ao processo de reparação alveolar de molares de ratos Wistar, através de análise zimográfica. Foram utilizados 30 ratos tratados diariamente com CSA na concentração de 10 mg/kg de peso corporal por via subcutânea, com o início do tratamento ocorrendo 7 dias antes das exodontias e permanecendo após as cirurgias por períodos de 1 a 10 dias, quando os grupos foram sacrificados. Como grupo controle foram utilizados 30 ratos aplicados com solução salina a 0,9%. Verificou-se a presença de enzimas gelatinolíticas com comportamento, frente a ação de inibidores específicos e massa molecular semelhantes às das enzimas pertencentes ao grupo das MMPs. A análise nos diversos períodos de reparação alveolar demonstrou que este processo é acompanhado por secreção e atividade gelatinolítica, principalmente de MMP-9 e MMP-2. A produção e atividade gelatinolítica total e das MMP-9 e MMP-2 dos meios de cultura condicionados com o tecido de granulação dos alvéolos dos ratos foi menor nos animais tratados com CSA do que no grupo controle. Estes resultados indicam que a CSA reduz a produção e atividade de enzimas pertencentes ao grupo das MMPs, que participam no processo de reparo alveolar / Abstract: Cyclosporin A (CSA) is a potent immunosuppressive agent which acts in a specific and reversible way, mainly in T CD4 Iymphocytes. The aim of this study was to investigate the production and activity of matrix metalloproteinases (MMPs) through zymography during alveolar wound healing of inferior molars of Wistar's rats treated with CSA. Thirty Wistar rats where daily injected subcutaneously with 10 mg/kg/body weight of CSA starting seven days before the extraction and continuing from 1 to 10 days after surgery. The same numbers of rats were injected with the same volume of saline solution as a control group. Specific inhibitors were used to identify MMPs enzymes group. The data obtained from different healing periods, showed secretion and gelatinolytic activity, mainly by MMP-9 and MMP-2. The total gelatinolytic activity of MMP-9 and MMP-2 of the cell culture medium incubated with granulation tissue from the healing area, were lower in the CSA group than in the control group. The results showed that CSA use reduced the MMPs enzyme activity, which may have an important role in the alveolar wound healing process / Doutorado / Doutor em Biologia e Patologia Buco-Dental

O cuidado com as áreas doadoras de enxertos de pele merece constante atenção, visto que desencadeia desconforto por dor e restrição de movimentos. Existem evidências de que a estimulação elétrica pode acelerar a cicatrização de feridas e produzir analgesia, e que diferentes parâmetros físicos podem interferir nas respostas apresentadas. O objetivo do estudo foi avaliar o efeito da polaridade da corrente na cicatrização e na dor. Para tanto foram comparados os efeitos da estimulação elétrica de alta voltagem (EAV), polarizada, e a estimulação elétrica nervosa transcutânea (ENT), despolarizada, no tratamento das áreas doadoras de pacientes queimados. Para tanto, foram avaliados 48 voluntários do sexo masculino randomizados em três grupos: submetidos à estimulação elétrica de alta voltagem (GEAV), média idade de 34,2(±9,8) anos, n=17; submetidos à estimulação elétrica nervosa transcutânea (GENT), com 34(±9,5) anos, n= 16; e não submetido à estimulação elétrica ou grupo controle (GC) média de idade 35(±9,5 anos), n= 15. Os procedimentos terapêuticos foram aplicados nas extremidades da área doadora, no primeiro pós-operatório, até a epitelização completa. As variáveis avaliadas foram avaliação clínica, o tempo (dias) de epitelização, estimado pelo desprendimento do curativo primário sobre a lesão, avaliação da dor pela escala numérica de dor, a temperatura cutânea pela termografia infravermelha, qualidade da cicatriz (book de fotos, escala ® Vancouver, software Image J ). Após a análise dos dados, foi aplicado o teste de Shapiro-Wilk, em seguida o comportamento pré e pós-intervenção intragrupo foi aplicado o teste Wilcoxon. Para comparação entre os grupos foi efetuado teste de Kruskal-Wallis seguido de post-hoc de Dunn, em todos os casos foi utilizado o nível de significância de 5% (p<0,05). Os achados apontam que o tempo de desprendimento do curativo Rayon das áreas doadoras foi significativamente menor para GEAV apresentando (p<0,033). Houve redução significativa da dor (p<0,05) para o GEAV e para o GENT, quando comparado ao GC. A quantidade relacionada a solicitação de analgésicos foi reduzida para os grupos estimulados, com diferença significativa do GEAV versus GC (p<0,002) e GENT versus GC (p<0,001). Não houve diferença significativa na temperatura cutânea entre os grupos. Não houve diferença significativa no escore final da escala Vancouver e nem quantidade de crostas entre os grupos. A polaridade da corrente pode ter influenciado no tempo de epitelização, porém não interferiu na dor e na qualidade da área doadora. / The cares with donor areas of skin grafts deserve constant attention, since it triggers discomfort due to pain and movement restriction. There are evidences that electric stimulation may accelerate wound healing and produce pain relief. The objective of this study was to evaluate the effect of electric current polarity on the healing and pain. Therefore, the effects of stimulation on high voltage pulsed current (HVPC) and nervous transcutaneous stimulation (TENS) were compared in the treatment of donor areas of burns victims. Therefore, 48 volunteers of the male sex were randomized between three groups: submitted to high voltage pulsed current stimulation (GHVPC), with 34.2 (± 9.8) years, n=17; submitted to nervous transcutaneous stimulation (GTENS), with 34 (±9.5) years, n=16; and nonsubmitted to stimulation group, or control group (GC), average age of 35 (± 9,5) years, n=15. The therapeutic procedures were applied on the edges of the donor area, at the first postoperative, until complete healing. The evaluated variables were ephitelization time (days), estimated by the unfastening of the primary curative on the wound, evaluation of pain by numerical scale of pain, skin temperature by infrared thermography, scar quality (photo book, Vancouver scale and Image J® software). After data analysis, the Shapiro-Wilk test was applied, and the Wilcoxon test was applied to the before and after intervention. Kruskal-Wallis test followed by Dunn post-hoc was used to compare the groups. In all cases, the significance level of 5% (p<0.05) was used. The findings indicate that the time of release of the Rayon dressing from the donor sites was significantly reduced for GEAV (p <0.033). A reduction of pain relief was significant (p<0.05) for the GEAV and for the GENT, when compared to the GC. Amount the number of solitation for analgesic drugs was decreased for the groups stimulated with significant difference of the GEAV verse GC (p<0.002) and GENT verse GC (p <0.001). Change in cutaneous temperature was not significant between groups. There was no difference significant in score of Vancouver scale and in the number of crusts in the groups. The polarity of the current might have influenced the healing time, however not the pain nor the quality of the donor site.

Orientador: Joyce Maria Anicchino-Bizzacchi / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Made available in DSpace on 2018-08-22T13:12:37Z (GMT). No. of bitstreams: 1 Huber_StephanyCares_M.pdf: 5877456 bytes, checksum: 79a59ae5e01aa03f9d5a57ca56e58df7 (MD5) Previous issue date: 2013 / Resumo poderá ser visualizado no texto completo da tese digital / Abstract is available with the full electronic document / Mestrado / Clinica Medica / Mestra em Clínica Médica

Wound healing, as a normal biological process in the human body, is achieved through four precise and highly programmed phases: hemostasis, inflammation, proliferation, and remodeling. For a wound to heal successfully, all four phases must occur in the proper sequence and time frame. However, many factors can interfere with one or more of these phases, thus causing improper or impaired wound healing. Maintaining a moist wound environment is crucial in facilitating the wound-healing process. The beneficial effects of a moist versus a dry wound environment include re-epithelization, tissue granulation, and repair. The use of hydrocolloid occlusive dressings in maintaining a moist wound environment has proven to be a useful adjunct in facilitating wound healing. Although hydrocolloid dressings have been widely used clinically in wound management, bacterial resistance, poor solubility and sustained drug release remain to be a problem for many of the drugs used in wound therapy. In chronic wound management, where patients normally undergo long treatments and frequent dressing changes, a system that delivers drugs into a wound site in a controlled fashion can improve patient compliance and therapeutic outcomes. Liposomes are small phospholipid vesicles that have been widely investigated as drug carriers for the delivery of therapeutic agents. They are spherical lipid vesicles consisting of phospholipid bilayers that improve the efficacy of the drugs by fusing with biological membranes, and eventually releasing their entrapped content into the cells or bacteria. The aim of this study therefore, is to develop a new bacitracin-based controlled release hydrocolloid dressing, with good absorptive properties for improving the efficacy of antibiotics in wound healing. HPLC (high-pressure liquid chromatography) assay of bacitracin was performed for quantification of the drug. Liposomes were prepared using thin film hydration and extrusion methods. Liposomes were also characterized based on their ideal particle size and encapsulation efficiency, and then incorporated into the different ratios of chitosan/gelatin hydrocolloid films. The films were prepared with increase in gelatin concentration and were evaluated for folding endurance, tensile strength, water absorption capacity, morphology, drug release kinetics, antimicrobial activity and stability. The morphology of these films was found to be very smooth and homogeneous proving a good compatibility between the two polymers. With increase in gelatin concentration, folding endurance, water absorption capacity, tensile strength, drug release kinetics and antimicrobial activity were increased. The antibacterial activity against various bacterial species was improved in the bacitracin loaded hydrocolloid films as compared to the blank films. Based on the findings above, it can be concluded that chitosan/gelatin films at 1:3 proportion is a successful wound dressing for wound management with improved wound healing properties than other formulations. This formulation is a potential candidate for the development of alternative pharmaceutical dosage forms, for the treatment of bacterial infected wounds, based on the activity of the eco-friendly chitosan matrix added to the bacitracin activity. In this work, chitosan also demonstrated a great potential as a dressing for advanced wound therapy and confirmed its good biocompatibility and potential to provide, in combination with liposomes, sustained drug release which is highly beneficial for wound treatment. The addition of gelatin improved the water affinity of the films and facilitated water mediated cross-linking process.

Abstract Degradation of extracellular matrix (ECM) and basement membrane (BM) are required both in normal physiological conditions such as wound healing and in pathological tissue remodelling such as chronic ulcers and cancers. Matrix metalloproteinases (MMPs) are an enzyme family, which can cleave most ECM and BM components. They are associated with physiological and pathological processes but their exact roles are still largely unknown. The expression of MMP-8 and MMP-26 in acute and chronic human cutaneous wounds using histological and cell culture methods were investigated. MMP-8 was expressed in epithelial cells, neutrophils, and other inflammatory cells especially in chronic ulcers while in acute wounds MMP-8 expression was weak or absent. MMP-26 was temporarily present in acute wounds while it was strongly expressed in close vicinity to the BM in multiple cell types of most chronic ulcers. In vitro keratinocyte wound assay showed that MMP-8 and -26 were expressed in migrating cells. Bone formation, collagen metabolism, and inflammation in MMP8-/- mice tooth extraction wounds and also periapical lesion formation were analysed. No differences between wild type or MMP-8-deficient mice in the new bone area or periapical lesion size were found. However, type III procollagen production was increased and inflammatory cell influx was decreased in MMP8-/- mice. In addition, Fas ligand (FasL) production was increased in mandibular alveolar mucosa but decreased in alveolar bone of MMP-8 deficient mice. MMP-8 was also found to cleave FasL in vitro. A total of 90 human mobile tongue squamous cell carcinoma (SCC) samples were collected. Bryne’s malignancy scores, thickness of the SCCs, expression of microvessel density (CD31 and factor VIII), cyclooxygenase-2 (COX-2), the laminin-5 (currently termed laminin-332) γ2-chain, integrin αvβ6, estrogen receptor-α (ER-α), estrogen receptor-β (ER-β), and MMPs (-2, -7, -8, -9, -20, and -28) were analysed. The high expression of MMP-8 was associated with a better prognosis for the patients, particularly in females. In addition, tongue carcinoma formation in MMP8-/- mice was investigated. Tongue SCC developed more often in MMP8-/- female mice than wild type littermates. In addition, MMP-8 can cleave ER- α and -β and estrogen can induce MMP-8 production in vitro. A total of 22 biopsies, 10 resection sections, and three lung metastases of 25 osteosarcoma patients samples were stained with MMP-2, -8, -13, -26, and tissue inhibitor of metalloproteinase-1 (TIMP-1) using immunohistological methods. Expression of these markers was mostly present in sarcoma cells but MMP-8 was not present in lung metastases. In resection sections, chemotherapy altered MMP-2, -8, and -13 expressions compared to biopsies. However, an association between the expression and prognosis of osteosarcoma patients could not been found. In conclusion, MMP-8 seems to be an estrogen-related protective factor in tongue SCC and can regulate ECM and BM components and inflammation during wound healing. Further studies are needed to evaluate the exact function especially of MMP-8 in human osteosarcoma.