Use of aerobic treadmill exercises on nerve regeneration after sciatic nerve injury in spontaneously hypertensive rats

Purpose: Various postoperative protocols have been proposed to improve outcomes and accelerate nerve regeneration. Recently, the use of physical exercise in a post-surgical neurorraphy procedure has shown good results when started early. We experimentally investigated the hypothesis that post-operative exercise speeds up results and improves clinical and morphologic parameters. Methods: Isogenic rats were randomly divided into four groups: 1 SHAM; 2 SHAM submitted to the exercise protocol (EP); 3 Grafting of the sciatic nerve; and 4 Grafting of the sciatic nerve associated with the EP. The EP was based on aerobic activities with a treadmill, with a progressive increase in time and intensity during 6 weeks. The results were evaluated by the sciatic functional index (SFI), morphometric and morphologic analysis of nerve distal to the lesion, and the number of spinal cord motor neurons, positive to the marker Fluoro-Gold (FG), captured retrogradely through neurorraphy. Results: Functional analysis (SFI) did not show a statistical difference between the group grafted with (­50.94) and without exercise (-65.79) after 90 days. The motoneurons count (Spinal cord histology) also showed no diference between these groups (834.5 × 833 respectively). Although functionally there is no difference between these groups, morphometric study showed a greater density (53.62) and larger fibers (7.762) in GRAFT group. When comparing both operated groups with both SHAM groups, all values were much lower. Conclusions: The experimental model that this aerobic treadmill exercises protocol did not modify nerve regeneration after sciatic nerve injury and repair with nerve graft.

Stem cells in end-to-side neurorrhaphy: experimental study in rats

Purpose: To evaluate the influence of mesenchymal stem cells from adipose tissue in the end-to-side neurorrhaphy, focusing in the nerve regeneration and the muscle reinnervation in acute trauma. Methods: 140 animals were randomly divided in seven groups: control, denervated, end-to-side neurorrhaphy between distal stump of common peroneal nerve and tibial nerve (ESN), ESN wrapped in fascia, ESN wrapped in fascia and platelet gel, ESN wrapped in platelet gel, ESN wrapped in fascia and platelet gel within stem cells (without culture) removed from the adipose tissue. Mass measurements of the animal and of cranial tibial muscles, electromyography, walking track analysis tests and histological examinations of the nerves and muscles after 180 days was performed. Results: In the groups where the ESN was performed, the results were always better when compared to the denervated group, showing reinnervation in all ESN groups. The most sensitive methods were walking track and histological analysis. Only the group with stem cells showed values similar to the control group, as well as the functional indices of peroneal nerve and the number of nerve fibers in the peroneal nerve. Conclusions: Stem cells were effective in ESN according with the functional index of the peroneal nerve, evaluated by walking track analysis and the number of nerve fibers in the peroneal nerve.(AU)

Combination of heterologous fibrin sealant and bioengineered human embryonic stem cells to improve regeneration following autogenous sciatic nerve grafting repair

Background Peripheral nerve injury is a worldwide clinical problem, and the preferred surgical method for treating it is the end-to-end neurorrhaphy. When it is not possible due to a large nerve gap, autologous nerve grafting is used. However, these surgical techniques result in nerve regeneration at highly variable degrees. It is thus very important to seek complementary techniques to improve motor and sensory recovery. One promising approach could be cell therapy. Transplantation therapy with human embryonic stem cells (hESCs) is appealing because these cells are pluripotent and can differentiate into specialized cell types and have self-renewal ability. Therefore, the main objective of this study was to find conditions under which functional recovery is improved after sciatic nerve neurorrhaphy. We assumed that hESC, either alone or in combination with heterologous fibrin sealant scaffold, could be used to support regeneration in a mouse model of sciatic nerve injury and repair via autografting with end-to-end neurorrhaphy. Methods Five millimeters of the sciatic nerve of C57BL/6 J mice were transected off and rotated 180 degrees to simulate an injury, and then stumps were sutured. Next, we applied heterologous fibrin sealant and/or human embryonic stem cells genetically altered to overexpress fibroblast growth factor 2 (FGF2) at the site of the injury. The study was designed to include six experimental groups comprising neurorrhaphy (N), neurorrhaphy + heterologous fibrin sealant (N + F), neurorrhaphy + heterologous fibrin sealant + doxycycline (N + F + D), neurorrhaphy + heterologous fibrin sealant + wild-type hESC (N + F + W), neurorrhaphy + heterologous fibrin sealant + hESC off (N + F +T), and neurorrhaphy + heterologous fibrin sealant + hESC on via doxycycline (N + F + D + T). We evaluated the recovery rate using Catwalk and von Frey functional recovery tests, as well as immunohistochemistry analysis. Results The experiments indicated...

Combination of heterologous fibrin sealant and bioengineered human embryonic stem cells to improve regeneration following autogenous sciatic nerve grafting repair

Background Peripheral nerve injury is a worldwide clinical problem, and the preferred surgical method for treating it is the end-to-end neurorrhaphy. When it is not possible due to a large nerve gap, autologous nerve grafting is used. However, these surgical techniques result in nerve regeneration at highly variable degrees. It is thus very important to seek complementary techniques to improve motor and sensory recovery. One promising approach could be cell therapy. Transplantation therapy with human embryonic stem cells (hESCs) is appealing because these cells are pluripotent and can differentiate into specialized cell types and have self-renewal ability. Therefore, the main objective of this study was to find conditions under which functional recovery is improved after sciatic nerve neurorrhaphy. We assumed that hESC, either alone or in combination with heterologous fibrin sealant scaffold, could be used to support regeneration in a mouse model of sciatic nerve injury and repair via autografting with end-to-end neurorrhaphy. Methods Five millimeters of the sciatic nerve of C57BL/6 J mice were transected off and rotated 180 degrees to simulate an injury, and then stumps were sutured. Next, we applied heterologous fibrin sealant and/or human embryonic stem cells genetically altered to overexpress fibroblast growth factor 2 (FGF2) at the site of the injury. The study was designed to include six experimental groups comprising neurorrhaphy (N), neurorrhaphy + heterologous fibrin sealant (N + F), neurorrhaphy + heterologous fibrin sealant + doxycycline (N + F + D), neurorrhaphy + heterologous fibrin sealant + wild-type hESC (N + F + W), neurorrhaphy + heterologous fibrin sealant + hESC off (N + F +T), and neurorrhaphy + heterologous fibrin sealant + hESC on via doxycycline (N + F + D + T). We evaluated the recovery rate using Catwalk and von Frey functional recovery tests, as well as immunohistochemistry analysis. Results The experiments indicated...(AU)

Glial scar-modulation as therapeutic tool in spinal cord injury in animal models

Purpose: Spinal Cord injury represents, in veterinary medicine, most of the neurological attendances and may result in permanent disability, death or euthanasia. Due to inflammation resulting from trauma, it originates the glial scar, which is a cell interaction complex system. Its function is to preserve the healthy circuits, however, it creates a physical and molecular barrier that prevents cell migration and restricts the neuroregeneration ability. Methods: This review aims to present innovations in the scene of treatment of spinal cord injury, approaching cell therapy, administration of enzyme, anti-inflammatory, and other active principles capable of modulating the inflammatory response, resulting in glial scar reduction and subsequent functional improvement of animals. Results: Some innovative therapies as cell therapy, administration of enzymes, immunosuppressant or other drugs cause the modulation of inflammatory response proved to be a promising tool for the reduction of gliosis. Conclusion: Those tools promise to reduce gliosis and promote locomotor recovery in animals with spinal cord injury.(AU)

Effect of ozone and methylprednisolone treatment following crush type sciatic nerve injury

PURPOSE:To assess and compare the histopathological effects of ozone therapy and/or methylprednisolone (MPS) treatment on regeneration after crush type sciatic nerve injury.METHODS:Forty Sprague-Dawley male rats were randomly allocated into four groups. Four groups received the following regimens intraperitoneally every day for 14 days after formation of crush type injury on sciatic nerve: Group I: ozone (20mcg/ml); Group II: methylprednisolone (2mg/kg); Group III: ozone (20 mcg/ml) and methylprednisolone (2mg/kg); Group IV: isotonic saline (0.9%). The histomorphological evaluation was made after biopsies were obtained from the sites of injury.RESULTS:Significant differences were noted between groups in terms of degeneration (p=0.019), nerve sheath cell atrophy (p=0.012), intraneural inflammatory cellular infiltration (p=0.002), perineural granulation tissue formation (p=0.019), perineural vascular proliferation (p=0.004), perineural inflammatory cellular infiltration (p<0.001) and inflammation in peripheral tissue (p=0.006). Degeneration was remarkably low in Group III, while no change in nerve sheath cell was noted in Group II.CONCLUSION:The combined use of methylprednisolone and ozone treatment can have beneficial effects for regeneration after crush type nerve injury.(AU)

A Catuama e o bilobalide na regeneração nervosa periférica de ratos submetidos à secção do nervo isquiático / Catuama and bilobalide on peripheral nerve regeneration in rats following sciatic nerve section

A Catuama® é a associação de quatro extratos hidroalcoolicos obtidos de plantas brasileiras (Paullinia cupana, Trichilia catigua, Ptychopetalum olacoides e Zingiber officinale) com conhecida ação neuroprotetora, anti-inflamatória, antioxidante e antidepressiva. O bilobalide é um componente extraído das folhas do Ginkgo biloba, que tem comprovada ação neuroprotetora nos sistemas nervosos central e periférico. O presente estudo avaliou os efeitos da Catuama® e do bilobalide na regeneração nervosa periférica de ratos submetidos à secção do nervo isquiático. Foram utilizados 40 ratos com implante de tubo de silicone preenchido por colágeno líquido, deixando-se um intervalo entre os segmentos nervosos de 10mm. Os animais foram divididos em 4 grupos: o grupo controle (A); os grupos que receberam a Catuama® administrada por via oral nos primeiros 28 dias de pós-operatório, nas doses de 100 (B) e 400mg.kg-1 (C); e o grupo que recebeu o bilobalide na dose de 200µM (D), este, adicionado ao colágeno líquido utilizado no implante de silicone. Os animais foram avaliados na primeira, quinta e décima semanas de pós-operatório pelo teste de marcha. Na décima semana, foi realizada avaliação eletrofisiológica e análises quantitativa e qualitativa dos cortes histológicos de amostras do nervo isquiático e do músculo gastrocnêmio. Em todas as análises utilizadas observou-se excelente regeneração dos nervos, no entanto, não foi encontrada diferença significativa (P>0,05) entre os grupos experimentais e controle.(AU)

The Catuama® is composed of four Brazilian plants extracts (Paullinia cupana, Trichilia catigua, Ptychopetalum olacoides e Zingiber officinale). The Catuama® is known as having neuroprotector, anti-inflammatory, antioxidant and antidepressant effects. Bilobalide, extracted from leaves of Ginkgo biloba, is known by its neuroprotective effect in the central and peripheral nervous systems. The present study evaluates the effect of Catuama® and bilobalide on peripheral nerve regeneration in rats following a sciatic nerve section. Sciatic nerve of forty adult rats was transected with a 10-mm gap and the proximal and distal nerve stumps were fixed in a silicone tube filled with liquid collagen. The animals were divided into four groups: the control group (A), two groups treated with Catuama® by gavage along 28 days after the surgery in different doses of 100 (B) and 400mg.kg-1 (C) and the group using 200µM bilobalide (D) associated with the liquid collagen in the silicone tube. Evaluations were done by a walk test on the first, fifth and tenth week after the surgery. Electrophysiological stimulation and quantitative and qualitative histological analyses of the sciatic nerve and gastrocnemius muscle were also performed on the tenth week after the surgery. All groups showed good regeneration but no statistical difference was found between treatments and control groups (P>0.05).(AU)

Behavioural and cognitive effects of simvastatin dose used in stimulation of bone regeneration in rats

To analyze the effects of simvastatin (SVT) in the locomotion, anxiety and memory of rats, as a reflection of the administration of a minimum dose capable of stimulating bone regeneration in defects in the calvariae. METHODS: Surgical procedures were performed in 15 female Wistar rats, 2-month old, to insert the grafting material regenerator (Bone-ceramic(r)) and/or SVT, followed by behavioural and cognitive assessments in the 7th, 30th and 60th days post surgery. RESULTS: The SVT locally applied with the goal of bone regeneration in defects created in rat calvariae does not interfere with locomotion, anxiety levels and/or memories of rats, except for the first week following surgery, when an anxiolytic effect was observed, as a result of a possible central action. CONCLUSION: Failure to provoke any response within 30 and 60 days post surgical procedures suggests that SVT may constitute a good choice in stimulating bone regeneration without affecting the long term neural functions.(AU)

Perfil proteômico do líquido cefalorraquidiano após transplantes intratecal de células estromais mesenquimais multipotentes em equinos

Estudos com células estromais mesenquimais multipotentes (MSCs) estão em crescente progresso devido às suas propriedades imunomoduladoras, antiinflamatórias, antiapoptóticas e de regeneração tecidual, tornando essa modalidade de terapia celular promissora no tratamento de diversas doenças. Devido à limitada capacidade regenerativa do sistema nervoso central (CNS), causando sequelas funcionais, as MSCs estão sendo investigadas como uma alternativa terapêutica para condições neurológicas inflamatórias, vasculares, traumáticas e degenerativas em diversas espécies animais. A Mieloencefalite protozoária equina (EPM) causada por ambos os protozoários do filo Apicomplexa, Sarcocystis neurona e Neospora hughesi, permanece como uma importante doença neurológica dos equinos nas Américas, embora a maioria dos casos seja devida à infecção por S. neurona. A aplicação da proteômica com sua gama de ferramentas na clínica de equinos pode contribuir significativamente para o entendimento de processos patológicos e facilitar a descoberta de novos alvos terapêuticos ou marcadores diagnósticos. Neste contexto, os objetivos deste estudo foram avaliar o perfil proteômico do líquido cefalorraquidiano (CSF) antes e após múltiplos transplantes intratecal de MSCs em equinos hígidos e o perfil proteômico do CSF de equinos cronicamente afetados pela EPM. Doze cavalos adultos clinicamente saudáveis foram divididos aleatoriamente em três grupos experimentais: grupo DPBS (DPBS ou control; n = 4) onde a solução salina tamponada com fosfato de Dulbecco's (DPBS) foi administrada pela via intratecal; grupo AD-G (AD-G; n = 4), no qual foram realizados transplantes intratecal com MSCs alogênicas de tecido adiposo; e grupo BM-G (BM-G; n = 4), no qual foram realizados transplantes intratecal com MSCs alogênicas de medula óssea. Todos os grupos experimentais receberam três tratamentos seriados (DPBS ou MSCs) com intervalo de 30 dias entre eles. As amostras de CSF foram coletadas dos grupos experimentais imediatamente antes do primeiro tratamento (denominado momento M0 ou Before) e 30 dias após o terceiro tratamento (denominado momento M90 ou After). Ao mesmo tempo, os CSF de 16 equinos clinicamente saudáveis e de 9 equinos cronicamente afetados por EPM foram coletados da subaracnóide, denominados grupo Control (Control; n = 16) e grupo EPM (EPM; n = 9) respectivamente. Cada uma das amostras de CSF do grupo Control e do grupo EPM foram individualmente ressuspendidas e agrupadas em pool, totalizando dois pool representativos de cada um dos grupos experimentais e denominados igualmente (Control e EPM) para avaliar o perfil proteômico dos mesmos. De modo similar, os três grupos que receberam transplantes e coletas de CSF nos dois distintos momentos, foram agrupados em seis pool representativos para cada grupo e momento. Considerando a plataforma proteômica utilizada, os três grupos foram pareadamente comparados: DPBS After vs. Before, AD-G After vs. Before e BM-G After vs. Before, sendo encontradas 208, 211 e 131 proteínas em cada comparação, respectivamente. Do mesmo modo, na comparação pareada do CSF dos grupos EPM e Control, EPM vs. Control, 201 proteínas foram identificadas e 33 proteínas observadas exclusivamente no CSF do grupo EPM. Ademais, também foi observada na comparação pareada dos grupos DPBS, BM-G e EPM, a presença dos três tipos de enolases interagindo entre si (ENO 1, ENO 2 e ENO 3), sendo em DPBS e BM-G como exclusivas do momento After e no grupo EPM como exclusivas dele. Neste contexto, este estudo ao avaliar os perfis proteômico do CSF antes e após múltiplos transplantes intratecal de MSCs em equinos hígidos e do CSF de equinos cronicamente afetados pela EPM permitiu identificar as enolases como potenciais marcadores de lesão neural e/ou de EPM. Novos estudos devem ser realizados para confirmar estes achados e avançar no conhecimento dos efeitos da terapia celular com MSCs pela via intratecal, para o tratamento de lesões neurológicas em equinos.

Multipotent mesenchymal stromal cell (MSCs) studies are under increasing progress because of their immunomodulatory, anti-inflammatory, antiapoptotic and tissue regeneration properties, making this modality of cell therapy promising in the treatment of various diseases. Due to the limited regenerative capacity of the central nervous system (CNS), causing functional sequelae, MSCs are being investigated as a therapeutic alternative for inflammatory, vascular, traumatic and degenerative neurological conditions in various animal species. Equine protozoal myeloencephalitis (EPM) caused by both protozoa of the Apicomplexa phylum, Sarcocystis neurona and Neospora hughesi, remains an important neurological disease in horses in the Americas, although most cases are due to S. neurona infection. The application of proteomics with its range of tools in the equine clinic can contribute significantly to the understanding of pathological processes and facilitate the discovery of new therapeutic targets or diagnostic markers. In this context, the objectives of this study were to evaluate the proteomic profiling of cerebrospinal fluid (CSF) before and after multiple intrathecal transplantations of MSCs in healthy horses and the CSF proteomic profiling of horses chronically affected by EPM. Twelve clinically healthy adult horses were randomly divided into three experimental groups: DPBS (DPBS or control; n = 4), in which intrathecal "transplants" with Dulbecco's phosphate buffered saline (DPBS) were performed; group AD-G (AD-Gs; n = 4), in which intrathecal transplants were performed with allogeneic adipose tissue MSCs; and BM-G group (BM-G; n = 4), in which intrathecal transplants were performed with allogeneic bone marrow MSCs. All experimental groups received three serial treatments (DPBS or MSCs) with a 30-day interval between them. CSF samples were collected from the experimental groups immediately before the first treatment (called the M0 or Before) and 30 days after the third treatment (called the M90 or After). At the same time, CSF of 16 healthy horses and 9 horses chronically affected by EPM were collected from the subarachnoid, Control group (Control; n = 16) and EPM group (EPM; n = 9) respectively. Each of the CSF samples from the Control group and from the EPM group were individually resuspended and pooled, totalizing two representative pools of each one the experimental groups and also named (Control and EPM) to evaluate their proteomic profilings. Similarly, the three groups that received transplants and CSF collections at two different times were grouped into six representative pools for each group and time. Considering the proteomic platform used, the three groups were similarly compared: DPBS After vs. Before, AD-G After vs. Before and BM-G After vs. Before, being found 208, 211 and 131 proteins in each comparison, respectively. Likewise, in the paired comparison of the CSF of the EPM and Control groups, EPM vs. Control, 201 proteins were identified, and 33 proteins were observed exclusively in the CSF of the EPM group. In addition, the presence of the three types of enolases interacting with each other (ENO 1, ENO 2 and ENO 3) was also observed in the comparison of the DPBS, BM-G and EPM groups, being exclusives in DPBS and BM-G at time After and in the EPM group as its exclusively. In this context, this study, when evaluating the proteomic profilings of CSF before and after multiple intrathecal transplantations of MSCs in healthy horses and CSF of horses chronically affected by EPM, allowed the identification of enolases as potential markers of neural and / or EPM injury. Further studies should be performed to confirm these findings and to advance the knowledge of the effects of intrathecal cell therapy with MSCs for the treatment of neurological lesions in horses.

EFEITO DO FLAVONOIDE AGATHISFLAVONA NA MODULAÇÃO DA RESPOSTA GLIAL E NA POPULAÇÃO DE NEURÔNIOS CORTICAIS EM MODELO IN VITRO DE TRAUMATISMO CRANIOENCEFÁLICO

Em um primeiro momento, utilizou-se a prospecção científica para a avaliação ao nível mais alto de desenvolvimento em artigos científicos sobre pesquisa com o flavonoide agathisflavona. A pesquisa prospectiva resultou em 81 produções científicas expressas em diversos periódicos, submetidas por diferentes países, com aumento no número de publicações verificado na última década. Entre as atividades biológicas do flavonoide agathisflavona, já demonstradas, estão efeito hepatoprotetor, antiprotozoario, antibiótico, antiviral, imunomodulador, neurogênico e neuroprotetor. Nesta perspectiva, foi possível investigar os avanços no estudo do flavonoide agathisflavona para desenvolvimento de novas terapêuticas. Em um segundo momento, testou-se os efeitos da agathisflavona em um modelo in vitro de TCE. O TCE é uma patologia multifatorial, sendo causa importante de incapacitação em seres humanos e animais. A depender do estresse mecânico causado pelo TCE, os astrócitos reagem com mudanças complexas de fenótipo e função conhecidas como astrogliose. Em casos de dano tecidual severo, os astrócitos proliferam na área imediatamente adjacente à lesão e se entrelaçam formando a cicatriz glial que, apesar de importante para mitigar o dano inicial do TCE também interfere na posterior regeneração do SNC. O flavonoide agathisflavona tem demonstrado efeito neuroprotetor, neurogênico e imunomodulador, o que pode ser benéfico em situações de TCE. Neste estudo foram investigados os efeitos do flavonoide agathisflavona em um modelo in vitro de TCE em culturas primárias de córtex cerebral embrionário murino em termos de modulação da resposta astrocitária, integridade de neurônios e modulação de fatores neurotróficos e imunomodulatórios. Comparado com os controles, tratamento com agathisflavona (1 µM) diminuiu a expressão de GFAP e espessamento de processos astrocitários observado nas culturas lesionadas, e gerou um aumento no número de corpos de neurônios e crescimento de neuritos que adentram a lesão. Agathisflavona (0,1 e 1 µM) também induziu aumento na expressão de fatores neurotróficos NGF e GDNF, assim como da enzima arginase-1, associada com microglias de perfil neuroprotetor M2. Os resultados encontrados no presente estudo demonstram que o flavonoide agathisflavona modula a resposta astrocitária e formação de cicatriz glial, estimulando a recomposição da rede neural em um modelo in vitro de TCE, o que pode ser considerado para terapia de TCE.

Initially, scientific prospecting was used for evaluation at the highest level of development in scientific papers on research with the flavonoid agathisflavone. The prospective research resulted in 81 scientific productions expressed in several journals, submitted by different countries, with an increase in the number of publications verified in the last decade. Among the biological activities of the flavonoid agathisflavone, already demonstrated, are hepatoprotective, antiprotozoal, antibiotic, antiviral, immunomodulatory, neurogenic and neuroprotective effect. In this perspective, it was possible to investigate the advances in the study of the flavonoid agathisflavone for the development of new therapeutics. In a second step, the effects of agathisflavone on an in vitro model of TBI were tested. ECT is a multifactorial pathology, being an important cause of incapacitation in humans and animals. Depending on the mechanical stress caused by TBI, astrocytes react with complex phenotype and function changes known as astrogliosis. In cases of severe tissue damage, astrocytes proliferate in the area immediately adjacent to the lesion and intertwine forming a glial scar which, while important to mitigate the initial damage of the TBI, also interferes with the subsequent regeneration of the CNS. The flavonoid agathisflavone has demonstrated neuroprotective, neurogenic and immunomodulatory effects, which may be beneficial in situations of TBI. In this study we investigated the effects of flavonoid agathisflavone on an in vitro model of TBI in primary cultures of murine embryonic cerebral cortex in terms of modulation of astrocytic response, neuron integrity and modulation of neurotrophic and immunomodulatory factors. Compared with the controls, treatment with agathisflavone (1 M) decreased GFAP expression and thickening of astrocyte processes observed in the injured cultures, and generated an increase in the number of neuron bodies and neurite outgrowth that entered the lesion. Agathisflavone (0.1 and 1 M) also induced increased expression of NGF and GDNF neurotrophic factors, as well as the arginase-1 enzyme, associated with M2 neuroprotective profile microglia. The results found in the present study demonstrate that the flavonoid agathisflavone modulates the astrocytic response and glial scar formation, stimulating the neural network recomposition in an in vitro model of TBI, which can be considered for TBI therapy.

Sciatic nerve regeneration in rats by a nerve conduit engineering with a membrane derived from natural latex / Regeneração do nervo ciático em ratos através de um conduto confeccionado com uma membrana de látex natural

PURPOSE: To evaluate the capacity of natural latex membrane to accelerate and improve the regeneration quality of the of rat sciatic nerves. METHODS: Forty male adult Wistar rats were used, anesthetized and operated to cut the sciatic nerve and receive an autograft or a conduit made with a membrane derived from natural latex (Hevea brasiliensis). Four or eight weeks after surgery, to investigate motor nerve recovery, we analyzed the neurological function by walking pattern (footprints analysis and computerized treadmill), electrophysiological evaluation and histological analysis of regenerated nerve (autologous nerve graft or tissue cables between the nerve stumps), and anterior tibial and gastrocnemius muscles. RESULTS: All functional and morphological analysis showed that the rats transplanted with latex conduit had a better neurological recovery than those operated with autologous nerve: quality of footprints, performance on treadmill (p<0.01), electrophysiological response (p<0.05), and quality of histological aspects on neural regeneration. CONCLUSION: The data reported showed behavioral and functional recovery in rats implanted with latex conduit for sciatic nerve repair, supporting a complete morphological and physiological regeneration of the nerve.(AU)

OBJETIVO: Avaliar a capacidade de uma membrana de látex natural em acelerar e melhorar a qualidade da regeneração do nervo ciático seccionado de ratos. MÉTODOS: Foram utilizados 40 ratos machos adultos da linhagem Wistar, anestesiados e operados com autoenxerto ou com interposição de um tubo confeccionado com uma membrana derivada do latex natural (Havea brasiliensis). Quatro ou oito semanas após a cirurgia, para investigar a recuperação motora do nervo, foram analisadas a função neurológica através do padrão da marcha (análise das pegadas e esteira computadorizada), avaliação eletrofisiológica e análise histológica do nervo regenerado (enxerto de nervo autólogo ou formação de nervo novo entre os cotos nervosos) e músculos gastrocnêmio e tibial anterior. RESULTADOS: Todas as análises morfológicas e funcionais demonstraram que os ratos transplantados com o conduto de látex tiveram recuperação melhor do que aqueles operados com nervo autólogo: qualidade das pegadas impressas, desempenho em esteira (p<0,01), resposta eletrofisiológica (p<0,05), e qualidade histológica da regeneração nervosa. CONCLUSÃO: Os dados apresentados demonstraram recuperação comportamental e funcional nos ratos implantados com o conduto de látex para a reparação do nervo ciático por meio de uma completa regeneração morfológica e fisiológica do nervo.(AU)

Muscle reinnervation in one or two stages?: experimental study in rats with end-to-side nerve graft / Reinervação muscular em um ou dois estágios?: estudo experimental em ratos com enxerto de nervo término-lateral

PURPOSE: To compare muscle reinnervation in one and two surgical stages using end-to-side neurorrhaphy (ESN) without donor nerve injury. METHODS: The experiment was performed on four groups of 20 rats. Group 1 (G1), one stage, received the graft which was sutured to the tibial nerve, with ESN, and its free stump was sutured end-to-end to the distal stump of the sectioned peroneal nerve (PN), all in the same operation. In Group 2 (G2), two stages, the nerve graft was sutured to the tibial nerve, with ESN. Two months later the PN was sectioned and its distal stump connected to the distal stump of the graft as in G1. Normal control group (Gn) received the graft only sutured to the tibial nerve, with ESN. Denervated control group (Gd), as well received the graft and had the PN sectioned and its two stumps buried in adjacent musculature, with the aim of denervating the cranial tibial muscle (CTM), the target of this study. The parameters used to evaluate CTM reinnervation were muscle mass, muscle fiber's minimum diameter and area. RESULTS: The mean CTM mass, the average of the muscular fibers areas and the average of the muscular fiber minimum diameters was higher (all p<0.0001) in G2 than in G1. Comparing the four groups, these parameters had their maximum expression in Gn and the minimum in Gd, as expected. CONCLUSION: The two stages showed better muscle reinnervation than one stage.(AU)

OBJETIVO: Comparar a reinervação muscular com enxerto de nervo em um e dois tempos operatórios, utilizando a neurorrafia término-lateral (NTL) sem lesão do nervo doador. MÉTODOS: Vinte ratos foram distribuídos em quatro grupos. O grupo 1 (G1), um estágio, recebeu o enxerto que foi suturado ao nervo tibial (NT), por meio de NTL, e seu coto livre foi suturado por NTL ao coto distal do nervo peroneal (NP), seccionado a um centímetro do NT, na mesma cirurgia. O grupo 2 (G2), dois estágios, recebeu o enxerto de nervo na primeira cirurgia, como já descrito. Dois meses depois, na segunda cirurgia, o NP foi seccionado e seu coto distal ligado ao coto distal do enxerto como em G1. O grupo controle de normalidade (Gn) recebeu o enxerto da mesma forma, apenas. E o grupo controle de denervação (Gd), além de receber o enxerto, teve o NP seccionado e seus cotos sepultados na musculatura adjacente, com a finalidade de denervar o músculo tibial cranial (MTC), alvo deste estudo. Os parâmetros utilizados para avaliar a reinervação do MTC foram massa muscular, diâmetro mínimo da fibra muscular e área. RESULTADOS: O grupo G2 apresentou superioridade (p<0,0001) em relação ao G1 na massa do MTC, no diâmetro mínimo e na área das fibras musculares. Na comparação entre os quatro grupos, estes mesmos parâmetros tiveram sua expressão máxima em Gn e mínima em Gd, como era esperado. CONCLUSÃO: A reinervação muscular em dois estágios apresenta melhor resultado quando comparada à técnica em um tempo.(AU)

Isolamento de células derivadas da pele de macacos Sapajus apella (Linnaeus, 1758) e de bovinos para fins de aplicação em biotécnicas da reprodução

Células-tronco presentes nos compartimentos da pele são indispensáveis para a manutenção da homeostase, regeneração, reparação tecidual e reconstrução da integridade e funcionalidade após lesões. Em 2001 um protocolo de isolamento de precursores neurais foi adaptado para o isolamento de células-tronco da pele, referidas como precursores derivados da pele (SKPs), as quais foram diferenciadas, posteriormente, em células da linhagem germinativa, abrindo, assim, uma via de exploração para aplicações em biotecnologias da reprodução. Outro relevante aspecto sobre a pele, são as metodologias desenvolvidas para sua conservação e manutenção por longos períodos, o que permite a estocagem de suprimento biológico para inúmeras pesquisas, inclusive em biologia da conservação e criobiologia. Nesse contexto, considerando-se a necessidade de novas técnicas que contribuam para a preservação de espécies de primatas neotropicais e ungulados silvestres, este trabalho teve como objetivo o isolamento de células derivadas da pele de macacos Sapajus apella e de bovinos, buscando-se, em acréscimo, o estabelecimento de um protocolo de criopreservação das amostras, para posterior utilização em biotécnicas da reprodução. Inicialmente, implementou-se um protocolo otimizado de isolamento enzimático de precursores derivados da pele, com o qual foi possível obter SKPs de fetos bovinos, as quais foram positivas para a atividade da enzima fosfatase alcalina, marcadora de pluripotência. Em Sapajus apella foi possível isolar e cultivar células por essa metodologia até a formação de aglomerados. Em seguida, células resistentes a estresse conhecidas como células Muse, foram isoladas de fibroblastos de macaco Sapajus apella e de bovino adulto e em idade fetal. As células resistentes a estresse formaram aglomerados semelhantes a SKPs em amostras dos três animais, indicando a técnica como um método com potencial para a obtenção de esferas multipotentes. Por fim, estabeleceu-se um protocolo para vitrificação de biópsias de pele de macacos Sapajus apella, o qual foi efetivo para manter a integridade de até 88% das células epidermais, preservar boa densidade de fibroblastos e viabilizar a obtenção de células através do cultivo primário por explante. O protocolo de vitrificação proposto nesse estudo, associado às técnicas de isolamento de células derivadas da pele executadas, constituem em valiosa metodologia para a formação de bancos de germoplasma que viabilizem a realização de biotécnicas da reprodução como a clonagem e a geração de gametas in vitro e contribuam para a preservação de espécies silvestres em risco de extinção.

Stem cells present in the skin compartments are indispensable for the maintenance of homeostasis, regeneration, tissue integrity repair and functionality recovering after injury. In 2001 a protocol for the isolation of neural precursors was adapted for the isolation of skin stem cells, referred as "skin derived precursors" (SKPs), which were able to differentiate into germline cells, thus opening up a line of research in the field of reproduction biotechnology.Another relevant aspect about the skin are the methodologies developed for its conservation and maintenance for long periods, which allows the storage of biological supplies for numerous researches, including conservation biology and cryobiology. In this context, considering the need for new techniques that contribute to the preservation of neotropical primates and wild ungulates species, this work aimed to isolate skin derived cells of Sapajus apella monkeys and bovines. In addition, it was stablished a cryopreservation protocol for the skin samples. Initially, an optimized protocol of enzymatic isolation of skin derived precursors was implemented. It was possible to obtain alkaline phosphatase positive SKPs from bovine fetuses and to isolate and cultures cells from Sapajus apella until cluster formation. Next, stress-tolerant cells also known as Muse cells, were isolated from fibroblastsof Sapajus apella, adult bovine and bovine fetus. The stress-tolerant cells isolated from all three animals generated SKP-like spheres, indicating the technique as a potential method for multipotent spheres obtention. Finally, a protocol for Sapajus apella skin biopsies vitrification was established, which was effective to maintain the integrity of up to 88% of epidermal cells, preserve good fibroblast density and to be established a primary culture of skin explants.The vitrification protocol proposed in this study, associated with the techniques of skin derived cells isolation, constitute a valuable methodology for the formation of germplasm banks that enable the performance of reproductive biotechniques such as cloning and in vitro gametes generation and contribute to the preservation of endangered or threatened wild species.

Histological analysis of low-intensity laser therapy effects in peripheral nerve regeneration in Wistar rats / Avaliacao histologica dos efeitos da laserterapia de baixa potencia sobre os processos de regeneracao nervosa periferica em ratos Wistar

Purpose: Analyze the influence of low-intensity laser therapy in the sciatic nerve regeneration of rats submitted to controlled crush through histological analysis. Methods: Were used 20 Wistar rats, to analyze the influence of low-intensity laser therapy in the sciatic nerve regeneration, where the injury of the type axonotmesis was induced by a haemostatic clamp Crile (2nd level of the rack). The animals were randomly distributed in 2 groups. Control group (CG n = 10) and Laser group (LG n = 10). These were subdivided in 2 subgroups each, according to the euthanasia period: (CG14 _ n = 5 and CG21 _ n = 5) and (LG14 _ n = 5 and LG21 _ n = 5). At the end of treatment, the samples were removed and prepared for histological analysis, where were analyzed and quantified the following findings: Schwann cells, myelinic axons with large diameter and neurons. Results: In the groups submitted to low-intensity laser therapy, were observed an increase in the number of all analyzed aspects with significance level. Conclusion: The irradiation with low intensity laser (904nm) influenced positively the regeneration of the sciatic nerve in Wistar rats after being injured by crush (axonotmesis), becoming the nerve recovery more rapid and efficient.(AU)

Objetivo: Verificar a influencia da terapia com laser de baixa potencia na regeneracao histologica do nervo ciatico de ratos submetidos a neuropraxia controlada. Metodos: Foi utilizada a amostra de 20 ratos da linhagem Wistar, para verificar a influencia da terapia com laser de baixa intensidade na regeneracao nervosa periferica, onde a lesao do tipo axoniotmese foi induzida por meio de preensao com pinca hemostatica de Crile. Os animais foram distribuidos randomicamente dois grupos. Grupo controle (CG n = 10), e Grupo laser (LG n = 10). Cada um destes grupos foi subdividido em dois subgrupos dependendo do periodo da eutanasia: (CG14 - n = 5 e CG21 - n = 5) e (LG14 - n = 5 e LG21 - n = 5). Ao final do tratamento, amostras do nervo foram retiradas e analisadas histologicamente, nas quais foi adotado na pesquisa a analise do numero de neuronios, de celulas de Schwann (CS) e de axonios mielinicos de grande diametro. Resultados: Nos grupos submetidos a terapia com laser de baixa potencia foi observado aumento do numero de todos os aspectos analisados com diferenca estatisticamente significante. Conclusao: A irradiacao com o laser de baixa intensidade (904nm) influenciou positivamente na regeneracao do nervo ciatico de ratos da linhagem Wistar pos neuropraxia controlada (axonotmese), tornando a recuperacao nervosa mais rapida e eficiente.(AU)

End-to-side nerve repair using fibrin glue in rats / Anastomose nervosa término-lateral usando cola de fibrina em ratos

Purpose: To evaluate axonal regeneration after end-to-side nerve repair with fibrin glue in rats. Methods: Forty-five Wistar rats were divided into three groups: group A (n=15), were not submitted to surgery (control group); group B (n=15) were submitted to fibular transection without repair; and group C (n=15), were submitted to fibular transection with end-to-side nerve anastomosis using fibrin glue, in the lateral surface of an intact tibial nerve. The three groups were submitted to walking track (30 and 90 days) and posterior morphometrical analysis (90 days). Results: The functional tests demonstrated that there was no difference in the walking track during the study in group A (p>0.05). The group B had walking pattern impairment in the two tests (p>0.05). The group C had walking pattern impairment in the first test, with important recovery in the second test (p<0.05). The morphometrical assessment revealed significantly higher number of regenerated mielinates axons in group C, compared to group B (p<0.05). Conclusion: The end-to-side nerve repair with fibrin glue shows axonal recovery, demonstrated through functional and morphometrical ways in rats.(AU)

Objetivo: Avaliar a regeneração axonal após anastomose nervosa término-lateral (ATL) usando cola de fibrina em ratos. Métodos: Quarenta e cinco ratos Wistar distribuídos em três grupos: os animais do grupo A (n=15) não foram submetidos à secção nervosa (grupo controle); os animais do grupo B (n=15) foram submetidos apenas à secção do nervo fibular, sem posterior anastomose; e os animais do grupo C (n=15) foram submetidos à secção do nervo fibular e à ATL com cola de fibrina no nervo tibial. Posteriormente, os animais foram submetidos a dois testes de marcha (30 e 90 dias) e à análise morfométrica (90 dias). Resultados: A análise estatística dos testes de marcha demonstrou que o grupo A não apresentou alteração no padrão de caminhada durante o estudo (p>0,05). O grupo B apresentou prejuízo motor no primeiro e no segundo teste (p>0,05). O grupo C apresentou um padrão de atrofia no primeiro teste, com recuperação da marcha no segundo teste (p<0,05). Na análise morfométrica, o grupo C apresentou regeneração axonal significativamente superior ao grupo B (p<0,05). Conclusão: A ATL realizada com cola de fibrina resultou em regeneração axonal no rato, demonstrada tanto histologicamente quanto funcionalmente.(AU)

Terapia celular aplicada a lesão medular induzida em coelhos (Oryctolagus cuniculus)

A causa mais comum de lesão da medula espinhal são os traumas de alto impacto, que frequentemente resultam em algum comprometimento motor, sensitivo ou autonômico em maior ou menor grau nas áreas distais ao nível do trauma. Em termos de sobrevivência e complicações devido a sequelas, os pacientes veterinários têm um prognóstico reservado a desfavorável. Portanto justifica-se o estudo de modelos experimentais de produção de lesão medular espinhal e posterior tratamento com terapia celular, que possam prover mais subsídios para pesquisa no âmbito clínico na medicina e medicina veterinária. Estudos pré-clínicos de lesão medular espinhal aguda requerem um modelo experimental animal reprodutível facilmente. O objetivo da tese foi avaliação do possível grau de regeneração neural e recuperação funcional de coelhos submetidos experimentalmente a lesão medular espinhal provocada pela técnica de compressão medular e tratados com células tronco mesenquimais ou células mononucleares derivadas de medula óssea, em diferentes tempos e por conseguinte comparar a terapia com estes dois tipos celulares, utilizando coelhos como modelo experimental por se tratar de uma espécie que apresenta um número menor de publicações conclusivas e que contempla os principais requisitos de um modelo como, baixo custo, praticidade de manejo, reprodutibilidade e uniformidade. Concluímos que o tratamento deste tipo de lesão com as terapias celulares propostas são benéficas na recuperação clínica dos animais.

The most common cause of spinal cord injury are high impact trauma, which often result in some motor impairment, sensory or autonomic a greater or lesser extent in the distal areas the level of trauma. In terms of survival and complications due to sequelae, veterinary patients have a poor prognosis unfavorable. Therefore justified the study of experimental production spinal cord injury and subsequent treatment with cell therapy, which can provide more support for research in the clinical setting in medicine and veterinary medicine. Preclinical studies of acute spinal cord injury require an experimental animal model easily reproducible. The purpose of these evaluation was possible degree of neural regeneration and functional recovery of experimental rabbits underwent spinal cord injury caused by spinal compression technique and treated with mesenchymal stem cells or the mononuclear cells derived from bone marrow, at different times and therefore compare therapy with these two cell types, using rabbits as an experimental model because it is a species that has fewer conclusive publications and contemplating the main requirements of a model as, low cost, management convenience, reproducibility and uniformity. We conclude that treatment of this type of injury with cellular therapies proposals are beneficial in clinical recovery of the animals

Neurorrafia do ramo bucal dorsal do nervo facial em coelhos com proteção de segmento intestinal alógeno / Neurorrhaphy of the dorsal buccal branch of facial nerve in rabbits with protection o f allograft intestinal segment

Foram utilizados 18 coelhos, Nova Zelândia, machos, adultos, para avaliação clínica e histológica do reparo do ramo bucal dorsal do nervo facial, decorridos 15, 30 e 60 dias de pós-operatório (PO). Os animais foram distribuídos em dois grupos para secção e aproximação epineural do ramo bucal com fio náilon monofilamentoso 10-0. Nos animais do grupo I, o nervo foi revestido com proteção de segmento de jejuno alógeno conservado em glicerina a 98% e o grupo II apenas aplicação de sutura epineural. Nos coelhos dos dois grupos ocorreu retorno da movimentação do lábio superior a partir da oitava semana.Verificou-se infiltrados celulares e células gigantes com fibrose desorganizada e fibras colágenas do envoltório alógeno entremeadas ao tecido conjuntivo. Aos 15 e 30 dias de PO, os cotos distais deambos os grupos encontravam-se com degeneração walleriana e aos 60 dias, com fibras regeneradas. A reparação do ramo bucal dorsal do nervo facial com o segmento intestinal não foi significativamente diferente nos coelhos do controle, quanto à avaliação de recuperação funcional e histológica.(AU)

18 rabbits, New Zealand, males, adults were used for clinical and histological evaluation of repair dorsal buccal branch of facial nerve after 15, 30 and 60 days postoperatively (PO). The animals were divided into two groups for transection and 10-0 nylon monofilament epineural suture of buccal branch. In animals in Group I, the nerve was coated with protection of jejunum allograft preserved in glycerin 98% and in group II was applied epineural suture. Both groups occurred the return of movement of the upper lip from the eighth week. There was infiltrated cellular and giant cells with fibrosis unsystematic and collagen fibers of the allograft jejunum joing to the connective tissue. At 15 and 30 days of PO, the distal nerve stumps of both groups were found with degeneration wallerian and in 60 days, regenerated fibers. The repair of the dorsal buccal branch offacial nerve with the allograft wasnt significantly different between the control rabbits as to the assessment of histological and functional recovery.(AU)

Histopathological analysis of gangliosides use in peripheral nerve regeneration after axonotmesis in rats / Análise histopatológica da regeneração nervosa periférica com uso de gangliosídeos após axonotmese em ratos

PURPOSE: To analyze the action of gangliosides in peripheral nerve regeneration in the sciatic nerve of the rat. METHODS: The sample was composed of 96 male Wistar rats. The animals were anaesthetized and, after identification of the anaesthesic plane, an incision was made in the posterior region of the thigh, followed by skin and muscle divulsion. The right sciatic nerve was isolated and compressed for 2 minutes. Continuous suture of the skin was performed. The animals were randomly divided into two groups: the experimental group (EG), which received subcutaneous injection of gangliosides, and the control group (CG), which received saline solution (0.9 percent) to mimic the effects of drug administration. RESULTS: No differences were observed between the experimental and control groups evaluated on the eighth day of observation. At 15 and 30 days the EG showed an decrease in Schwann cell activity and an apparent improvement in fibre organization; at 60 days, there was a slight presence of Schwann cells in the endoneural space and the fibres were organized, indicating nerve regeneration. At 15 and 30 days, the level of cell reaction in the CG had diminished, but there were many cells with cytoplasm in activity and in mitosis; at 60 days, hyperplastic Schwann cells and mitotic activity were again observed, as well as nerve regeneration, but to a lesser extent than in the EG. CONCLUSION: The administration of exogenous gangliosides seems to improve nerve regeneration.(AU)

OBJETIVO: Avaliar os efeitos de gangliosídeos na regeneração nervosa periférica em nervo isquiático de ratos após axonotmese. MÉTODOS: Foram utilizados 96 ratos machos albinos (Wistar). Os animais foram anestesiados e após constatação do plano anestésico, foi realizada incisão na face posterior da coxa direita do animal. Em seguida, foi realizada a dissecção cirúrgica da pele e do músculo e divulsão dos músculos. O nervo isquiático direito foi isolado e sofreu compressão por 2 minutos. Efetuou-se a sutura contínua da pele. Os animais foram distribuídos aleatoriamente em 2 grupos: experimental (GE) que receberam gangliosídeos pela via sub-cutânea e controle (GC) que receberam soro fisiológico 0,9 por cento com a finalidade de mimetizar os efeitos de administração da droga de estudo. A análise histopatológica foi realizada em 8, 15, 30 e 60 dias. RESULTADOS: Não se evidenciaram diferenças significantes entre os grupos controle e experimental avaliados com 8 dias. Nos grupos experimentais de 15 e 30 dias observou-se uma diminuição da atividade das células de Schwann e aparente melhora na organização das fibras nervosas; com 60 dias havia discreta presença células de Schwann no espaço endoneural e as fibras nervosas estavam organizadas sinalizando a regeneração nervosa. Nos grupos controles de 15 e 30 dias o padrão de reação celular diminuiu, entretanto havia muitas células com citoplasmas em atividade e mistose; com 60 dias observou-se ainda a presença de hiperplasia de células de Schwann, atividade mitótica ainda presente e regeneração nervosa presente, porém em menor grau comparando-se com aquele visto no grupo experimental. CONCLUSÃO: A administração de gangliosídeos exógenos parece incrementar a regeneração nervosa.(AU)

Utilização de células-tronco autólogas de medula óssea na regeneração do nervo tibial de coelhos mediante técnica de tubulização com prótese de silicone

This study presents an experimental model of an acute deffect in a peripheral nerve to evaluate neural regeneration using a tubulization technique associated with the inoculation of autologous stem cells from bone marrow. A total of 12 New Zealand white rabbits underwent a bilateral dissection of the tibial nerve followed by repair with silicone tubulization. On the left tibial nerve of all animals, the tube was filled with autologous bone marrow-derived stem cells collected from the humerus. For control, using the same repair technique, the tubes were filled with a NaCl solution in the right tibial nerve. After 30 days of observation, the animals were euthanized and a histological evaluation of the collected nerve segments was performed by staining with hematoxylin-eosin, luxol fast blue, and toluidine blue. From the results it is possible to conclude that the transplanted autologous stem cells associated with the tubulization technique present an advantage in the peripheral nerve regeneration process.

Neste estudo é apresentado um modelo experimental de defeito agudo em nervo periférico para avaliação da regeneração nervosa mediante técnica de tubulização associada à inoculação de células-tronco autólogas de medula óssea. Foram utilizados 12 coelhos Nova Zelândia albinos, submetidos à secção bilateral e ao afastamento de 5mm do nervo tibial e posterior reparo mediante utilização de câmara de silicone. Internamente à prótese de tubulização do nervo tibial esquerdo em todos os animais, foram inoculadas células-tronco autólogas de medula óssea, coletadas a partir do úmero. Como grupo controle (nervo tibial direito), mediante aplicação da mesma técnica de reparo, solução de NaCl 0,9% foi administrada internamente à prótese. Após 30 dias de observação, os animais foram eutanasiados e foi realizada a avaliação histológica dos segmentos nervosos por meio das colorações de hematoxilina-eosina, luxol fast blue e azul de toluidina. Com os resultados, foi possível concluir que o transplante de células-tronco autólogas associado à técnica de tubulização apresenta vantagens no processo de regeneração nervosa periférica.

Utilização de células-tronco autólogas de medula óssea na regeneração do nervo tibial de coelhos mediante técnica de tubulização com prótese de silicone

This study presents an experimental model of an acute deffect in a peripheral nerve to evaluate neural regeneration using a tubulization technique associated with the inoculation of autologous stem cells from bone marrow. A total of 12 New Zealand white rabbits underwent a bilateral dissection of the tibial nerve followed by repair with silicone tubulization. On the left tibial nerve of all animals, the tube was filled with autologous bone marrow-derived stem cells collected from the humerus. For control, using the same repair technique, the tubes were filled with a NaCl solution in the right tibial nerve. After 30 days of observation, the animals were euthanized and a histological evaluation of the collected nerve segments was performed by staining with hematoxylin-eosin, luxol fast blue, and toluidine blue. From the results it is possible to conclude that the transplanted autologous stem cells associated with the tubulization technique present an advantage in the peripheral nerve regeneration process.

Neste estudo é apresentado um modelo experimental de defeito agudo em nervo periférico para avaliação da regeneração nervosa mediante técnica de tubulização associada à inoculação de células-tronco autólogas de medula óssea. Foram utilizados 12 coelhos Nova Zelândia albinos, submetidos à secção bilateral e ao afastamento de 5mm do nervo tibial e posterior reparo mediante utilização de câmara de silicone. Internamente à prótese de tubulização do nervo tibial esquerdo em todos os animais, foram inoculadas células-tronco autólogas de medula óssea, coletadas a partir do úmero. Como grupo controle (nervo tibial direito), mediante aplicação da mesma técnica de reparo, solução de NaCl 0,9% foi administrada internamente à prótese. Após 30 dias de observação, os animais foram eutanasiados e foi realizada a avaliação histológica dos segmentos nervosos por meio das colorações de hematoxilina-eosina, luxol fast blue e azul de toluidina. Com os resultados, foi possível concluir que o transplante de células-tronco autólogas associado à técnica de tubulização apresenta vantagens no processo de regeneração nervosa periférica.