Among the neurodegenerative diseases of the central nervous system, multiple sclerosis (MS) is a chronic inflammatory disease represents the major cause of non-traumatic neurological disability in young adults. Current treatments are based on the use of immunosuppressants for controlling inflammation in early stages. These treatments have been shown to reduce rates of recurrence and disease activity, but have had a modest impact on the progression of disease and disability associated, especially in aggressive forms. Animal models of MS have recently allowed studying the effect of hMSCs (human mesenchymal cells) after intravenous infusion and demonstrated a decrease in the degree of injury, reduced short-term functional deficits and, what is most striking , an improvement in long-term recovery following an immunomodulatory effect, associated with a neuroprotective potential (reduction in astrogliosis and increased number of oligodendrocytes and neurons). In the human adult brain there is limited capacity for axonal regeneration. This is due to multiple converging mechanisms among which are the inhibition of axonal growth, inhibition of endogenous neural stem cells and overexpression of inhibitory molecules by reactive cells at sites of injury. Once you have formed the "astroglial scar", little can be done because it has formed a biochemical and physical barrier for repair. One type of molecules that inhibit axonal regeneration and that are associated with myelin (Myelin-English MAIs Associated Inhibitors). Three of them have been extensively studied: Nogo-A, MAG and OMgp. All converge on the Nogo receptor (NgR1), it has shown that the role of Nogo in MS is to prevent functional recovery from injury by inhibition of axonal growth. Considering the background mentioned above, the hMSCs offer us the theoretical possibility of getting two effects, immunomodulatory and neuroprotective. These effects, along with strategies to act locally on the axonal repair (lock NogoA), could result in a clinically significant benefit for patients with MS. HYPOTHESES AND OBJECTIVES Based on the knowledge that hMSCs have tropism for inflamed regions and neurorregenerador role of Nogo receptor blockade, we propose the following hypotheses: 1. There MSC tropism of the inflammatory microenvironment optic nerve animal model of multiple sclerosis. 2. Due to this tropism of MSCs may neuroprotective action at the ocular level, resulting histological improvement in optic neuritis. Our objectives are: 1. Study of the inflammatory cellular microenvironment of the optic nerve and retina in the animal model of MS. 2. Study of the neuroprotective and neuroregenerative response to the optic nerve and retina of human mesenchymal stem cells (hMSCs) transfected with a gene neurorregenerador (Nogo receptor negative dominant form). MATERIAL AND METHODS In this thesis we have generated an animal model of MS, a model of EAE (experimental autoimmune encephalitis) with myelin protein (MOG) in mice B6 / C57, after immunization we have studied the changes that occur in the retina and nerve optical (NO) and have analyzed the effect of intravenous infusion of mesenchymal cells transfected with a specific gene (MSC-NogoRDN) to promote axonal regeneration. EAE animal model were monitored and followed by motor disability scales. Eyeballs, retinas and optic nerves were obtained and processed for: 1) techniques morphological and morphometric examination using light microscopy, 2) immunofluorescence techniques, 3) techniques to electronic microspopía ultrastructural examination. RESULTS With the results of the monitoring of the animals in both groups immunized (MOG vs MOG+MSC-NogoRDN) animals receiving treatment, they had a free period increased illness and had less disability than untreated animals. The results showed an improvement in the parameters analyzed in NO and retina after administration of MSCs transfected. In the case of the retina, it decreased RGC loss (although it was more a trend that did not reach statistical significance) and prevented the loss of myelin by 34% and reduced the degree of inflammation (as gliosis and cellularity inflammatory), the latter with p = 0.002. CONCLUSIONS Based on the results obtained in the experiments described in this thesis, in the mouse EAE model generated B6 / C57, by MOG we conclude: 1.-The inflammatory microenvironment, which had generated, which had a significant alteration layer morphological and quantitative CGR; and he was not affected with a reactive gliosis and increased axonal damage. 2. We found a neuroprotective effect after administration of MSCs transfected with NogoRDN, lower appreciate RGC loss decreased loss of myelin and lesser degree of inflammation in the NO. 3. We did not find a regenerative effect due to blockade of Nogo, did not succeed in reversing the axonal damage. 4. Confirm tropism hMSCs to inflamed NO, after intravenous inoculation tissue.
2016-02-04
