Safety and seizure control in patients with mesial temporal lobe epilepsy treated with regional superselective intra-arterial injection of autologous bone marrow mononuclear cells.

Temporal lobe epilepsy (TLE) is a highly prevalent syndrome among people with epilepsy, and is usually refractory to drug treatment. Structural and physiological changes, such as hippocampal sclerosis, are often present in TLE patients. The objective of this study is to evaluate the feasibility and safety of intra-arterial infusion of autologous bone marrow mononuclear cells (BMMC) in adults with medically refractory mesial TLE (MTLE) and unilateral hippocampal sclerosis (HS). We enrolled 20 patients who had been diagnosed with MTLE-HS and were refractory to medical treatment. All patients underwent a neurological evaluation, magnetic resonance imaging with hippocampal volumetry, video-electroencephalography (EEG) with ictal recording, and a neuropsychological test battery focusing on verbal and nonverbal memory domains. After bone marrow aspiration and subsequent cell preparation, the BMMC were infused by selective posterior cerebral artery catheterization. Patients were followed for 6 months. Safety of the procedure, seizure frequency, neuropsychological evaluation, EEG variables, routine brain magnetic resonance imaging and hippocampal volumetry were considered measurements of outcome. Any serious intercurrent clinical event or adverse effects related to the procedure were reported. No additional lesions and no significant hippocampal volumetric changes were observed. EEG recordings showed a decrease in theta activity and spike density. At 6 months, eight patients (40%) were seizure free. A significant increase in the memory scores over time was observed. The BMMC autologous transplant for the treatment of temporal lobe epilepsy is feasible and safe. The seizure control achieved in this novel study supports the therapeutic potential of stem cell transplants in MTLE-HS patients. Copyright © 2016 John Wiley & Sons, Ltd.

Morphofunctional analysis of sciatic nerve and motor performance of rats after cryotherapy with liquid nitrogen.

OBJECTIVE: This work evaluated sciatic nerve regeneration after cryotherapy. STUDY DESIGN: Rats underwent surgical access of the sciatic nerve and subsequent cryotherapy, crush lesion, or no manipulation. Walking-track, electroneuromyographic, and histomorphometric analyses were performed at 15, 30, and 70 postoperative days. RESULTS: At 15 days, the crush and cryotherapy groups showed significant morphofunctional impairment. At 30 days, functional loss was significant in the walking-track, but at 70 days, there were no significant differences between the groups. Amplitude was near zero for the crush group at 15 and 30 days and zero for the cryotherapy group. Measurement of latency was not possible in the latter group. Crush and cryotherapy groups showed greater amounts of myelinated fibers (by 30 days), with axonal diameter and width of the myelin sheath being less than in control group. CONCLUSIONS: Sciatic nerve lesion by application of liquid nitrogen is classified as axonotmesis, which is reversible.

Transplantation of bone marrow mononuclear cells decreases seizure incidence, mitigates neuronal loss and modulates pro-inflammatory cytokine production in epileptic rats.

Approximately 30% of patients with mesial temporal lobe epilepsy do not respond to treatment with antiepileptic drugs. We have previously shown that transplantation of mononuclear bone marrow cells (BMC) has an anticonvulsant effect in acute epilepsy. Here, we used pilocarpine to induce epilepsy in rats and studied the effects of BMC injected intravenously either at the onset of seizures or after 10 months of recurrent seizures. BMC effectively decreased seizure frequency and duration. In addition, decreased levels of proinflammatory cytokines (TNF-α, IL-1ß and IL-6) and increased levels of anti-inflammatory cytokine (IL-10) were observed in the brain and serum of BMC-treated rats. Transplants performed at seizure-onset protected against pilocarpine-induced neuronal loss and gliosis and stimulated the proliferation of new neurons in epileptic rats. Our data demonstrate that BMC transplantation has potent therapeutic effects and could be a potential therapy for clinically intractable epilepsies.

NAP prevents acute cerebral oxidative stress and protects against long-term brain injury and cognitive impairment in a model of neonatal hypoxia-ischemia.

Hypoxia-ischemia (HI) is a common cause of neonatal brain damage with lifelong morbidities in which current therapies are limited. In this study, we investigated the effect of neuropeptide NAP (NAPVSIPQ) on early cerebral oxidative stress, long-term neurological function and brain injury after neonatal HI. Seven-day-old rat pups were subjected to an HI model by applying a unilateral carotid artery occlusion and systemic hypoxia. The animals were randomly assigned to groups receiving an intraperitoneal injection of NAP (3 µg/g) or vehicle immediately (0 h) and 24 h after HI. Brain DNA damage, lipid peroxidation and reduced glutathione (GSH) content were determined 24 h after the last NAP injection. Cognitive impairment was assessed on postnatal day 60 using the spatial version of the Morris water maze learning task. Next, the animals were euthanized to assess the cerebral hemispheric volume using the Cavalieri principle associated with the counting point method. We observed that NAP prevented the acute HI-induced DNA and lipid membrane damage and also recovered the GSH levels in the injured hemisphere of the HI rat pups. Further, NAP was able to prevent impairments in learning and long-term spatial memory and to significantly reduce brain damage up to 7 weeks following the neonatal HI injury. Our findings demonstrate that NAP confers potent neuroprotection from acute brain oxidative stress, long-term cognitive impairment and brain lesions induced by neonatal HI through, at least in part, the modulation of the glutathione-mediated antioxidant system.

Prevention of seizures and reorganization of hippocampal functions by transplantation of bone marrow cells in the acute phase of experimental epilepsy.

In this study, we investigated the therapeutic potential of bone marrow mononuclear cells (BMCs) in a model of epilepsy induced by pilocarpine in rats. BMCs obtained from green fluorescent protein (GFP) transgenic mice or rats were transplanted intravenously after induction of status epilepticus (SE). Spontaneous recurrent seizures (SRS) were monitored using Racine's seizure severity scale. All of the rats in the saline-treated epileptic control group developed SRS, whereas none of the BMC-treated epileptic animals had seizures in the short term (15 days after transplantation), regardless of the BMC source. Over the long-term chronic phase (120 days after transplantation), only 25% of BMC-treated epileptic animals had seizures, but with a lower frequency and duration compared to the epileptic control group. The density of hippocampal neurons in the brains of animals treated with BMCs was markedly preserved. At hippocampal Schaeffer collateral-CA1 synapses, long-term potentiation was preserved in BMC-transplanted rats compared to epileptic controls. The donor-derived GFP(+) cells were rarely found in the brains of transplanted epileptic rats. In conclusion, treatment with BMCs can prevent the development of chronic seizures, reduce neuronal loss, and influence the reorganization of the hippocampal neuronal network.

NAP prevents hippocampal oxidative damage in neonatal rats subjected to hypoxia-induced seizures.

Neonatal seizures in which hypoxic-ischemic encephalopathy is the main triggering etiology have a challenging diagnosis and limited efficacy of treatment. NAP (NAPVSIPQ) has shown extensive neuroprotective and antioxidant capacity in vitro and in vivo. To evaluate its neuroprotective role in the context of seizures associated with perinatal hypoxia, we assessed the integrity of DNA and lipid membranes as well as the redox status in the hippocampus of 10-day-old rats exposed to hypoxia-induced seizures (HS) with and without NAP treatment. Rats were exposed to transient global hypoxia (12 min exposure to 5-7% O2 was able to induce electrographic seizures) or room air with subsequent intraperitoneal NAP (0.03, 0.3 or 3 microg/g) or vehicle administration. Results showed elevated DNA damage immediately after the insult until 72 h post-HS, while oxidized bases were only detected 3, 6 and 24 h later. In addition, thiobarbituric acid reactive species peaked at 6 h in parallel with decreased levels of reduced glutathione between 3 and 72 h post-HS insult. Our findings expand on the knowledge about the time course of HS-induced oxidative damage and demonstrate for the first time that a single NAP injection dose-dependently prevents HS-induced oxidative damage to DNA and lipid membranes, in correlation with modulation of the glutathione system. Hence, NAP may represent a promising therapeutic strategy for avoiding HS-induced oxidative damage.

Potencial terapêutico das células-tronco de medula óssea no tratamento da epilepsia / Therapeutic potential of bone marrow stem cells in epilepsy treatment

Epilepsia é uma patologia bastante prevalente no nosso meio. Há um número significativo de pacientes que não obtém resposta com a terapêutica medicamentosa, o que motivou a pesquisa de novas terapêuticas. O conceito de neurogênese no cérebro adulto, hoje já amplamente conhecida, principalmente a que ocorre na zona subventricular e na zona subgranular do giro denteado, motivou o desenvolvimeto de técnicas que aproveitassem esse mecanismo na tentativa de obtenção de efeitos antiepileptogênicos e reparadores. A maior parte dos estudos em vigência hoje, e que buscam tal finalidade, trabalha o uso de transplante de células progenitoras neurais ou de células fetais em modelos experimentais. No entanto, a terapêutica com células-tronco de medula óssea parece bastante interessante e promissora. Em doenças neurológicas nas quais os danos são frequentemente irreversíveis, as estratégias regenerativas podem representar um novo caminho. Em nosso laboratório, temos estudado o potencial terapêutico de células-tronco da medula óssea no controle de crises espontâneas recorrentes associadas ao modelo da pilocarpina com resultados excelentes. Também já está em andamento o primeiro estudo em humanos utilizando células-tronco de medula óssea para o tratatamento da epilepsia.

Epilepsy is a prevalent pathology. A significative number of patients do not achieve an adequate response to pharmacological therapy. This observation has motivated the search for new strategies. The now well known concept of neurogenesis in the adult brain, in particular in the subventricular and dentate gyrus subgranular zones, has encouraged the development of strategies that might control this mechanism in order to obtain either antiepileptogenic or repair effects. Most studies focus on the transplantation of neural progenitor or fetal cells in experimental models, although bone marrow stem cell therapy is promising. In neurological diseases in which damage is frequently irreversible, regenerative strategies could represent a new path towards better treatment options. In our laboratory, we have been studying the therapeutic potential of bone marrow stem cells in controlling recurrent spontaneous seizures associated to the pilocarpine model of epilepsy with excellent results. We are also running the first study using bone marrow stem cell transplantation in the treatment of epilepsy in humans.