Léder Leal Xavier

Endurance and Resistance Exercise Training Programs Elicit Specific Effects on Sciatic Nerve Regeneration After Experimental Traumatic Lesion in Rats

Objective. To evaluate the effects of endurance, resistance, and a combination of both types of exercise training on hindlimb motor function recovery and nerve regeneration after experimental sciatic nerve lesion in rats. Methods. Sciatic nerve crush was performed on adult male rats, and after 2 weeks of the nerve lesion, the animals were submitted to endurance, resistance, and a combination of endurance-resistance training programs for 5 weeks. Over the training period, functional recovery was monitored weekly using the Sciatic Functional Index (SFI) and histological and morphometric nerve analyses were used to assess the nerve regeneration at the end of the trainings. Results. The SFI values of the endurance-trained group reached the control values from the first posttraining week and were significantly better than both the resistance-trained group at the first, second, and third posttraining weeks and the concurrent training group at the first posttraining week. At the distal portion of the regenerating sciatic nerve, the endurance-trained group showed a greater degree of the myelinated fiber maturation than the sedentary, resistance-trained, and concurrent training groups. Furthermore, the endurance-trained group showed a smaller percentage area of endoneurial connective tissue and a greater percentage area of myelinated fibers than the sedentary group. Conclusion . These data provide evidence that endurance training improves sciatic nerve regeneration after an experimental traumatic injury and that resistance training or the combination of 2 strategies may delay functional recovery and do not alter sciatic nerve fiber regeneration.

Hemispheric brain injury and behavioral deficits induced by severe neonatal hypoxia-ischemia in rats are not attenuated by intravenous administration of human umbilical cord blood cells.

Neonatal hypoxia-ischemia (HI) is an important cause of mortality and morbidity in infants. Human umbilical cord blood (HUCB) is a potential source of cellular therapy in perinatology. We investigated the effects of HUCB cells on spatial memory, motor performance, and brain morphologic changes in neonate rats submitted to HI. Seven-day-old rats underwent right carotid artery occlusion followed by exposure to 8% O2 inhalation for 2 h. Twenty-four hours after HI, rats received either saline solution or HUCB cells i.v. After 3 wk, rats were assessed using a Morris Water Maze and four motor tests. Subsequently, rats were killed for histologic, immunohistochemical, and polymerase chain reaction (PCR) analyses. HI rats showed significant spatial memory deficits and a volumetric decrease in the hemisphere ipsilateral to arterial occlusion. These deficits and decreases were not significantly attenuated by the injection of HUCB cells. Moreover, immunofluorescence and PCR analysis revealed few HUCB cells located in rat brain. Intravenous administration of HUCB cells requires optimization to achieve improved therapeutic outcomes in neonatal hypoxic-ischemic injury.

Morphological changes in hippocampal astrocytes induced by environmental enrichment in mice.

Environmental enrichment is known to induce plastic changes in the brain, including morphological changes in hippocampal neurons, with increases in synaptic and spine densities. In recent years, the evidence for a role of astrocytes in regulating synaptic transmission and plasticity has increased, and it is likely that morphological and functional changes in astrocytes play an important role in brain plasticity. Our study was designed to evaluate changes in astrocytes induced by environmental enrichment in the CA1 region of the hippocampus, focusing on astrocytic density and on morphological changes in astrocytic processes. After 8 weeks of environmental enrichment starting at weaning, male CF-1 mice presented no significant changes in astrocyte number or in the density of glial fibrillary acidic protein (GFAP) immunoreactivity in the stratum radiatum. However, they did present changes in astrocytic morphology in the same region, as expressed by a significant increase in the ramification of astrocytic processes measured by the Sholl concentric circles method, as well as by an increase in the number and length of primary processes extending in a parallel orientation to CA1 nerve fibers. This led astrocytes to acquire a more stellate morphology, a fact which could be related to the increase in hippocampal synaptic density observed in previous studies. These findings corroborate the idea that structural changes in astrocytic networks are an integral part of plasticity processes occurring in the brain.

Evaluation of chronic omega-3 fatty acids supplementation on behavioral and neurochemical alterations in 6-hydroxydopamine-lesion model of Parkinson's disease

Omega-3 polyunsaturated fatty acids (omega-3 PUFAs) have been widely associated to beneficial effects over different neuropathologies, but only a few studies associate them to Parkinsons disease (PD). Rats were submitted to chronic supplementation (21-90 days of life) with fish oil, rich in omega-3 PUFAs, and were uni- or bilaterally lesioned with 4microg of the neurotoxin 6-hydroxydopamine (6-OHDA) in the medial forebrain bundle. Although lipid incorporation was evidenced in neuronal membranes, it was not sufficient to compensate motor deficits induced by 6-OHDA. In contrast, omega-3 PUFAs were capable of reducing rotational behavior induced by apomorphine, suggesting neuroprotection over dyskinesia. The beneficial effects of omega-3 PUFAs were also evident in the maintenance of thiobarbituric acid reactive substances index from animals lesioned with 6-OHDA similar to levels from SHAM and intact animals. Although omega-3 PUFAs did not modify the tyrosine hydroxylase immunoreactivity in the substantia nigra pars compacta and in the ventral tegmental area, nor the depletion of dopamine (DA) and its metabolites in the striatum, DA turnover was increased after omega-3 PUFAs chronic supplementation. Therefore, it is proposed that omega-3 PUFAs action characterizes the adaptation of remaining neurons activity, altering striatal DA turnover without modifying the estimated neuronal population.

Plastic changes induced by neonatal handling in the hypothalamus of female rats.

Early-life events can exert profound long-lasting effects on several behaviors such as fear/anxiety, sexual activity, stress responses and reproductive functions. Present study aimed to examine the effects of neonatal handling on the volume and number of cells in the hypothalamic paraventricular nucleus (pPVN, parvocellular and mPVN, magnocellular regions) and the supraoptic nucleus (SON) in female rats at 11 and 90 days of age. Moreover, in the same areas, immunohistochemistry for oxytocin (OT) and glial fibrillary acidic protein (GFAP) were analyzed in the adult animals. Daily handling during the first 10 postnatal days reduced the number of cells in the pPVN and SON at both the 11 and 90 days. Handling decreased the number of OT-positive parvocellular cells in the PVN in adult females. No significant differences were detected on the optical density (OD) of GFAP-positive cells between the handled and nonhandled adult females. The effect of handling on cell loss was observed 24 h after the 10-day handling period and persisted into adulthood, indicating a stable morphological trace. Results suggest that neonatal handling can induce plastic changes in the central nervous system.

Influence of sex and estrous cycle, but not laterality, on the neuronal somatic volume of the posterodorsal medial amygdala of rats.

The aim of the present study was to measure the cell body volume of neurons from the posterodorsal subnucleus of the medial amygdala (MePD) of adult male (n=5) and diestrus, proestrus and estrus female (n=4-5 in each group) rats to reveal a possible sexual dimorphism, estrous cycle variations and laterality in this morphological parameter. The brains of adult Wistar rats were sectioned (1 microm), stained with 1% toluidine blue and the stereological estimation of neuronal soma volume of both sides of MePD was realized using the Cavalieri method and the technique of point counting. Data were compared by a two-way ANOVA for repeated measures and the least significance difference post hoc test. In the MePD, mean neuronal somatic volume showed a statistical difference among groups (p=0.005), but neither an effect of laterality (p=0.33) nor interactions between groups and laterality (p=0.78) were found. Post hoc test showed that males (mean+/-S.E.M., 2075.67+/-135.79 microm(3)) have larger mean neuronal somatic volume compared to females in proestrus (1503.30+/-44.46 microm(3)) and in estrus (1616.69+/-71.49 microm(3), p<0.05 in both cases), but not in diestrus (1940.78+/-129.68 microm(3), p>0.05). Moreover, diestrus females displayed larger mean neuronal somatic volume than proestrus female rats (p<0.05). It is suggested that neuronal somatic volume is another sexually dimorphic finding in the MePD, for which it is relevant to set apart the different phases of the estrous cycle to reveal the presence of gonadal hormones effects in the rat MePD neurons.

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 Racines 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.

Respiratory muscle training improves hemodynamics, autonomic function, baroreceptor sensitivity, and respiratory mechanics in rats with heart failure

Respiratory muscle training (RMT) improves functional capacity in chronic heart-failure (HF) patients, but the basis for this improvement remains unclear. We evaluate the effects of RMT on the hemodynamic and autonomic function, arterial baroreflex sensitivity (BRS), and respiratory mechanics in rats with HF. Rats were assigned to one of four groups: sedentary sham (n = 8), trained sham (n = 8), sedentary HF (n = 8), or trained HF (n = 8). Trained animals underwent a RMT protocol (30 min/day, 5 day/wk, 6 wk of breathing through a resistor), whereas sedentary animals did not. In HF rats, RMT had significant effects on several parameters. It reduced left ventricular (LV) end-diastolic pressure (P < 0.01), increased LV systolic pressure (P < 0.01), and reduced right ventricular hypertrophy (P < 0.01) and pulmonary (P < 0.001) and hepatic (P < 0.001) congestion. It also decreased resting heart rate (HR; P < 0.05), indicating a decrease in the sympathetic and an increase in the vagal modulation of HR. There was also an increase in baroreflex gain (P < 0.05). The respiratory system resistance was reduced (P < 0.001), which was associated with the reduction in tissue resistance after RMT (P < 0.01). The respiratory system and tissue elastance (Est) were also reduced by RMT (P < 0.01 and P < 0.05, respectively). Additionally, the quasistatic Est was reduced after RMT (P < 0.01). These findings show that a 6-wk RMT protocol in HF rats promotes an improvement in hemodynamic function, sympathetic and vagal heart modulation, arterial BRS, and respiratory mechanics, all of which are benefits associated with improvements in cardiopulmonary interaction.

Gonadal hormone regulation of glial fibrillary acidic protein immunoreactivity in the medial amygdala subnuclei across the estrous cycle and in castrated and treated female rats

The medial amygdala (MeA) is a sexually dimorphic area that modulates neuroendocrine and behavioral activities and where gonadal hormones play an important role in neuron-glial and synaptic plasticity. Immunohistochemistry was used to identify the astrocytic marker glial fibrillary acidic protein (GFAP) in the different MeA subnuclei--anterodorsal (MeAD), posterodorsal (MePD) and posteroventral (MePV)--of intact female rats in the different phases of the estrous cycle and in ovariectomized females treated with hormonal substitutive therapy. Data semi-quantified by optical densitometry showed that, in the proestrus phase, the GFAP immunoreactivity (GFAP-ir) was higher when compared to the other phases of the estrous cycle (P < 0.02). GFAP-ir was also higher in the MePD than in the MeAD or in the MePV (P < 0. 02). In ovariectomized females, injections of estradiol alone or estradiol plus progesterone increased GFAP-ir in the MePD and in the MePV (P < 0.001), but not in the MeAD (P > 0.3), when compared to control data. These findings suggest that astrocytic GFAP in the MeA subnuclei can be affected either by physiological levels or by hormonal manipulation of the ovarian steroids, which may contribute to the plasticity of local and integrated functional activities of these brain areas in female rats.

New Therapy of Skin Repair Combining Adipose-Derived Mesenchymal Stem Cells with Sodium Carboxymethylcellulose Scaffold in a Pre-Clinical Rat Model

Lesions with great loss of skin and extensive burns are usually treated with heterologous skin grafts, which may lead rejection. Cell therapy with mesenchymal stem cells is arising as a new proposal to accelerate the healing process. We tested a new therapy consisting of sodium carboxymethylcellulose (CMC) as a biomaterial, in combination with adipose-derived stem cells (ADSCs), to treat skin lesions in an in vivo rat model. This biomaterial did not affect membrane viability and induced a small and transient genotoxicity, only at the highest concentration tested (40 mg/mL). In a rat wound model, CMC at 10 mg/mL associated with ADSCs increased the rate of cell proliferation of the granulation tissue and epithelium thickness when compared to untreated lesions (Sham), but did not increase collagen fibers nor alter the overall speed of wound closure. Taken together, the results show that the CMC is capable to allow the growth of ADSCs and is safe for this biological application up to the concentration of 20 mg/mL. These findings suggest that CMC is a promising biomaterial to be used in cell therapy.