Thesis (doctoral)--Lunds Universitet, Malmö, 1998. / Added t.p. and errata sheet inserted. Includes bibliographical references.

Thesis (doctoral)--Lunds Universitet, Malmö, 1998. / Added t.p. and errata sheet inserted. Includes bibliographical references.

Promoting tissue repair in adults represents a major unmet medical need. Tissue regeneration relies on the activation of quiescent endogenous stem cells and whilst considerable progress has been made, novel therapies based on stem cells have thus far failed to translate to routine clinical practice. Much attention has focussed on administrating exogenous stem cells expanded or altered in vitro. However, this approach faces several major limitations, including poor engraftment, high cost and importantly, the necessity for immunosuppression for allogenic cells. An alternative strategy that overcomes these shortcomings is to promote repair by harnessing the regenerative potential of endogenous stem cells. This thesis focussed on the role of alarmins, the upstream group of mediators released immediately following injury, in promoting tissue regeneration. I show that the alarmin HMGB1 accelerates tissue regeneration in various systems by transitioning multiple stem cell types from the quiescent G₀ to the G_Alert state. Firstly, in vitro screening of candidate alarmins with human bone marrow-derived mesenchymal stromal cells showed that only pre-treatment with HMGB1 improved osteogenic differentiation. Using an optimised murine fracture model, I found that local administration of HMGB1 accelerated bone regeneration via the CXCL12-CXCR4 axis, whilst healing was impaired in inducible Hmgb1^-/- animals. Cell cycle analyses revealed that HMGB1 transitioned the skeletal stem cell from G₀ to G_Alert. In this intermediate phase, the cell is more metabolically active and efficiently enters the cell cycle when exposed to activating factors. This effect also extended to other types of stem cells, including murine haematopoietic and muscle stem cells, as well as human haematopoietic stem and progenitor cells, and MSCs. Using murine models, HMGB1 accelerated recovery from injury to these tissues. Finally, treatment with a single systemic dose of HMGB1 two weeks before injury also accelerated bone, haematopoietic, and muscle regeneration, which was indicative of an acquired pro-regenerative signature. These findings document that the HMGB1-G_Alert pathway results in a dynamic and adaptive multi-tissue regenerative response, and suggest that exogenous HMGB1 may be of therapeutic benefit in diverse clinical contexts, including trauma, chemotherapy, and elective surgery.

Made available in DSpace on 2014-06-11T19:31:07Z (GMT). No. of bitstreams: 0 Previous issue date: 2011-08-12Bitstream added on 2014-06-13T19:47:14Z : No. of bitstreams: 1 borrasca_ag_dr_araca.pdf: 844710 bytes, checksum: d1c813312e1b09f119604bb41d9ee6ab (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Proposição: O objetivo deste trabalho foi avaliar, qualitativa e quantitativamente, o comportamento da membrana de matriz óssea homógena desmineralizada e conservada em glicerinae e membrana de Ptfe no processo de reparo ósseo de cavidades cirúrgicas preenchidas com osso autógeno realizadas em tíbias de ratos Material e Métodos: Foram utilizados 60 ratos (Rattus norvegicus albinus, Wistar) machos, com aproximadamente 250 gramas divididos em três grupos: o grupo I (Controle-sem membrana), grupo II (membrana homógena) e grupo III (membrana de ptfe). Após tricotomia, anestesia e antissepsia foi realizada uma incisão longitudinal nas regiões antero-laterais de cada tibia. Após exposição das tíbias, duas cavidades cirúrgicas de 2mm de diâmetro foram preparadas com trefina em baixa-rotação refrigerada, uma no membro posterior esquerdo e outra no membro posterior direito, sendo a primeira preenchida com o osso autógeno particulado coletado durante a realização das duas cavidades e recobertas pelas membranas utilizadas no estudo. Aos 10 e 60 dias pós-operatórios os animais foram eutanaziados e as peças obtidas foram processadas laboratorialmente para a realização de cortes semi-seriados com seis micrômetros de espessura e corados pela hematoxilina e eosina para análise histomorfométrica. Resultados e Conclusão: A membrana homógena mostrou-se biocompatível e cumpriu sua função e em termos de área óssea neoformada as membranas obtiveram desempenho semelhantes / Purpose: The aim of this study was to evaluate the behavior of membrane of a demineralized homogenous bone matrix and preserved in glycerin and the Ptfe membrane in the process of bony repair of surgical cavities filled out with autogenous bone accomplished in tibias rats. Materials and Methods: Sixty male rats weighing approximately 250 g were selected and assigned to two groups, as follows. Group I (without membrane), Group II (homogenous membrane) and Group III ( Ptfe membrane). After anesthesia, shaving and antisepsis, a longitudinal incision was made in the anterolateral areas of each tibia. After exhibition of the tibias, two surgical cavities of 2mm of diameter were prepared with trefina in refrigerated low-rotation, one in the left posterior member, other in the right subsequent member, being the first filled out with particulate autogenous bone collected during the accomplishment of the two cavities and covered with the membrane. Ten and sixty days postoperatively, the animals were euthanized and the anatomical pieces were submitted to routine laboratorial processing and serially sectioned to obtain 6-μm-thick sections, which were stained by hematoxylin and eosin for histomorphometric analysis. Results and Conclusions: The homogenous membrane proved to be biocompatible and fulfilled its function and in terms of bone area formation membranes had similar performance

Orientador: Osvaldo Magro Filho / Coorientador: Alessandra Marcondes Aranega / Banca: Edmur Aparecido Callestini / Banca: Eleonor Álvaro Garbin Júnior / Banca: Daniela Ponzoni / Banca: Celso Koogi Sonoda / Resumo: Proposição: O objetivo deste trabalho foi avaliar, qualitativa e quantitativamente, o comportamento da membrana de matriz óssea homógena desmineralizada e conservada em glicerinae e membrana de Ptfe no processo de reparo ósseo de cavidades cirúrgicas preenchidas com osso autógeno realizadas em tíbias de ratos Material e Métodos: Foram utilizados 60 ratos (Rattus norvegicus albinus, Wistar) machos, com aproximadamente 250 gramas divididos em três grupos: o grupo I (Controle-sem membrana), grupo II (membrana homógena) e grupo III (membrana de ptfe). Após tricotomia, anestesia e antissepsia foi realizada uma incisão longitudinal nas regiões antero-laterais de cada tibia. Após exposição das tíbias, duas cavidades cirúrgicas de 2mm de diâmetro foram preparadas com trefina em baixa-rotação refrigerada, uma no membro posterior esquerdo e outra no membro posterior direito, sendo a primeira preenchida com o osso autógeno particulado coletado durante a realização das duas cavidades e recobertas pelas membranas utilizadas no estudo. Aos 10 e 60 dias pós-operatórios os animais foram eutanaziados e as peças obtidas foram processadas laboratorialmente para a realização de cortes semi-seriados com seis micrômetros de espessura e corados pela hematoxilina e eosina para análise histomorfométrica. Resultados e Conclusão: A membrana homógena mostrou-se biocompatível e cumpriu sua função e em termos de área óssea neoformada as membranas obtiveram desempenho semelhantes / Abstract: Purpose: The aim of this study was to evaluate the behavior of membrane of a demineralized homogenous bone matrix and preserved in glycerin and the Ptfe membrane in the process of bony repair of surgical cavities filled out with autogenous bone accomplished in tibias rats. Materials and Methods: Sixty male rats weighing approximately 250 g were selected and assigned to two groups, as follows. Group I (without membrane), Group II (homogenous membrane) and Group III ( Ptfe membrane). After anesthesia, shaving and antisepsis, a longitudinal incision was made in the anterolateral areas of each tibia. After exhibition of the tibias, two surgical cavities of 2mm of diameter were prepared with trefina in refrigerated low-rotation, one in the left posterior member, other in the right subsequent member, being the first filled out with particulate autogenous bone collected during the accomplishment of the two cavities and covered with the membrane. Ten and sixty days postoperatively, the animals were euthanized and the anatomical pieces were submitted to routine laboratorial processing and serially sectioned to obtain 6-μm-thick sections, which were stained by hematoxylin and eosin for histomorphometric analysis. Results and Conclusions: The homogenous membrane proved to be biocompatible and fulfilled its function and in terms of bone area formation membranes had similar performance / Doutor

Made available in DSpace on 2014-06-11T19:28:03Z (GMT). No. of bitstreams: 0 Previous issue date: 2002-03-01Bitstream added on 2014-06-13T18:07:24Z : No. of bitstreams: 1 silverio_kg_me_arafo.pdf: 1644294 bytes, checksum: 5d943591d5c51a3846c75163db9a2ebb (MD5) / O objetivo do presente estudo foi avaliar in vitro o efeito do condicionamento radicular com fator de crescimento de fibroblastos básico (b-FGF) sobre a morfologia e densidade de fibroblastos. Para tal, blocos de dentina com 4 mm2 de área de superfície foram obtidos de raízes de dentes humanos extraídos devido severo envolvimento periodontal, sendo instrumentados manualmente e autoclavados. Noventa amostras foram selecionadas aleatoriamente e distribuídas em 3 grupos segundo o tratamento de superfície prévio ao condicionamento com o b-FGF: sem tratamento - controle; 50 mg/mL de cloridrato de tetraciclina e EDTA a 24%. As 30 amostras de cada um destes 3 grupos foram distribuídas em 3 subgrupos quanto à dose de b-FGF: 0 æg/mL - controle; 50 æg/mL e 125 æg/mL. Após os tratamentos, as amostras foram incubadas a 37º C e 98% de umidade com 2mL de meio Eagle, sendo 1mL com fibroblastos de linhagem contínua (células McCoy) na concentração de 1 x 105 células/mL e 1mL meio sem células, por 24 h. Após as 24 h, as amostras foram submetidas a preparo de rotina para MEV e então, fotomicrografadas nos aumentos de 500X (densidade celular) e 1000X (morfologia celular). Em seguida, as fotomicrografias foram avaliadas por 3 examinadores treinados, calibrados, independentes e cegos, os quais verificaram morfologia e densidade celular segundo os escores propostos por Gamal et al. (1998) e Jenkins et al. (1988), respectivamente. A aplicação da Análise de Regressão pela Técnica da Árvore demonstrou haver diferenças estatisticamente significantes para a densidade celular (p<0,0001) entre os grupos EDTA, tetraciclina e controle, sendo que também houve diferenças entre as doses de 0/50 æg e 125 æg de b-FGF nas amostras condicionadas com EDTA (p<0,0001) e entre as doses de 0 e 50/125 æg de b-FGF nas amostras condicionadas com tetraciclina... . / The aim of this study was to evaluate in vitro the effect of the root surface conditioning with basic fibroblast growth factor (b-FGF) about morphology and density of fibroblasts. Dentin slices of with 4 mm2 of surface area were obtained from roots of teeth extracted due to severe periodontal involvment. These were scaled and sterilized. Ninety samples were randomly distributed into 3 groups according to treatment before application of b-FGF: non-treated - control; 50mg/mL of tetracycline HCl and EDTA 24%. The thirty samples of each group were distributed into 3 subgroups according to the concentration of b-FGF: 0 æg/mL - control; 50 æg/mL and 125 æg/mL. After treatments, the samples were incubated at 37ºC and 98% humidity with 1mL of Eagle Medium with 1 x 105 cells/mL of fibroblast from continuos lineage (McCoy Cells) plus 1mL this solution without cells during 24 hours. The samples were submitted to routine preparation for SEM and photographed at 500x (density celular) and 1000x (morphology celular). Three independent and blind examiners evaluated the fibroblast`s morphology and density, according to Gamam et al. (1998) and Jenkins et al. (1998), respectively. Classification and Regression Trees test results indicated significant differences on the density (p<0,0001) among EDTA, tetracycline and control groups with also differences between concentrations of 0/50æg and 125æg of b-FGF at the samples conditioning with EDTA (p<0,0001) and between concentrations of 0 and 50/125 æg of b-FGF at the samples conditioning with tetracycline(p<0,0001). The results of this test to morphology indicated significant differences between treatment or non-treatment with b-FGF, and that concentration of 125 æg demonstrated to be more favorable than the concentration of 50 æg. In conclusion, the treatment of root surfaces with b-FGF influenced the density... (Complete abstract, click electronic address below).

O estudo de biomateriais aplicados à cicatrização levou à modalidade do tratamento chamada de regeneração tecidual guiada (RTG). A RTG reconstitui novos tecidos usando membranas como barreira de proteção da área defeituosa, impedindo a invasão de outros tecidos, especialmente dos tecidos conjuntivos fibrosos. Em outras palavras, a RTG é uma forma de isolar o tecido ósseo do tecido mole, resultando dessa forma em uma ótima condição para a formação óssea. Atualmente, mais de 20 tipos diferentes de polímeros são usados em aplicações biomédicas, e entre eles está o colágeno, polímero ao qual, pela sua hidrólise parcial pode-se obter a gelatina. A gelatina é um polipeptídeo biodegradável, biocompatível, que apresenta anti-genicidade, plasticidade e elasticidade. A prata tem seu efeito antimicrobiano bem conhecido, com utilidade em diferentes campos da medicina, odontologia e biomateriais. Nesse contexto, este trabalho desenvolve e estuda o comportamento das membranas de gelatina e membranas de gelatina com prata para aplicações em RTG. As Membranas de gelatina foram obtidas com concentração de 4% em massa e as membranas de gelatina com prata foram obtidas nas concentrações de 0,01%, 0,05% e 0,1% da solução. Na etapa de reticulação, as membranas secas foram imersas em solução tampão de fosfato de sódio com concentrações de glutaraldeído (GTA) a 0,5%, 1% e 2%, 24 horas. O ensaio de intumescimento foi realizado em membranas reticuladas e não reticuladas, pesadas e imersas em solução tampão de fosfato pH 7,4 por períodos de 5 minutos, 15 minutos, 30 minutos, 1 hora, 3 horas, 22 horas e 24 horas e durante 8 semanas. Obteve-se membranas de gelatina com prata mediante a metodologia de adição da solução de Ag compravada pelas técnicas de DRX, FTIR, MEV e pelo teste de halo de inibição. As membranas reticuladas apresentaram menor valor de porcentagem de intumescimento quanto maior foi a concentração do agente reticulante, com exceção das amostras com concentração de 0,1M de prata. Além da atuação do GTA a prata também interfere no processo de intumescimento, aspecto e comportamento das membranas de gelatina. O efeito bactericida foi avaliado utilizando ensaio de cultura de bactérias - Teste de halo de inibição, onde as membranas de gelatina com Ag apresentaram efeito bacteriostático contra bactéria Gram positiva e Gram negativa. / The study of biomaterials applied to healing led to the treatment modality called guided tissue regeneration (GTR). The RTG reconstruct new tissue using the membranes as a barrier to protect the defective area, preventing the invasion of other tissues, especially connective tissue fibers. In other words, the RTG is a way of isolating the bone tissue of the soft tissue, thereby resulting in an excellent condition for bone formation. Currently, more than 20 different types of polymers are used in biomedical applications, and among them is the collagen polymer to which, by its partial hydrolysis can get the gelatin. Gelatin is a polypeptide biodegradable, which has antigenicity, plasticity and elasticity. Silver has antimicrobial effect well known, are useful in different fields of medicine, dentistry and biomaterials. In this context, this paper develops and studies the behavior of membranes of gelatin and gelatin silver membranes for applications in RTG. Membranes were obtained with gelatin concentration of 4% by weight and gelatin silver membranes were obtained at concentrations of 0.01%, 0.05% and 0.1% of the solution. In the crosslinking step, the dried membranes were immersed in sodium phosphate buffer to concentrations of glutaraldehyde (GTA) at 0.5%, 1% and 2% 24 hours. The swelling test was performed on crosslinked and uncrosslinked membranes, weighed and immersed in phosphate buffer pH 7.4 for a period of 5 minutes, 15 minutes, 30 minutes, 1 hour, 3 hours 22 hours and 24 hours, and for 8 weeks. Obtained gelatin membranes with silver by the method of adding the solution of Ag proven by XRD techniques, FTIR, SEM and the zone of inhibition test. The crosslinked membranes had lower percentage of swelling the higher the concentration of crosslinking agent, except for the samples with a concentration of 0.1 M silver. In addition to operating the GTA silver also interferes with the process of swelling, appearance and behavior of gelatin membranes. The bactericidal effect was evaluated by bacterial culture test - Test of inhibition zone, where the gelatin membrane with Ag Ag exhibited bacteriostatic effect exhibited against Gram positive and Gram negative.

Orientador: Enilson Antonio Sallum / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Odontologia de Piracicaba / Made available in DSpace on 2018-08-26T23:34:09Z (GMT). No. of bitstreams: 1 Franca_IsabelaLima_M.pdf: 1330190 bytes, checksum: 6782efacdca31e69f907222817356c7c (MD5) Previous issue date: 2015 / Resumo: O objetivo deste estudo foi avaliar, clinicamente, a utilização do Retalho Semilunar Posicionado Coronariamente (RSPC) para tratamento de recessões gengivais, com ou sem associação à proteína derivada da matriz do esmalte (EMD). Foram selecionados 30 pacientes, que foram randomizados e alocados em dois grupos: teste (RSPC + EMD) e controle (RSPC sozinho). Para serem incluídos no estudo, os indivíduos deveriam apresentar recessões gengivais vestibulares localizadas classe I de Miller com altura maior ou igual a 2,0mm e menor que 4,0 mm, em caninos ou pré-molares superiores. Parâmetros clínicos avaliados: altura da recessão gengival (ARG), largura da recessão gengival (LRG), nível de inserção clínica (NIC), profundidade de sondagem (PS), altura de tecido queratinizado (ATQ), espessura de tecido queratinizado (ETQ) e altura (AP) e largura (LP) das papilas mesial e distal, além de índice de placa (IPL) e índice gengival (IG). Estes parâmetros foram medidos nos seguintes períodos: baseline, 90 dias e 180 dias após o procedimento cirúrgico. Nenhuma diferença estatisticamente significante foi observada entre os grupos em relação à redução da retração gengival com 6 meses de acompanhamento, embora tenha sido encontrada maior porcentagem de cobertura radicular no grupo RSPC+EMD (91%), quando comparado ao RSPC (87%) (p>0,05). Cobertura radicular completa foi obtida em 60% dos sítios no RSPC enquanto no grupo RSPC+EMD foi observada em 66,67% dos sítios. Dentro dos limites do presente estudo pôde-se concluir que o RSPC, associado ou não a EMD, levou a redução da recessão gengival, sem diferença estatística entre os grupos, após 6 meses de acompanhamento pós-operatório / Abstract: The aim of this study was to evaluate clinically the use of the Semilunar Coronally Positioned Flap (SCPF) for the treatment of gingival recessions, with or without Enamel Matrix Derivative (EMD). Thirty patients were selected, randomized and allocated in two groups: test (SCPF + EMD) and control (SCPF alone). To attend the study, subjects should present buccal Miller class I gingival recessions with height greater than or equal to 2.0 mm and less than to 4.0 mm in maxillary canines or premolars. Clinical parameters evaluated: gingival recession height (GRH), gingival recession width (GRW), clinical attachment level (CAL), probing depth (PD), height (HKT) and thickness (TKT) of keratinized tissue and papillas height (HP) and width (LP), as well as plaque and gingival index. These data were collected at baseline, 90 days and 180 days after surgery. No statistically significant difference was observed between the groups regarding the reduction of gingival recession after 6 months of follow-up, although a higher percentage of root coverage was found in SCPF + EMD group (91%), when compared to the SCPF (87%) (p> 0.05). Complete root coverage was observed in 60% of the sites of the control group (SCPF alone) and in 66,67% of the sites of the test group (SCPF+EMD). Within the limits of this study it was concluded that SCPF, associated or not with EMD may provide a reduction in gingival recession, with no statistical difference between groups / Mestrado / Periodontia / Mestra em Clínica Odontológica

Osteomyelitis or bone infections remain very difficult to treat despite advances in treatment regimens and surgical technics. The bone microenvironment and compromised vasculature in addition to infected prosthesis and implants that were put in the bone during prior surgery impedes the antibiotic partition into the bone from systemic therapy in many cases. Treatment often includes surgical debridement of the infected bone and surrounding tissue, removal of implants, systemic antibiotic therapy accompanied with antibiotic containing bone void filler, in most cases polymethylmethacylate (PMMA) based bone cement. Unfortunately, PMMA has many associated problems, including non-biodegradability, inconsistent antibiotic release, and a surface susceptible to bacterial biofilm growth, ultimately necessitating removal and causing recurrent infections. Thus, recent studies have focused on designing novel bone void filling materials to deliver antibiotics and to support bone regeneration. There are two parts to designing a successful bone void filling device/material:(1) local release antibiotic for infection treatment and (2) development of a bone graft substitute to support bone regrowth. In this study, antibiotic releasing bone void filler (ABVF) putty formulations have been designed and tested. Different formulations were examined in this dissertation to describe the three components of the putty formulation - polymer, drug, and substrate. In the first formulation, different custom-made polymers were used to control drug release; Pro Osteon, a hydroxyapatite (HA) and calcium carbonate based bone graft substitute was used to provide support for bone growth. Finally, vancomycin was used as the antibiotic as it is clinically used to treat Staphylococcus aureus, the primary cause of osteomyelitis. In second formulation, commercially available and clinically used polymers, poly(lactic-co-glycolic acid) (PLGA), polycaprolactone (PCL) and, polyethylene glycol (PEG), were used to make the ABVF putty along with Pro Osteon and vancomycin. In the subsequent formulations, delivering combination antibiotics - vancomycin and rifampicin - to treat biofilm infections and, using bioglass (BG) as the substrate for faster bone regrowth were explored; PLGA, PCL and PEG constituted the polymer matrix. The ABVF putty formulations were customizable in terms of three primary components: polymers, bone graft substitutes, antibiotics. Ultimately, these were successful in curing infection and providing bone growth support.

No description available.