Thesis (Ph. D.)--West Virginia University, 2007. / Title from document title page. Document formatted into pages; contains xiii, 127 p. : ill. (some col.). Vita. Includes abstract. Includes bibliographical references.

Acceleration of the regeneration process is necessary for severe full-thickness skin injuries, such as burns, trauma or chronic ulcers. The aim of this dissertation was to investigate the influence of bone marrow-derived stem cells on skin regeneration using full-thickness wound model in BALB/c mouse. Lin¯ cells were isolated and identified. Regenerative properties of purified cells were evaluated in vitro. Wound healing after Lin¯ cells transplantation was examined histologically and cytokine gene expression was determined by quantitative RT-PCR. The results revealed that Lin¯ population is heterogeneous and contains distinct undifferentiated cell types. In vitro experiments showed that skin tissue extracellular matrix components enhance proliferation and migration of Lin¯ cells. Histologic analysis indicated that inhibition of inflammation, re-epithelization, formation of skin appendages and extracellular collagen, were most effective after cell transplantation with type I collagen. Quantitative analysis revealed that transplanted Lin¯ cells decrease expression of proinflammatory cytokine TNF-α and increase anti-inflammatory IL-10 expression. Also, gene expression of growth factors TGF-β and VEGF, which enhance regeneration, was improved in different healing phases. The results obtained provide additional knowledge for possible therapeutic application of stem/progenitor cells in difficult-to-heal wounds. / Odos žaizdų regeneracijos skatinimas yra aktualus pažeidus didelį paviršiaus plotą bei giliuosius audinio sluoksnius. Šio darbo tikslas buvo ištirti kaulų čiulpų Lin¯ ląstelių įtaką odos regeneracijai in vivo, taikant BALB/c linijos pelių visų odos sluoksnių pažeidimo modelį. Darbo metu buvo išskirtos bei identifikuotos Lin¯ ląstelės. Jų regeneracinės savybės buvo įvertintos in vitro. Buvo atlikta histologinė gyjančio audinio analizė bei nustatyti citokinų genų raiškos pokyčiai. Rezultatai parodė, kad Lin¯ ląstelių populiacija yra heterogeniška, sudaryta iš nediferencijuotų ląstelių. Nustatyta, kad odos tarpląstelinės medžiagos komponentai skatina Lin¯ ląstelių proliferaciją ir migraciją in vitro. Histologinė analizė parodė, kad po ląstelių transplantacijos I tipo kolageno gelyje anksčiausiai prasideda uždegimo slopinimas, greičiausiai baigiamas reepitelizacijos procesas, atsinaujinusioje dermoje susiformuoja tvarkingai išsidėsčiusios pagalbinės odos struktūros bei natyviam audiniui būdingas tarpląstelinės medžiagos kolagenas. Atlikus kiekybinę analizę nustatyta, kad po Lin¯ ląstelių transplantacijos sumažėja uždegiminio citokino TNF-α raiška bei padidėja priešuždegiminio IL-10 raiška. Skirtingose gijimo stadijose padidėja augimo veiksnių, skatinančių audinio regeneraciją, TGF-β ir VEGF genų raiška. Gauti rezultatai papildo žinias, reikalingas vystyti metodus, skirtus sutrikusio gijimo žaizdų regeneracijos skatinimui.

Thesis (Ph. D.)--Ohio State University, 2006. / Title from first page of PDF file. Includes bibliographical references (p. 214-230).

Submitted by (edna.saturno@ufrpe.br) on 2016-10-11T13:20:49Z No. of bitstreams: 1 Flavio de Oliveira Silva.pdf: 672464 bytes, checksum: 70191679dda58a1314e6dce99c1b08da (MD5) / Made available in DSpace on 2016-10-11T13:20:49Z (GMT). No. of bitstreams: 1 Flavio de Oliveira Silva.pdf: 672464 bytes, checksum: 70191679dda58a1314e6dce99c1b08da (MD5) Previous issue date: 2007-02-05 / Lectins, carbohydrate binding proteins of non immune origin, agglutinate cells and glycoconjugates. Some proteins as the lectins from the Diocleinae tribe have described as pharmacological active molecules. Lectins form an important class of carbohydrate-binding proteins. Canavalia brasiliensis (ConBr) is a sugar-binding specificity [D-mannose (Dglucose)] protein. The aim of this study was evaluate the proteins profile and to evaluate histhopathologically treated wounds with ConBr seeds lectins. Surgical wounds were produced in the dorsal region in albino Swiss mice. Each wound was daily treated topically along 11 days, as follow: ConBr (10μg), ConBr/Manose (10μg in manose 0,1M), manose (0,1M) and NaCl (150mM). Protein profile has done through SDS-PAGE and showed proteins with molecular weight between 30 and 70KDa, suggesting the presence of metalloproteinases in the homogenates gotten from the cutaneous injuries. In 2nd day of postoperative (PO), the ConBr/Manose group was what it presented the highest proteolitic activity, in 7th day of P.O., the highest activity was observed in the ConBr and NaCl groups. Finally, in 12th day of PO ConBr/manose group showed a maxim proteolitic activity. ConBr group showed flogistic signs less intense than others groups, had presented contraction percentile superior during the fibroplasia phase. In the first 48 hours it was observed, in all the studied groups, the presence of polymorphonuclear leukocyte infiltrate, crust and angiogenesis. At 7th day there was presence of fibrovascular (manose, ConBr Manose and ConBr) and vascular granulation tissue (NaCl) in studied groups. Finally, at 12th day, wounds showed reepithelization. In ConBr/manose group there was presence of fibrovascular granulation tissue with more organized collagen network. Results suggests that wound healing it was better in ConBr group, having presented one better effect when used in association with manose. / Lectinas são proteínas de origem não imune que se ligam a carboidratos, podendo aglutinar células e glicoconjugados. Algumas dessas lectinas, como as obtidas de leguminosas da tribo Diocleinae possuem atividade farmacológica, a exemplo da Canavalia brasiliensis (ConBr) a qual exibe especificidade de ligação D-manose/D-glicose. O objetivo desse estudo foi determinar o perfil de proteases e avaliar clínica e histopalogicamente o efeito do tratamento tópico de feridas cutâneas com a ConBr livre e conjugada com o seu açúcar específico. Para isso, lesões cirúrgicas foram produzidas na região dorsal de camundongos, divididos em grupos, de acordo com o tratamento que se segue: NaCl (150mM), manose (0,1M), ConBr (10μg) e ConBr/manose (10μg em solução fisiológioca com manose 0,1M). O perfil das proteínas realizado através de eletroforese SDS-PAGE demonstrou a presença de proteínas com peso molecular compreendidos na faixa de 30 a 67 kDa, sugerindo a presença de metaloproteinases nos homogenatos obtidos a partir das lesões cutâneas. No 2º dia de pósoperatório (P.O.), o grupo ConBr/Manose foi o que apresentou a maior atividade proteolítica, no 7º dia de P.O., a maior atividade foi observada nos grupos ConBr e NaCl, finalmente, no 12º dia de P.O. a maior atividade proteolítica foi observada no grupo ConBr/manose. Na avaliação clínica observou-se que o grupo ConBr apresentou sinais inflamatórios menos intensos que os outros grupos avaliados, tendo evoluído com maior percentual de contração durante a fase de fibroplasia. Nas análises histopatógicas visualizou-se no 2º dia de P.O., a presença de crosta, infitrado polimorfonuclear e neoangiogênese em todos os grupos estudados. No 7º dia de P.O., os grupos Manose, ConBr/Manose e ConBr apresentaram tecido de granulação com padrão fibrovascular enquanto no grupo NaCl foi visualizado tecido de granulação com padrão vascular. No 12º dia todos os grupos apresentaram reepitelização, porém o grupo ConBr/Manose apresentou melhor padrão de organização das fibras colágenas. A lectina isolada das sementes da Canavalia brasiliensis favoreceu o processo cicatricial das lesões cutâneas experimentais, tendo apresentado um melhor efeito sob o ponto de vista histopatológico quando usada em associação com a manose.

The diploma thesis if focused on the study of bioactive hydrogél and nanofiber wound dressings composed of natural biopolymers, which were functionalized by active compounds in the form of analgesic, antibiotics and enzymes. Hydrogél wound dressings were constituted from alginate and chitosan and nanofibers were created from polyhydroxybutyrate. The following 7 active compounds were selected to be added to the wound dressings: ampicillin, streptomycin, ibuprofen, papain, bromelain, collagenase and trypsin. In the theoretical part the structure of the skin and types of wound injuries were described. This part also talks about types of wound dressing and their applications, as well as treatment of skin wounds using enzymes and compounds with analgesic and antimicrobial properties. In addition, this section describes safety assays, in particular cytotoxicity assays on human cells. At the beginning of the experimental part, the process of preparation of hydrogél wound dressing was optimised. Subsequently, the dressings were enriched with active compounds and the rate of gradual releasing of the substances into model environment was monitored. In the case of enzymes, their proteolytic activity was also tested after their incorporation to the wound dressings. Furthermore, the prepared bioactive wound dressings were analyzed for possible cytotoxic effect on human keratinocytes. Finally, the wound dressing with combined content of active substances was created and also characterized for the rate of substance release, proteolytic activity and cytotoxicity. Antimicrobial activity of this wound dressings, against two selected strains of microorganisms: Escherichia coli and Staphylococcus epidermidis, was also evaluated.

京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第17976号 / 医博第3840号 / 新制||医||1001(附属図書館) / 80820 / 京都大学大学院医学研究科医学専攻 / (主査)教授 戸口田 淳也, 教授 宮地 良樹, 教授 長澤 丘司 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DGAM

Made available in DSpace on 2016-01-26T18:55:31Z (GMT). No. of bitstreams: 1 MILENA dissertacao_12_08_2011.pdf: 499305 bytes, checksum: 3ca75cd129170949ca78544d58d71285 (MD5) Previous issue date: 2011-06-20 / Wound healing aims to restore the morphological and functional integrity of the skin. This study evaluates by means of clinical and histologic healing process of skin wounds induced experimentally in rats using a film crystalline cellulose Veloderm ®. Thirty-two rats were divided into two groups: control group (CG) wounds treated with a solution of sodium chloride 0.9% and group veloderm (GV) wounds treated with a film of crystalline cellulose and were evaluated for 26 days at different times. Weight loss was observed in animals from both groups in the early stages, and greater weight gain in the final moments of the GV to the animal, the temperature oscillations in the two groups with predominance in some moments of hypothermia, pinkish wound in the two groups across all time points, greater granulation tissue in animals of CG, the presence of little oozing from the wound and feature in GV and more serous exudation in characteristic bloody GC, GC in the presence of pain and pain in the absence of GV and greater contraction of the wound to the GC, but with complete healing in early GV. Thus, we conclude that the crystalline cellulose film Veloderm ® is effective in the treatment of skin wounds in rats, easy to apply and use, promotes protection and lessen the pain by bringing comfort to the patient, enhances visualization and control of the evolution of the injury keeping humidity, as well as a cost-effective. / A cicatrização de feridas visa restabelecer a integridade morfológica e funcional da pele. Este estudo avalia por meio de exame clínico e histológico o processo de cicatrização de feridas cutâneas induzidas experimentalmente em ratos Wistar, utilizando uma película de celulose cristalina denominada Veloderm®. Trinta e dois ratos foram distribuídos em dois grupos: grupo controle (GC) feridas tratadas com solução de cloreto de sódio 0,9% e grupo veloderm (GV) feridas tratadas com a película de celulose cristalina e foram avaliados durante 26 dias em diferentes momentos. Foi observado perda de peso nos animais dos dois grupos nos momentos iniciais, e maior peso nos momentos finais para os animais do GV, oscilações da temperatura nos dois grupos com predominância em alguns momentos de hipotermia, coloração rósea da ferida nos dois grupos ao longo de todos os momentos de avaliação, maior tecido de granulação nos animais do GC, presença de pouca exsudação da ferida e de característica serosa no GV e maior exsudação de característica sanguinolenta no GC, presença de dor no GC e ausência de dor no GV e maior contração da ferida para o GC, porém com cicatrização completa da ferida mais precoce no GV. Desta forma, conclui-se que a película de celulose cristalina Veloderm® é eficaz no tratamento de feridas cutâneas em rato, de fácil aplicação e utilização, promove proteção e diminui a dor trazendo conforto ao paciente, favorece visualização e controle evolutivo da lesão mantendo a umidade, além de um bom custo benefício.

Made available in DSpace on 2016-07-18T17:53:08Z (GMT). No. of bitstreams: 1 MILENA dissertacao_12_08_2011.pdf: 499305 bytes, checksum: 3ca75cd129170949ca78544d58d71285 (MD5) Previous issue date: 2011-06-20 / Wound healing aims to restore the morphological and functional integrity of the skin. This study evaluates by means of clinical and histologic healing process of skin wounds induced experimentally in rats using a film crystalline cellulose Veloderm ®. Thirty-two rats were divided into two groups: control group (CG) wounds treated with a solution of sodium chloride 0.9% and group veloderm (GV) wounds treated with a film of crystalline cellulose and were evaluated for 26 days at different times. Weight loss was observed in animals from both groups in the early stages, and greater weight gain in the final moments of the GV to the animal, the temperature oscillations in the two groups with predominance in some moments of hypothermia, pinkish wound in the two groups across all time points, greater granulation tissue in animals of CG, the presence of little oozing from the wound and feature in GV and more serous exudation in characteristic bloody GC, GC in the presence of pain and pain in the absence of GV and greater contraction of the wound to the GC, but with complete healing in early GV. Thus, we conclude that the crystalline cellulose film Veloderm ® is effective in the treatment of skin wounds in rats, easy to apply and use, promotes protection and lessen the pain by bringing comfort to the patient, enhances visualization and control of the evolution of the injury keeping humidity, as well as a cost-effective. / A cicatrização de feridas visa restabelecer a integridade morfológica e funcional da pele. Este estudo avalia por meio de exame clínico e histológico o processo de cicatrização de feridas cutâneas induzidas experimentalmente em ratos Wistar, utilizando uma película de celulose cristalina denominada Veloderm®. Trinta e dois ratos foram distribuídos em dois grupos: grupo controle (GC) feridas tratadas com solução de cloreto de sódio 0,9% e grupo veloderm (GV) feridas tratadas com a película de celulose cristalina e foram avaliados durante 26 dias em diferentes momentos. Foi observado perda de peso nos animais dos dois grupos nos momentos iniciais, e maior peso nos momentos finais para os animais do GV, oscilações da temperatura nos dois grupos com predominância em alguns momentos de hipotermia, coloração rósea da ferida nos dois grupos ao longo de todos os momentos de avaliação, maior tecido de granulação nos animais do GC, presença de pouca exsudação da ferida e de característica serosa no GV e maior exsudação de característica sanguinolenta no GC, presença de dor no GC e ausência de dor no GV e maior contração da ferida para o GC, porém com cicatrização completa da ferida mais precoce no GV. Desta forma, conclui-se que a película de celulose cristalina Veloderm® é eficaz no tratamento de feridas cutâneas em rato, de fácil aplicação e utilização, promove proteção e diminui a dor trazendo conforto ao paciente, favorece visualização e controle evolutivo da lesão mantendo a umidade, além de um bom custo benefício.

The diploma thesis is focused on the study of fibrous wound dressings prepared by electrospinning method from natural biopolymers. Three active ingredients were added to the dressings: ampicillin, ibuprofen and collagenase, which are responsible for relieving pain, reducing the risk of infection and selectively removing necrotic tissue in the wound. The theoretical part describes the therapeutic dressings currently available on the market and the most common methods of nanofiber production. The experimental part evaluates the optimization of the preparation of gelatin, alginate and chitosan fibrous wound dressings, which were subsequently enriched with active substances and their gradual release into the model environment was determined spectrophotometrically. Antimicrobial effects against E.coli and S. epidermidis strains andantifungal activity against C. glabrata yeast were monitored. Finally, two cytotoxicity tests on the human keratinocyte cell line HaCaT confirmed the safety of the prepared products, which can serve as bioactive skin dressings in the future.

The presented diploma thesis deals with the optimalisation of preparation and the preparation of combined nanostructured antimicrobial biomaterials itself. In the theoretical part, a review focused on used materials and consequently preparation of nanoparticles and nanofibers was elaborated. Furthermore, the used antimicrobial substances – curcumin and ampicillin, and the principle of cytotoxicity assay were described.In practical part the optimalisation process is described. Furthermore the safety of prepared materials and used antimicrobial substances on HaCaT cell line was tested, in order to confirm their possible further use in cosmetic and pharmaceutical industry. Great part of the thesis deals with evaluation of the antimicrobial activity of used substances and prepared combined nanomaterials on multiple microorganisms from grampositive bacteria, gramnegative bacteria and yeasts. Also the release speed of antimicrobial substances from prepared nanomaterials was determined by spectrophotometer. The amount of released ampicillin from prepared nanomaterials was determined by liquid chromatography.