Previous studies have suggested that sympathetic sprouting in the periphery may contribute to the development and persistence of sympathetically-maintained pain in animal models of neuropathic pain. The purpose of this thesis was to examine morphological changes in the cutaneous innervation in rats after chronic constriction injury (CCI) to the sciatic nerve. More specifically, this study addresses the question of whether sympathetic fibres sprout de novo into the upper dermis of the rat hindpaw skin after CCI of the sciatic nerve. We also determined changes in peptidergic sensory innervation following CCI. / At several periods post-injury, hind paw skin was harvested and processed using a monoclonal antibody against dopamine-beta-hydroxylase to detect sympathetic fibres and a polyclonal antibody against calcitonin gene-related peptide to identify peptidergic sensory fibres. We observed migration and branching of sympathetic fibres into the upper dermis of the hind paw skin, from where they were normally absent. This migration was first detected at 2 weeks, peaked at 4 to 6 weeks and lasted for at least 20 weeks post-lesion. At 8 weeks post-lesion, there was a dramatic increase in the density of peptidergic fibres in the upper dermis. Quantification revealed that densities of peptidergic fibres 8 weeks post-lesion were significantly above levels of sham animals. Interestingly, the ectopic sympathetic fibres did not innervate blood vessels but formed a novel association and wrapped around sprouted peptidergic nociceptive fibres. Our data show a long-term sympathetic and sensory innervation change in the rat hind paw skin after the chronic constriction injury. This novel fibre arrangement after nerve lesion may play an important role in the development and persistence of sympathetically-maintained neuropathic pain after partial nerve lesions.

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Orientador: Marcus Alexandre Finzi Corat / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Made available in DSpace on 2018-08-24T17:01:17Z (GMT). No. of bitstreams: 1 Nicolielo_RenatoLuizCursino_D.pdf: 3694566 bytes, checksum: 9d48540bdf4e330a06198c2b7816ad32 (MD5) Previous issue date: 2014 / Resumo: As lesões traumáticas de nervos periféricos têm sido objeto de numerosos estudos que tentam compreender os processos de reparação pós-lesão. Apesar dos avanços nas técnicas cirúrgicas, os resultados funcionais têm sido bastante decepcionantes, ao que se refere ao intervalo de tempo entre lesão e tratamento. Para tratamento rápido e eficaz de lesões nervosas, a terapia celular torna-se uma possibilidade no aumento do sucesso de reparo do nervo. Propõe-se então avaliar a eficácia da terapia celular, sobre os níveis de melhora funcional dos camundongos com o nervo isquiático lesionado e tratado com células progenitoras GFP da medula óssea em comparação com seus controles. No presente trabalho utilizou-se camundongos C57BL/6-HETGFP/J/Unib geneticamente modificados como doadores de células progenitoras GFP ("green fluoresent protein") da medula óssea que foram usadas no tratamento via sistêmica em camundongos C57BL/6/J/Unib não marcados com GFP submetidos a esmagamento do nervo isquiático. Estes animais foram analisados quanto sua performance funcional, migração celular, imunohistoquímica de proteínas p75NTR e neurofilamento e contagem axonal em imagens de microscopia eletrônica. Através dos resultados obtidos verificou-se que houve uma significativa melhora funcional do grupo tratado de 224,9% quando comparado com o grupo controle, principalmente no início do processo de recuperação. Estes valores de melhora funcional coincidiram com a grande quantidade de células GFP, maior quantidade de axônios mielinizados e menor quantidade de axônios degeneradas encontradas no sitio da lesão, além do aumento de imunorreatividade da proteína p75NTR, envolvida no processo de regeneração nervosa. Tudo isto vem a demonstrar a eficiência da terapia celular utilizada com células progenitoras da medula óssea, com uma purificação não especifica, tornando-a viável para uma terapia celular sistêmica rápida, simples e eficaz sendo assim uma possibilidade terapêutica para lesões de nervo / Abstract: Traumatic injuries of peripheral nerves have been the subject of numerous studies that attempt to understand the processes of repair after injury. Despite advances in surgical techniques, the functional results have been very disappointing to depend on the time interval between injury and treatment. For fast and effective treatment of nerve lesions, cell therapy becomes a possibility in increasing the success of nerve repair. So it is proposed to evaluate the efficacy of cell therapy on levels of functional improvement of mice with injured ischiatic nerve and treated with GFP progenitor cells of bone marrow as compared with their controls. In this study we used mices C57BL/6-HETGFP/J/Unib genetically modified as donors of stem cells GFP ("green fluoresent protein") of bone marrow that were used in the treatment via systemic of mice C57BL/6/J/Unib not marked with GFP underwent sciatic nerve crush. These animals were analyzed for their functional performance, cell migration, immunohistochemistry of p75NTR protein and neurofilament and axonal counts in electron microscope images. From the results obtained it was verified that there was a significant functional improvement in the treated group 224.9% when compared with the control group, mainly at the start of the recovery process. These values of functional improvement have coincided with the large amount of GFP cells, largest amount of myelinated axons and smallest amount of degenerate axons found at the site of injury axons, in addition to the increase of immunoreactivity of the protein of p75NTR involved in nerve regeneration process. All this is to demonstrate the efficiency of cell therapy used in bone marrow progenitor cells with a non-specific purification, making it feasible for a quick, simple and effective systemic cell therapy becoming this way a therapeutic possibility for nerve injured / Doutorado / Clinica Medica / Doutor em Clínica Médica