No description available.

京都大学 / 新制・課程博士 / 博士(医学) / 甲第24481号 / 医博第4923号 / 新制||医||1063(附属図書館) / 京都大学大学院医学研究科医学専攻 / (主査)教授 高橋 淳, 教授 髙橋 良輔, 教授 井上 治久 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

No / In recent years, for neurodegenerative diseases therapy, research has focused on the stem cells therapy. Due to promising findings in stem cell therapy, there are various sources of stem cells for transplantation in human. The aim of this study was to evaluate sciatic nerve regeneration in the rat after nerve transaction followed by human endometrial stem cells (hEnSCs) treatment into poly (e-caprolactone)/collagen/nanobioglass (PCL/collagen/NBG) nanofibrous conduits. After treatment of animals, the performance in motor and sensory tests, showed significant improvement in rats treated with hEnSCs as an autograft. H&E images provided from cross-sectional and, longitudinal-sections of the harvested regenerative nerve as well as immunohistochemistry results indicated that regenerative nerve fibres had been formed and accompanied with new blood vessels in the conduit cell group. Due to the advantage of high surface area for cell attachment, it is reported that this electrospun nerve conduit could find more application in cell therapy for nerve regeneration in future, to further improve the functional regeneration outcome, especially for longer nerve defect restoration. In conclusion, our results suggest that the PCL/collagen/NBG nanofibrous conduit filled with hEnSCs is a suitable strategy to improve nerve regeneration after a nerve transaction in rat. / Iran National Science Foundation (INSF) grant number 95849510

Keratins are a family of structural proteins that can be extracted from a variety of sources including wool, nails, skin, hooves, and hair. Keratin can be processed into different constructs such as coatings, scaffolds, and hydrogels, and has shown favorable results when placed in in vitro and in vivo settings for different tissue regeneration applications. Over three decades, keratin extraction technology has been continuously modified, and these differences in extraction processes have distinct effects on the characteristics of the end product. In this work, we examine the effect of keratin aggregation during a widely-used purification step, dialysis ultra-filtration, on material characteristics of the final keratin product when fabricated into a hydrogel. Two distinct dialysis procedures were applied during the extraction of oxidized keratin (keratose): one promoting protein aggregation and the other mitigating it. Analyses of material properties such as mechanical and enzymatic stability were conducted in addition to observing the differences in solution behavior between products. Data revealed that protein aggregation during the extraction process has a profound effect on keratose hydrogel material properties. After determination of the effect of protein aggregation during extraction on keratose hydrogels, investigation of how a blended material comprised of said keratose and type I collagen was undertaken. It was hypothesized that a blend would result in mixing at the molecular level, resulting in improved properties compared to either pure material alone. A protocol was created to make stable keratose/type I collagen blends and material characterization techniques were applied to determine the inherent properties of samples with differing ratios. Crosslinking density, mechanical properties, enzymatic degradation properties, water uptake capacity, structural architecture, and thermal properties were all assessed. In addition, the ability of this material to maintain cell viability was conducted. Results showed that the addition of type I collagen has a significant effect on the properties of hydrogel blends with keratose compared to the pure keratose system. This was mostly evident with hydrogel mechanical stability and material architecture. Finally, the ability to use this hybrid material as a luminal filler for a nerve conduit during peripheral nerve regeneration was explored in an in vitro setting. The ability of this blend to promote Schwann cell viability was assessed in addition to determining the ability of these cells to attach and migrate through the material matrix. These experiments demonstrate proof-of-concept for the application of using keratose/type I collagen matrices as a luminal filler in peripheral nerve guidance conduits. / Doctor of Philosophy / Keratins are a family of structural proteins that can be extracted from wool, skin, nails, and hair, and that have been investigated in the field of tissue regeneration. Humans make several types of keratins, so it has a natural acceptance by the body and its inflammatory and immune systems. However, keratins can be hard to make and process into useful products. Many methods for producing keratin biomaterials have been developed over the past 30 years, but most of them are not ideal. This work sought to explore a production method that addresses a particular problem, that of protein aggregation during purification. In so doing, methods can be optimized to create more useful keratin biomaterials. Experiments comparing preparation methods that maximize and minimize protein aggregation were compared. Data showed that minimizing aggregation leads to better biomaterial characteristics, thus demonstrating the potential impact of targeting this processing step. However, even after optimization of purification, keratins still have limitations. Most notably their mechanical strength is not as great as some other materials. A typical approach to address this in other systems has been by blending. In the present work, we explored a blend made from keratin and type 1 collagen. A method was developed to effectively blend keratin and collagen and create stable mixtures that yielded protein-to-protein coordination. Such interactions typically yield beneficial material characteristics such as increased strength. Data showed that intimate mixing of the two proteins was achieved, and resulting characteristics were improved compared to either pure material. Finally, studies were conducted to assess the potential for keratin/collagen blends to be used to regenerate injured nerves. A common method is to enclose the ends of a cut nerve into a tube and let the nerve re-grow through the tube to its target muscle. An important characteristic is an ability for cells to populate the interior of the tube and help the nerve fibers grow. In the present study, we investigated the behavior of a particularly important cell, the Schwann cell, to attach, move and grow through a keratin/collagen biomaterial. Data showed good cell behavior, suggesting that the material could be used in a medical product for nerve repair.

Diss. (sammanfattning) Linköping : Univ., 2001. / Härtill 4 uppsatser.

Dans cette thèse, je me suis intéressé aux mécanismes qui permettent aux axones des neurones sensoriels de trouver leurs organes cibles à une grande distance. Dans le cas du système de la ligne latérale postérieure (LLP) du poisson-zèbre, des organes sensoriels sont déposés au cours de la migration d'un primordium. Des neurites sensoriels accompagnent le primordium au cours de cette migration et sont ainsi guidés vers leurs organes cibles. J'ai démontré que l'inactivation du signal «Glial cell line-Derived Neurotrophic Factor » (GDNF) rend les axones sensoriels incapables de suivre le primordium. GDNF est également utilisé comme signal de guidage lors de la régénération axonale après section du nerf et donc permet aux axones de retrouver leur cible. Ensuite j'ai démontré que le signal « Brain Derived Neurotrophic Factor » (BDNF) exerce un autre rôle dans le développement de la LLP puisqu'il est essentiel pour l'ancrage et la connexion des axones à leurs organes cibles. Dans une deuxième partie, nous avons montré que le développement de la LLP embryonnaire du Thon Rouge est fortement similaire à celui du Poisson-Zèbre, pourtant relativement basal. Cette similitude comprend le fait que les axones de la LLP suivent le primordium. / In this thesis, I address the question of how peripheral axons of sensory neurons find their distant target organs. In the case of the posterior lateral line (PLL) system of zebrafish, sensory organs are deposited by a migrating primordium and sensory neurites accompany this primordium during its migration. In this way, the neurites are guided to their prospective target organs. I show that the inactivation of «Glial cell line Derived Neurotrophic Factor » (GDNF) signaling leads to the inability of sensory axons to track the migrating primordium. GDNF signaling is also used as a guidance cue during axonal regeneration following nerve cut. I conclude that GDNF is a major determinant of directed neuritic growth and of target finding in this system, and propose that GDNF acts by promoting local neurite outgrowth. Further, I demonstrate that «Brain Derived Neurotrophic Factor » (BDNF) signaling exerts another role in PLL development as it is essent ial to anchor and properly connect axons to their targets organs.In another project, we could demonstrate that the development of the embryonic PLL of the atlantic blue-fin tuna shows striking similarities to that of the relatively basal zebrafish, including that PLL axons follow the migrating primordium.

This study examines the prospects of using the electrospinning process to fabricate tissue engineering scaffolds targeting a variety of regenerative applications, with a primary focus on the production of nerve guides for the treatment of long-defect nerve injuries in the peripheral nervous system. A basic overview of the conventional electrospinning process is provided, and the utility of this fabrication scheme in the production of collagen-based tissue engineering scaffolds is demonstrated. Next, a novel modification of the basic electrospinning process is presented. This process, called two pole air gap electrospinning, was developed to produce nerve guides that exhibit an anisotropic structure that mimics the extracellular matrix of native peripheral nerve tissue. This electrospinning process makes it possible to produce macroscopic nerve guides that are cylindrical in shape and composed of dense arrays of nano- to micron-scale diameter fibers. Unlike, conventional hollow core nerve guides, these electrospun constructs lack a central lumen, hence the designation 3D (for three-dimensional) nerve guide. The fibers are nearly exclusively arrayed in parallel with the long axis of the construct. This architectural feature provides thousands of individual channels, and aligned fibers that provide guidance cues that are designed to drive regenerating axons to grow in a highly directed fashion down the longitudinal axis of the guide. To supplement the structural cues provided by the fibrillar arrays of the electrospun 3D nerve guides, an alginate-based platform designed to deliver therapeutic reagents was developed and characterized. This platform makes it possible to fabricate gradients of therapeutic reagents within the fibrillar arrays of an electrospun nerve guide. Functional and structural analyses of these constructs supplemented with or without a gradient of NGF, in a long-defect nerve injury in the rodent sciatic nerve indicate that the 3D design is superior to the gold standard treatment, the autologous nerve graft. Animals treated with the 3D grafts recovered motor and sensory function faster and exhibited far higher nerve-to-nerve and nerve-to-muscle signal amplitudes in electrophysiological studies than animals treated with autologous grafts or conventional hollow core cylindrical grafts.

A tubulização tem sido considerada uma alternativa eficaz no reparo de lesões de nervos por permitir a aplicação local de fatores neurotróficos e facilitar estudos experimentais. A compreensão do mecanismo de ação no processo regenerativo é intrinsecamente dependente da identificação dos componentes moleculares ao longo dos processos degenerativo e regenerativo. A presente tese visa o estudo sequencial da identificação do perfil molecular da resposta imune no processo de regeneração de nervo entre duas técnicas microcirúrgicas: a tubulização látero-terminal (TLT) e a tubulização término-terminal (TTT). Ratas Wistar adultas foram submetidas à transecção bilateral do nervo fibular comum. Na técnica TLT: o nervo fibular foi seccionado e retirado 5mm de sua extensão, o coto proximal foi suturado na musculatura e um tubo de silicone vazio (6mm) foi interposto entre a lateral do nervo tibial (doador) e o coto distal do fibular. Na técnica TTT: o nervo fibular foi seccionado e retirado 5mm de sua extensão, e ambos os cotos foram suturados às extremidades do tubo de silicone vazio. Foram realizados os grupos experimentais: TLT convencional (TLT padrão), TLT com tubo fechado suturado a porção proximal (TLT F-P), TLT com tubo fechado suturado na porção distal (TLT FD), TTT convencional (TTT padrão), TTT com tubo fechado suturado ao coto proximal (TTT F-P), TTT com tubo fechado suturado ao coto distal (TTT F-D). O controle normal (grupo GN) foi o segmento de nervo retirado para a interposição do tubo. As análises foram realizadas em três tempos pós-cirurgia: 24 horas, 96 horas e 8 dias. As amostras foram processadas em teste comercial para identificação de anticorpos com intuito de determinar a intensidade de expressão de citocinas, quimiocinas e fatores de crescimento presentes no cone de crescimento dos diferentes grupos. Em 24h a expressão de citocinas foi similar entre os grupos, entretanto, o TLT apresentou maior número de quimiocinas expressas em relação ao TTT. O tempo de 96h foi o período com maior número de elementos que apresentaram aumento da expressão em ambos os grupos TLT e TTT, sendo o grupo TLT a apresentar número maior de quimiocinas e fatores de crescimento expressos. Em 8 dias, houve redução dos níveis de expressão de todos os elementos estudados em ambos os grupos, com exceção do CNTF que apresentou aumento de sua expressão no grupo TTT. Os grupos fechados proximais (TLT F-P e TTT F-P) apresentaram perfis de expressão distintos no tempo de 24h, não houve mudanças dos níveis de expressão de nenhum dos elementos do grupo TLT, sendo que, inversamente, o grupo TTT apresentou aumento da expressão de grande parte das proteínas estudadas. Em ambos os grupos TLT F-P e TTT F-P, os elementos investigados retornaram aos níveis basais de expressão em 96h e 8 dias. Os grupos fechados distais (TLT F-D e TTT F-D) apresentaram expressão mínima ou nenhuma dos elementos estudados. O presente estudo demonstrou que há diferenças quanto ao perfil molecular da resposta imune induzida por lesão de nervo entre as técnicas TLT e TTT. / Nerve tubulization has been considered an effective alternative to repair nerve damages due to allow the local application of growth factors and to facilitate experimental studies. The action mechanisms comprehension in nerve regeneration is intrinsically dependente of the molecular components identification over the degenerative and regenerative processes. The aim of this study was the sequencial analysis of the molecular identification of the imune response in nerve regeneration process between two microsurgical repair techniques: the end-to-side tubulization (EST) and the end-to-end tubulization (ETE). Bilateral transection of the common fibular nerve was done in adults female Wistar rats. EST model: the fibular nerve was sectioned and 5mm from his extension was removed, the proximal stump was sutured in the musculature and an empty silicone tube (6mm) was interposed between the lateral portion of the intact tibial donor nerve and the fibular nerve distal stump. ETE model: the fibular nerve was sectioned and 5mm from his extension was removed, both stumps were sutured to the silicone tube extremities. The experimentals groups done: EST conventional (EST standard), EST with closed tube sutured on the proximal portion (EST C-P), EST with closed tube sutured on the distal portion (EST C-D), ETE conventional (ETE standard), ETE with closed tube sutured on the proximal stump (ETE C-P), ETE with closed tube sutured on the distal stump (ETE C-D). The normal control (NG group) was the nerve extension removed to the tube interposition. The analyzes were performed in three times: 24 hours, 96 hours and 8 days. The samples were precessed on an antibody arrays to determine the expression intensity of cytokines, chemokines ans growth factors in the growth cone of the diferente groups. On 24h the cytokine expression was similar between groups, however, the ETE group presented a higher number of the expressed chemokines than ETS. The time 96h was the period with higher number of elements that presented increased expression in both groups EST and ETE, being the group ETS to presents higher number of the chemokines and growth factors expressed. In 8 days there was expression levels reduced of the all studied elements in both groups, with the exception of CNTF that presented increased expression on ETE group. The proximals closed groups (ETS C-P and ETE CP) presented different expression profiles on the time of 24h, there were no changes in the expression levels of any of the elements of the ETS group, whereas, conversely, the ETE group showed increased expression of a large part of the proteins studied. In both groups ETS C-P and ETE C-P, the investigated elements returned to baseline levels of expression at the 96h and 8 days dates. The distal closed groups (ETS C-D and ETE CD) presented minimal expression or no expression of the studied elements. This study demonstrates that there are differences referring to the molecular profile of the immune response induced by nerve damage between the ETS and ETE models.

A tubulização tem sido considerada uma alternativa eficaz no reparo de lesões de nervos por permitir a aplicação local de fatores neurotróficos e facilitar estudos experimentais. A compreensão do mecanismo de ação no processo regenerativo é intrinsecamente dependente da identificação dos componentes moleculares ao longo dos processos degenerativo e regenerativo. A presente tese visa o estudo sequencial da identificação do perfil molecular da resposta imune no processo de regeneração de nervo entre duas técnicas microcirúrgicas: a tubulização látero-terminal (TLT) e a tubulização término-terminal (TTT). Ratas Wistar adultas foram submetidas à transecção bilateral do nervo fibular comum. Na técnica TLT: o nervo fibular foi seccionado e retirado 5mm de sua extensão, o coto proximal foi suturado na musculatura e um tubo de silicone vazio (6mm) foi interposto entre a lateral do nervo tibial (doador) e o coto distal do fibular. Na técnica TTT: o nervo fibular foi seccionado e retirado 5mm de sua extensão, e ambos os cotos foram suturados às extremidades do tubo de silicone vazio. Foram realizados os grupos experimentais: TLT convencional (TLT padrão), TLT com tubo fechado suturado a porção proximal (TLT F-P), TLT com tubo fechado suturado na porção distal (TLT FD), TTT convencional (TTT padrão), TTT com tubo fechado suturado ao coto proximal (TTT F-P), TTT com tubo fechado suturado ao coto distal (TTT F-D). O controle normal (grupo GN) foi o segmento de nervo retirado para a interposição do tubo. As análises foram realizadas em três tempos pós-cirurgia: 24 horas, 96 horas e 8 dias. As amostras foram processadas em teste comercial para identificação de anticorpos com intuito de determinar a intensidade de expressão de citocinas, quimiocinas e fatores de crescimento presentes no cone de crescimento dos diferentes grupos. Em 24h a expressão de citocinas foi similar entre os grupos, entretanto, o TLT apresentou maior número de quimiocinas expressas em relação ao TTT. O tempo de 96h foi o período com maior número de elementos que apresentaram aumento da expressão em ambos os grupos TLT e TTT, sendo o grupo TLT a apresentar número maior de quimiocinas e fatores de crescimento expressos. Em 8 dias, houve redução dos níveis de expressão de todos os elementos estudados em ambos os grupos, com exceção do CNTF que apresentou aumento de sua expressão no grupo TTT. Os grupos fechados proximais (TLT F-P e TTT F-P) apresentaram perfis de expressão distintos no tempo de 24h, não houve mudanças dos níveis de expressão de nenhum dos elementos do grupo TLT, sendo que, inversamente, o grupo TTT apresentou aumento da expressão de grande parte das proteínas estudadas. Em ambos os grupos TLT F-P e TTT F-P, os elementos investigados retornaram aos níveis basais de expressão em 96h e 8 dias. Os grupos fechados distais (TLT F-D e TTT F-D) apresentaram expressão mínima ou nenhuma dos elementos estudados. O presente estudo demonstrou que há diferenças quanto ao perfil molecular da resposta imune induzida por lesão de nervo entre as técnicas TLT e TTT. / Nerve tubulization has been considered an effective alternative to repair nerve damages due to allow the local application of growth factors and to facilitate experimental studies. The action mechanisms comprehension in nerve regeneration is intrinsically dependente of the molecular components identification over the degenerative and regenerative processes. The aim of this study was the sequencial analysis of the molecular identification of the imune response in nerve regeneration process between two microsurgical repair techniques: the end-to-side tubulization (EST) and the end-to-end tubulization (ETE). Bilateral transection of the common fibular nerve was done in adults female Wistar rats. EST model: the fibular nerve was sectioned and 5mm from his extension was removed, the proximal stump was sutured in the musculature and an empty silicone tube (6mm) was interposed between the lateral portion of the intact tibial donor nerve and the fibular nerve distal stump. ETE model: the fibular nerve was sectioned and 5mm from his extension was removed, both stumps were sutured to the silicone tube extremities. The experimentals groups done: EST conventional (EST standard), EST with closed tube sutured on the proximal portion (EST C-P), EST with closed tube sutured on the distal portion (EST C-D), ETE conventional (ETE standard), ETE with closed tube sutured on the proximal stump (ETE C-P), ETE with closed tube sutured on the distal stump (ETE C-D). The normal control (NG group) was the nerve extension removed to the tube interposition. The analyzes were performed in three times: 24 hours, 96 hours and 8 days. The samples were precessed on an antibody arrays to determine the expression intensity of cytokines, chemokines ans growth factors in the growth cone of the diferente groups. On 24h the cytokine expression was similar between groups, however, the ETE group presented a higher number of the expressed chemokines than ETS. The time 96h was the period with higher number of elements that presented increased expression in both groups EST and ETE, being the group ETS to presents higher number of the chemokines and growth factors expressed. In 8 days there was expression levels reduced of the all studied elements in both groups, with the exception of CNTF that presented increased expression on ETE group. The proximals closed groups (ETS C-P and ETE CP) presented different expression profiles on the time of 24h, there were no changes in the expression levels of any of the elements of the ETS group, whereas, conversely, the ETE group showed increased expression of a large part of the proteins studied. In both groups ETS C-P and ETE C-P, the investigated elements returned to baseline levels of expression at the 96h and 8 days dates. The distal closed groups (ETS C-D and ETE CD) presented minimal expression or no expression of the studied elements. This study demonstrates that there are differences referring to the molecular profile of the immune response induced by nerve damage between the ETS and ETE models.

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