Transplantaments glials combinats amb neuroprotecció i restitució neuronal per a la reparació de lesions per avulsió d'arrels espinals

This Project aims to ascertain if combined treatment by surgical reimplantation of the lesioned roots together with local transplant of ensheathing glía and administration of neuroprotective drugs, enhances survival and axonal regeneration of spinal motoneurons and of sensory afferents after severe injuries by avulsion of spinal roots. By means of an adequate combination of therapies we will attempt to improve neuronal survival after the avulsion and to achieve an effective functional recovery, in order to solve the three main consequences of root and plexus lesions: paralysis, anesthesia and neuropathic pain. In addition, the restitution of motoneuronas lost after injury will be attempted by delayed transplantation of predetermined stem cells. In the project an in vivo model of lumbar root avulsion in the adult rat will be used. First, the technique and time window most adequate for the reimplantation repair will be determined. Using an in vitro model of spinal cord slice culture, neuroprotective agents will be assessed, for selecting the most effective for administration in rats after root avulsion. Ensheathing glial cells will be cultured, and transplanted in situ at the zone of root reimplantation, as coadjuvant treatment to favour axonal regeneration, particularly in cases of delayed reimplantation. Embryonic stem cells will be predetermined in culture towards expressing a motoneuronal phenotype. Such cells will be transplanted into the lumbar spinal cord of rats with root avulsion, in order to evaluate if they differentiate and allow for restitution of dead motoneuronas. The animals will be evaluates at several intervals postlesiona by using functional (locomotion, algesimetry) and electrophysiological methods (nerve conduction, evoked potentials). At the end of follow-up morphological and immubohistochemical techniques will be used to characterize neuronal and glial responses and axonal regeneration