Peripheral vascular diseases such as peripheral artery disease (PAD) and critical limb ischemia (CLI) are growing at an ever-increasing rate in the Western world due to an aging population and the incidence of type II diabetes. A growing economic burden continues because these diseases are common indicators of future heart attack or stroke. Common therapies are generally limited to pharmacologic agents or endovascular therapies which have had mixed results still ending in necrosis or limb loss. Therapeutic angiogenic strategies have become welcome options for patients suffering from PAD due to the restoration of blood flow in the extremities. Capillary sprouting and a return to normoxic tissue states are also demonstrated by the use of angiogenic cytokines in conjunction with bone marrow cell populations. To this point, it has been determined that spatial and temporal controlled release of growth factors from vehicles provides a greater therapeutic and angiogenic effect than growth factors delivered intramuscularly, intravenously, or intraarterialy due to rapid metabolization of the cytokine, and non-targeted release. Furthermore, bone marrow cells have been implicated to enhance angiogenesis in numerous ischemic diseases due to their ability to secrete angiogenic cytokines and their numerous cell fractions present which are implicated to promote mature vessel formation. Use of angiogenic peptides, in conjunction with bone marrow cells, has been hypothesized in EPC mobilization from the periphery and marrow tissues to facilitate neovessel formation. For this purpose, controlled release of angiogenic peptides basic fibroblast growth factor (FGF-2) and granulocyte-colony stimulating factor (G-CSF) was performed using tunable ionic gelatin hydrogels or fibrin scaffolds with ionic albumin microspheres. The proliferation of endothelial cell culture was determined to have an enhanced effect based on altering concentrations of growth factors and method of release: co-delivery versus sequential. Scaffolds with these angiogenic peptides were implanted in young balb/c mice that underwent unilateral hindlimb ischemia by ligation and excision of the femoral artery. Endpoints for hindlimb reperfusion and angiogenesis were determined by Laser Doppler Perfusion Imaging and immunohistochemical staining for capillaries (CD-31) and smooth muscle cells (alpha-SMA). In addition to controlled release of angiogenic peptides, further studies combined the use of a fibrin co-delivery scaffold with FGF-2 and G-CSF with bone marrow stem cell transplantation to enhance vessel formation following CLI. Endpoints also included lipophilic vascular painting to evaluate the extent of angiogenesis and arteriogenesis in an ischemic hindlimb. Tissue engineering strategies utilizing bone marrow cells and angiogenic peptides demonstrate improved hindlimb blood flow compared to BM cells or cytokines alone, as well as enhanced angiogenesis based on immunohistochemical staining and vessel densities.

Spinal cord injuries result in permanent loss of motor function, leaving those affected with long term physical and financial burdens. Strategies for spinal cord injury repair must overcome unique challenges due to scar tissue that seals off the injury site, preventing regeneration. Tissue engineering can address these challenges with scaffolds that serve as cell- and drug-delivery tools, replacing damaged tissue while simultaneously addressing the inhibitory environment on a biochemical level. To advance this approach, the choice of cells, biomaterial matrix, and drug delivery system must be investigated and evaluated. This research seeks to evaluate (1) the behaviour of murine induced pluripotent stem cells in previously characterized 3D fibrin matrices; (2) the 3D fibrin matrix as a platform to support the differentiation of human induced pluripotent stem cells; and (3) the ability of an affinity-based drug delivery system to control the release of emerging spinal cord injury therapeutic, heat shock protein 70 from fibrin scaffolds. / Graduate / 0541 / amy.lynn.montgomery@gmail.com

Diss. (sammanfattning) Stockholm : Karol. inst., 2004. / Härtill 6 uppsatser.

Made available in DSpace on 2014-06-11T19:33:28Z (GMT). No. of bitstreams: 0 Previous issue date: 2003-06-18Bitstream added on 2014-06-13T19:04:26Z : No. of bitstreams: 1 theodoro_lh_dr_arafo.pdf: 4192650 bytes, checksum: 68b3ce4f45242d419b74abad0df09354 (MD5) / O objetivo deste estudo foi avaliar in vitro, a adesão de elementos sanguíneos sobre superfícies radiculares irradiadas com laser de Er:YAG e de Diodo, e a ação destes sobre as superfícies radiculares. Foram obtidas 120 amostras de dentes humanos extraídos por doença periodontal, que foram previamente raspados e aplainados com instrumentos manuais, sendo a seguir divididas aleatoriamente em 6 grupos com 20 amostras cada. O G1 (controle)- não recebeu nenhum tratamento; o G2- recebeu aplicação tópica de EDTA 24%; G3- foi irradiado com laser de Er:YAG com 7,6 J/cm2; o G4- irradiado com laser de Er:YAG com 12,9 J/cm 2; o G5 -irradiado com laser de Diodo com 90 J/cm2 e o G6 - foi irradiado com laser de Diodo com 108 J/cm2. Após a realização dos tratamentos, em 10 amostras de cada grupo foi depositado imediatamente a sua punção, tecido sanguíneo originado da vascularização periférica de humano, sendo que 10 amostras não receberam tal tratamento. Após processamento laboratorial as amostras foram analisadas através de microscopia eletrônica de varredura. As fotomicrografias obtidas foram avaliadas através de dois índices: adesão de elementos sanguineos e de morfologia da superfície radicular e estatisticamente analisadas (Kruskall Wallis e Mann-Whitney). Os resultados demonstraram que em relação a adesão de elementos sanguíneos não houve diferenças estatisticamente significante entre o grupo controle e os tratados com o laser de Er:YAG (p=0,7733 e 0,7831) ; O G5 mostrou-se menos efetivo que o controle na adesão de elementos sanguíneos ( p=0,0305 ) e o G2 foi o menos efetivo de todos na adesão. Com relação a morfologia da superfície radicular houve diferenças significantes entre o controle e os grupos do laser de Er:YAG (p= 0,0001) e entre o G5 do Diodo (0,0259); entre o EDTA e os grupos do laser de Er:YAG (p=0,0150 ) e entre o... . / The aim of this study was to evaluate in vitro the adhesion of blood components on root surfaces irradiated with Er:YAG and Diode lasers and these lasers effects on root surfaces. It was obtained 120 samples of human teeth extracted by periodontal disease. They were planed and scaled previously with manual instruments and randomly divided into 06 groups of 20 samples each. The groups were treated according to the following procedures: G1 (Control Group) -received no treatment; G2- had a topical application of EDTA 24%; G3- was irradiated with Er:YAG laser (7,6 J/cmø); G4 - was irradiated with Er:YAG laser (12,9 J/cmø); G5 - was irradiated with Diode laser (90 J/cmø) and G6 was irradiated with Diode laser (108 J/cmø). After these treatments were conducted, 10 samples of each group received a stippler, a blood tissue originated from peripheral vascularization, and the reminiscent samples did not receive such treatment. After laboratorial analysis the samples were analyzed through a scanning electronic microscopy. The photomicrographs obtained were evaluated according to adhesion of blood components and morphology of root surface; and statistically analyzed (Kruskall Wallis and Mann-Whitney). With relation to the adhesion of blood components, the results have shown that there were no significant differences between the Control Group and the groups treated with Er:YAG laser (p=0,7733 and 0,7831); G5 was less effective than the Control Group in adhesion of blood components (p=0,0305) and G2 was the least effective of all groups in this case. With relation to the morphology of root surface there were significant differences among the Control Group, the Er:YAG laser groups (p=0,0001) and the Diode laser G5 (p=0,0259); between the EDTA and the Er:YAG laser groups (p=0,0150) and also between the Diode laser G6 and the Er:YAG laser groups (p=0,0032)... (Complete abstract, click electronic address below).

Tissue engineering is an emerging interdisciplinary field developing new ways of treatment of patient's tissue defects using artificial substitutes. Skin tissue engineering is developing skin substitutes and wound dressings that would replace current treatment using autologous, allogeneic or xenogenic substitutes. There are high demands on materials which should serve as a scaffolds for dermal fibroblasts and keratinocytes. They must be non-cytotoxic and biodegradable with a rate proportional to formation of a new tissue. The materials should support adhesion and proliferation of the cells and even they could release growth factors and antimicrobial substance to enhance healing and new tissue formation. In this master thesis, the cell adhesion and proliferation were evaluated on sodium carboxymethyl cellulose (Hcel® NaT), poly-ε-caprolactone (PCL), poly-L-lactide-co-ε-caprolactone (PLA/PCL) and cellulose acetate (AC) nanofiber membranes. Primary human dermal fibroblasts and HaCaT cell line keratinocytes were selected for evaluation. The cell adhesion was observed by fluorescent microscopy, the proliferation was determined by metabolic assay (WST-1) and the material cytotoxicity was evaluated in xCELLigence® system. Materials did not show cytotoxic effects on the cells. However, the materials did...

Made available in DSpace on 2014-06-11T19:28:03Z (GMT). No. of bitstreams: 0 Previous issue date: 2006-03-10Bitstream added on 2014-06-13T19:57:17Z : No. of bitstreams: 1 leite_frm_me_arafo.pdf: 3458261 bytes, checksum: 24ab9b956cd575d2cc355dd7652fbe26 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Universidade Estadual Paulista (UNESP) / A raspagem gera smear layer que contém microrganismos e toxinas, podendo interferir no reparo periodontal. Por esse motivo, diferentes substâncias têm sido empregadas para remover esta camada e expor fibras colágenas da superfície dental. A adesão de elementos sangüíneos a superfícies radiculares desmineralizadas e a estabilização do colágeno pelas fibras colágenas são de extrema importância no sucesso da cirurgia periodontal. O objetivo deste estudo foi avaliar os diferentes padrões de adsorção e adesão de elementos sangüíneos a superfícies radiculares quimicamente condicionadas. Setenta e cinco dentes foram raspados, eqüitativamente divididos em 5 grupos: irrigação com água destilada (controle), aplicação de solução de ácido cítrico a 25%, solução de citrato de sódio a 30%, gel de EDTA a 24% e solução de cloridrato de tetraciclina a 50 mg/mL. Metade da superfície condicionada foi exposta a sangue fresco e preparada para microscopia eletrônica de varredura. As superfícies radiculares raspadas e condicionadas com EDTA e ácido cítrico apresentaram os melhores resultados em relação ao grupo controle. O ácido cítrico também se mostrou mais efetivo na remoção de smear layer e na adesão de elementos sangüíneos do que o cloridrato de tetraciclina e o citrato de sódio. Além disso, a relação entre exposição de fibras colágenas e adesão de elementos sangüíneos foi positiva. Dessa forma, o emprego de EDTA e ácido cítrico sobre a superfície radicular aumentam a adsorção e adesão de células sangüíneas e a estabilização da rede de fibrina. / Root scaling generates a smear layer which contains microorganisms and toxins that could interfere in periodontal healing. For this reason, different substances have been used to remove it and to expose collagen fibers from tooth surface. Blood elements adhesion to demineralized roots and clot stabilization by collagen fibers are extremely important for the success of periodontal surgery. The aim of this study was to evaluate the different patterns of blood elements adsorption and adhesion to root surfaces chemically conditioned. Seventy five teeth were planed and equitably divided into five groups: irrigation with distilled water (control), application of a 25% citric acid solution, 30% sodium citrate solution, 24% EDTA gel and 50 mg/mL tetracycline hydrochloride. Half of the conditioned surface was exposed to fresh blood and prepared for scanning electron microscopy. Planed root surfaces and conditioned with EDTA and citric acid showed better results when compared to control group. Citric acid was more effective on smear layer removal and blood elements adhesion to root surface than tetracycline and sodium citrate. Also, the relationship between collagen fibers exposure and blood elements adhesion was positive. This way, EDTA and citric acid employment on root surface improve blood element adsorption and adhesion to root surface and fibrin network stabilization.

Submitted by CLAUDIA VILALVA CASSARO null (claudia.v.cassaro@gmail.com) on 2018-03-21T13:51:17Z No. of bitstreams: 1 dissertacao claudia_cassaroFINAL.pdf: 3809022 bytes, checksum: a5c8891039ce6bc569b42ee66058c846 (MD5) / Rejected by Luciana Pizzani null (luciana@btu.unesp.br), reason: Solicitamos que realize uma nova submissão seguindo as orientações abaixo: Necessário fazer as seguintes correções no arquivo submetido: problema 1: Assinalar a agência de fomento (no seu caso CAPES). problema 2: No campo número do financiamento: colocar o número do processo. Assim que tiver efetuado as correções submeta o arquivo em PDF novamente. Agradecemos a compreensão. on 2018-03-22T12:35:28Z (GMT) / Submitted by CLAUDIA VILALVA CASSARO null (claudia.v.cassaro@gmail.com) on 2018-03-22T15:52:41Z No. of bitstreams: 1 dissertacao claudia_cassaroFINAL.pdf: 3809022 bytes, checksum: a5c8891039ce6bc569b42ee66058c846 (MD5) / Approved for entry into archive by ROSANGELA APARECIDA LOBO null (rosangelalobo@btu.unesp.br) on 2018-03-23T12:00:14Z (GMT) No. of bitstreams: 1 cassaro_cv_me_bot.pdf: 3809022 bytes, checksum: a5c8891039ce6bc569b42ee66058c846 (MD5) / Made available in DSpace on 2018-03-23T12:00:14Z (GMT). No. of bitstreams: 1 cassaro_cv_me_bot.pdf: 3809022 bytes, checksum: a5c8891039ce6bc569b42ee66058c846 (MD5) Previous issue date: 2018-02-23 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / O reparo do tecido ósseo é um desafio antigo da ciência mundial. A partir das décadas de 70-80, os biomateriais passaram a ser explorados e hoje desempenham importante papel na engenharia tecidual. Estes, quando aplicados na enxertia óssea, devem ser bioativos, biocompatíveis e reabsorvíveis. A integração entre os diversos materiais de origem sintética e os arcabouços biológicos possui amplo espectro de uso. Dentre os arcabouços destacam-se os compostos à base de fibrina por suas propriedades hemostáticas, estabilidade e morfologia favorável à proliferação celular. A presente revisão abordou a estrutura e constituição do tecido ósseo e suas limitações no processo de reparo, a relevância da engenharia tecidual aplicada à esta finalidade e os biomateriais utilizados para otimizar tal processo, com ênfase especial aos materiais compostos por fosfatos de cálcio, células tronco mesenquimais e sua aplicação junto ao arcabouço biológico selante de fibrina, com destaque para o selante heterólogo de fibrina, agora denominado biopolímero de fibrina, um produto único e em fase de testes em experimentação animal e estudos clínicos. Propõe-se, ao final desta, uma nova abordagem para a regeneração de defeitos ósseos de ratos baseada na associação entre o selante heterólogo de fibrina, o fosfato de cálcio bifásico e células tronco mesenquimais. / Bone tissue repair is an ancient challenge of world science. From the 70s and 80s, biomaterials began to be explored and nowadays play an important role in tissue engineering. These, when applied in bone grafting, have to be bioactive, biocompatible and resorbable. The integration between the various materials of synthetic origin and biological scaffolds has a wide spectrum of use. Among these scaffolds, fibrin-based compounds are distinguished by their hemostatic properties, stability and morphology favorable to cell proliferation. The present review addressed the structure and constitution of bone tissue and its limitations in the repair process, the relevance of tissue engineering approaches applied to this purpose and the biomaterials used to optimize this process, with special emphasis on materials composed of calcium phosphates, mesenchymal stem cells and its application with the biological sealant fibrin scaffold, highlighting the heterologous fibrin sealant, now named as fibrin biopolymer, a unique product testing in animal experimentation and clinical studies. At the end of this, a new approach is proposed for the regeneration of rat bone defects based on the association between heterologous fibrin sealant, biphasic calcium phosphate and mesenchymal stem cells.

Orientador: Rosemary Adriana Chiérici Marcantonio / Banca: Márcio Zafalon Casati / Banca: Estela Sasso Cerri / Resumo: Foram utilizados 60 dentes humanos nessa pesquisa. No primeiro experimento, 15 dentes extraídos por doença periodontal foram selecionados para avaliar a influência da irradiação com laser de Er,Cr: YSGG sobre a morfologia e adesão de células sanguíneas sobre as superfícies radiculares. As 60 amostras provenientes desses dentes foram divididos em 3 grupos de acordo com o tipo de tratamento aplicado: Grupo 1- RAR; Grupo 2- Irradiação com laser de Er,Cr: YSGG; Grupo 3- RAR e irradiação com o laser de Er,Cr: YSGG. 10 amostras de cada grupo foram avaliados quanto a adesão de elementos sanguíneos, e as outras 10 amostras foram avaliados quanto a morfologia da superfície radicular por MEV. Os testes de Kruskall-Wallis e Mann-Whitney foram utilizados para avaliar os resultados. Em relação à adesão de elementos sanguíneos, este estudo não demonstrou diferenças estatísticas entre os grupos (p=0.359), a análise morfológica demonstrou que as superfícies radiculares irradiadas com o laser de Er-Cr:YSGG foram mais rugosas que as do grupo controle (G2-G1: p=0.0003 e G3-G1: p=0.0003). No segundo experimento, 20 dentes foram utilizados para avaliar a influência do ângulo de irradiação do laser de Er,Cr:YSGG sobre a rugosidade e o desgaste das superfícies radiculares. Cada face proximal foi dividida em 3 áreas, sendo que a área superior foi tratada com raspagem e alisamento radicular, a área média não foi submetida a nenhum tipo de tratamento e a área inferior foi irradiada com o laser de Er,Cr:YSGG. Os dentes foram divididos em 4 grupos ,com 5 dentes cada, a depender da angulação da aplicação da irradiação do laser de Er,Cr:YSGG na área ( 30º, 45º, 60º, 90º). A rugosidade das áreas foram avaliadas através de um rugosímetro e posteriormente os dentes foram submetidos a processamento histológico... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: 60 human teeth were used in that research. In the first experiment, 15 extracted teeth for periodontal disease were selected to evaluated the effect of Er,Cr:YSGG irradiation on root surfaces for adhesion of blood components and morphology. 60 root surface specimens were obtained by selecting four from each tooth. Samples were divided into three groups of 20 each, according to treatments. Group 1 (G1) was treated by scaling and root planing (SRP), Group 2 (G2) was irradiated by Er,Cr:YSGG laser and Group 3 (G3) was treated by SRP and Er,Cr:YSGG laser irradiation. Blood was placed on each of 10 specimens from each of the three groups, to evaluate adhesion of blood components to the root surfaces. A morphological analysis was made of the root surfaces of the other 10 specimens from each group by scanning electron microscope (SEM). Statistical processing was done with the Kruskal-Wallis and Mann-Whitney tests. No statistical differences for adhesion of blood components to root surfaces were found between the groups (p = 0.359). However, morphological analysis disclosed that all root surfaces irradiated by Er,Cr:YSGG laser (100%) were rougher than surfaces that were not irradiated (G1-G2: p = 0.0003 and G1-G3: p = 0.0003). In the second experiment, 20 teeth were used to evaluated the effect of the working tip angulations on root roughness and substance removal using Er,Cr:YSGG radiation. The distal and mesial surfaces of each tooth was divided in 3 areas. The upper area was treated with scaling and root planing. The medium area was not submitted to any treatment and the lower area was irradiated with Er,Cr:YSGG laser. The teeh were divided in 4 groups, with 5 teeth each depending the working tip angulations using Er,Cr:YSGG at the lower area (30º, 45º, 60º, 90º).The roughness surfaces were evaluated by a profilometer, and... (Complete abstract, click electronic access below) / Mestre

INTRODUÇÃO: O tratamento padrão para lesões de nervo periférico que não podem ser suturados primariamente é a enxertia de nervo autólogo. Esse método, porém, carece de resultados satisfatórios e impõe algumas limitações técnicas e complicações. Várias opções já foram estudadas como alternativas ao enxerto de nervo, porém ainda não há conduto biológico ou sintético disponível para uso clínico que tenha a mesma capacidade regenerativa do enxerto de nervo autólogo. Os avanços em cultura celular e o maior entendimento dos mecanismos moleculares e celulares da regeneração nervosa levaram ao uso de células promotoras de regeneração associado aos condutos na tentativa de melhorar os resultados da reconstrução nervosa. Vários estudos demonstraram que o uso de célulastronco derivadas de tecido adiposo (ADSC) em condutos aloplásticos potencializa a regeneração neural. No entanto, nenhum estudo até hoje comparou a adição de ADSC indiferenciadas em conduto aloplástico ao tratamento padrão com autoenxerto. Esse estudo tem como objetivo avaliar a influência da adição de células-tronco mesenquimais derivadas de tecido adiposo em conduto de fibrina na regeneração de nervo periférico e comparar com enxertia de nervo autógeno em modelo experimental de ratos. MÉTODO: Em um modelo de lesão de nervo ciático (defeito de 10 mm) foram avaliados 30 ratos Wistar divididos em 3 grupos. O defeito de nervo foi reconstruído usando conduto de fibrina (Grupo Conduto, n=10), conduto de fibrina acrescido de ADSC (Grupo ADSC, n=10) e autoenxerto do nervo (Grupo Autoenxerto, n=10). A avaliação funcional dos ratos foi realizada com o teste de marcha (walking track analysis) com 4, 8 e 12 semanas e o índice de função ciática (IFC) foi determinado. Após 12 semanas, o peso do músculo tríceps sural foi avaliado. Segmentos dos nervos regenerados também foram coletados para análises histológicas como densidade axonal e diâmetro médio das fibras. RESULTADOS: O grupo Conduto mostrou recuperação funcional no teste da marcha após a reconstrução do nervo, porém com resultados inferiores aos outros dois grupos. O grupo ADSC mostrou recuperação intermediária e o grupo Autoenxerto obteve os melhores resultados (IFC com 12 semanas de -53,3±.3 vs -44,7±3 vs - 35,6±2, respectivamente, p < 0,001). A relação de peso do músculo tríceps sural no grupo Conduto foi de 41,1±3%, no grupo ADSC de 53,3±4% e no grupo Autoenxerto de 71,0 ± 4% (p < 0,001). Na avaliação histológica, o grupo Conduto mostrou densidade axonal de 39,8±3 axônios/10.995?m2 e diâmetro médio das fibras de 3,9 ± 0?m2, o grupo ADSC densidade axonal de 58,8 ± 3 axônios/10.995um2 e diâmetro médio das fibras de 4,9 ± 1um2 e o grupo Autoenxerto densidade axonal de 67,1±2 axônios/10.995?m2 e diâmetro médio das fibras de 8,9±1um2 (p < 0,001). CONCLUSÃO: A adição de células-tronco mesenquimais derivadas de tecido adiposo (ADSC) em conduto de fibrina na regeneração de nervo periférico, em modelo experimental de ratos, mostrou recuperação funcional e regeneração histológica estatisticamente mais significativa comparada à reconstrução somente com conduto de fibrina, porém ainda aquém dos resultados obtidos com enxertia de nervo autógeno / Introduction: The standard treatment for peripheral nerve injuries that cannot be primarily sutured is nerve autograft. But this method lacks satisfactory results and imposes some technical limitations and complications. Several options have been studied as alternatives to nerve autografting, but there is no biological or synthetic conduit available for clinical use that provides the same regenerative capacity of nerve autograft. Advances in cell culture and understanding of nerve regeneration mechanisms led to the use of regeneration-inducing cells in association with conduits, in an attempt to improve the reconstruction results. Several studies have shown that the use of adipose derived stem cells (ADSC) into conduits enhances neural regeneration. However, there is no study that compared the addition of undifferentiated ADSC in alloplastic conduit to standard treatment with autograft. This study evaluated the influence of the addition of adipose derived stem cell in fibrin conduit for peripheral nerve regeneration in comparison to the nerve autograft, in a rat model. Method: A sciatic nerve injury model (10-mm defect) was performed in 30 Wistar rats, which were divided into 3 groups. Nerve defect was reconstructed using fibrin conduit (Conduit group, n=10), fibrin conduit filled with ADSC (ADSC group, n = 10) and nerve autograft, (Autograft group, n=10). The walking behavior was measured by footprint analysis at 4, 8, and 12 weeks and sciatic function index (SFI) was determined. After 12 weeks, the triceps surae muscle weight was evaluated and histological analysis was performed to evaluate the regenerated nerve and measured axonal density and fibers diameter average. Results: The Conduit group showed less improvement in walking behavior compared to ADSC group and Autograft group (SFI at 12 weeks, - 53.3 ± .3 vs -44.7 ± 3 vs -35.6 ± 2 respectively, p< 0.001). The triceps surae muscle weight ratio of the fibrin conduit group was 41.1± 3%, ADSC group was 53.3 ± 4%, and Autograft group 71.0 ± 4% (p < 0.001). In histological evaluation, the Conduit group showed axonal density of 39.8±3 axons/10995um2 and fiber diameter average of 3.9±0 ?m2, the ADSC group had axonal density of 58.8 ± 3 axons/10995 um2 and fiber diameter average of 4.9±1?m2 and axon density of Autograft group was 67.1±2 axons/10995 um2 and fiber diameter average was 8.9±1?m2 (p < 0.001). Conclusion: The addition of adipose derived stem cells (ADSC) into fibrin conduit used for nerve reconstruction following peripheral nerve injury in the rat model, showed better functional recovery and better histological regeneration compared to reconstruction with fibrin conduit without ADSC. However, the functional recovery in the ADSC group was worse than that in nerve Autograft group and the nerve repair with the ADSC-fibrin conduit has less myelinated fibers when compared to the repair with nerve autograf

L’ingénierie tissulaire vise à régénérer des organes ou des tissus lésés. Ainsi, les gels de fibrine sont largement utilisés pour la reconstruction de différents tissus. Cependant, à concentration physiologique, ils ne peuvent pas être manipulés. Pour améliorer leurs propriétés mécaniques, ils peuvent être combinés dans une architecture de Réseaux Interpénétrés de Polymères (RIP) à un réseau de polymère synthétique (PVA ou POE). Ces RIPs peuvent être rendus biodégradables en copolymérisant d’albumine bovine de sérum modifiée par des groupements méthacrylate (BSAm) avec le partenaire synthétique.Selon leurs compositions, ces matériaux peuvent être complètement dégradés ou fragmentés après quelques jours en présence de thermolysine, une enzyme protéolytique. Ces cinétiques de dégradation de ces RIPs ont été étudiées en suivant le relargage des fragments protéiques hors du matériau, d’une part, et la diminution de leurs propriétés viscoélastiques, d’autre part. Leur biocompatibilité a été vérifiée : des fibroblastes cultivés à leur surface présentent une viabilité supérieure à 90% après 5 semaines de culture et leur prolifération est suivie de la synthèse de macromolécules de la Matrice Extracellulaire.Afin de leur ajouter une action bactéricide et d’augmenter encore leur résistance mécanique, des sels d’ammonium ont également été incorporés à certains de ces RIPs. Enfin, la synthèse de tels RIP a été mise au point à partir de plasma sanguin. Les éventuels phénomènes de rejet lors de leur intégration au sein du corps devraient ainsi être limités. De plus, le plasma sanguin contenant un grand nombre de facteurs de croissance et de molécules bioactives, la réparation tissulaire devrait ainsi être améliorée. / Tissue engineering aims to regenerate deficient tissues and organs. Fibrin gels are widely used for the reconstruction of various tissues. However, at physiological concentration, they can’t be handled. To improve their mechanical properties, they can be combined with a synthetic polymeric network (PVA or PEO) in an Interpenetrating Polymer Network (IPN) architecture. These IPN can be made biodegradable by crosslinking the Bovine Serum Albumin modified by methacrylate groups (BSAm) with the synthetic partner.Depending on the composition, these materials can be fully degraded or fragmented after several days of incubation with thermolysin, a proteolytic enzyme. The degradation kinetics of these hydrogels were studied by following the release of proteic fragments from the material and by the loss of viscoelastic properties. The biocompatibility was also verified: fibroblasts cultivated on the surface show a viability of over 90% after 5 weeks of culture and the proliferation is followed by the synthesis of Extracellular Matrix macromolecules.To add a bactericide property, and to increase their mechanical resistance, ammonium salts were incorporated in those IPN. Lastly, the synthesis of these IPN were adapted, starting from whole blood plasma. Rejection phenomena upon implantation should thus be hindered. Moreover, blood plasma contains a wide variety of growth factors and bioactive molecules, which should improve tissue regeneration.