Sandra Gomes Valente

Effects of pinealectomy and the treatment with melatonin on the temporal lobe epilepsy in rats

The aim of the present work was to analyze the effects of pinealectomy in the development of the epilepsy model induced by pilocarpine in adult male rats. Group I: Wistar male adult rats were submitted to pinealectomy, and 7 days after surgery, these animals received pilocarpine (350 mg/kg, i.p.) to induce three distinct behavioral phases: status epilepticus, seizure-free, and chronic phases. This late, as well as all control groups were continuously video-recorded for 60 days, to study behavior parameters. These animals were killed and the brain sections were processed for Nissl and neo-Timm. Group II: Another group, also submitted to pinealectomy, received several injections of melatonin (2.5 mg/kg): 20 min before, concomitantly with pilocarpine, 30 min, 1 h, and 2 h after pilocarpine administration. Some animals from group I and all from group II were sacrificed 48 h following status epilepticus onset to perform TUNEL assay. The latency for status epilepticus onset, status epilepticus length as well as mortality rate during status epilepticus were similar for pinealectomized and control groups. On the other hand, pinealectomized rats presented minor duration of the silent period, a higher number of spontaneous seizures during the chronic phase, increased number of TUNEL-positive cells (acute phase), increased neuronal loss, and marked supragranullar mossy fibers sprouting (chronic phase) in the hippocampal formation, when compared with control groups. Our data show that the pinealectomy facilitates the epileptogenic process that follows the long-lasting status epilepticus. This facilitation can be partially reverted by the simultaneous administration of melatonin.

Bone marrow cells are able to increase vessels number during repair of sciatic nerve lesion

The aim of this study was to compare the outcomes of nerve autografts (GRF) and venous grafts containing mononuclear bone marrow cells (BMCs) in sciatic nerve-lesioned rats. Control animals underwent sham operations (SHAM), received empty venous grafts (EPV), or received venous grafts containing BMC vehicle (AGR). Outcome was evaluated through sciatic functional index (SFI), morphometric and morphologic analyses of the nerve distal to the lesion, and the number of spinal cord motor neurons positive for the retrograde tracer, Fluoro-Gold. All groups exhibited poor results in SFI when compared to SHAM animals throughout the postoperative period. All groups also had a significantly greater fiber density, decreased fiber diameter, and decreased motor neuron number than the SHAM group. No significant difference between the GRF and BMC groups was observed in any of these parameters. On the other hand, vessel density was significantly higher in BMC than all other groups. BMC-containing venous grafts are superior to nerve autografts in increasing vessel density during sciatic nerve regeneration.

Castration in female rats modifies the development of the pilocarpine model of epilepsy

Previous studies have shown that the susceptibility to pilocarpine-induced status epilepticus (SE) in female rats changes according to estrous cycle phases. These studies have also shown that following pilocarpine administration changes occur in gonadal, hypophyseal and hypothalamic hormones that could contribute for the sequence of the epileptic events. Accordingly, the present work aimed to investigate the role of sexual hormones withdrawal on the development of the pilocarpine model of epilepsy in female rats. With this purpose, castrated and non-castrated adult female Wistar rats were injected with pilocarpine and some characteristic parameters of the experimental model were observed. The results showed increased mortality after pilocarpine injection in the castrated rats when compared with non-castrated females. The latency period for SE onset and for the first spontaneous seizure was decreased in castrated when compared with non-castrated animals. The mossy fiber sprouting measured by neo-Timm scale during the chronic period, reached grade 3 for castrated epileptic rats while the non-castrated epileptic rats showed grade 2. Our results indicate that castration interferes with the epileptogenesis in the pilocarpine model of epilepsy suggesting that female sexual hormones could have protective effects against pilocarpine-induced SE.

Estudo da regeneração nervosa em nervos tibiais de ratos wistar utilizando o Fluoro-Gold® como marcador neuronal

The authors studied, in rats, of nerve regeneration in nerve grafts comparing with control group using Fluoro-Gold® (FG) labeled motor neurons count in spinal cord. In control group both tibial nerves were exposed to FG® and motor neurons counted in spinal cord in 48 hours, after perfusion, in a medular segment from L3 to S1. In experimental group, ressecting 8 mm, a gap was created in both tibial nerves and nerve segment of one nerve was used to repair contralateral side in a traditional nerve graft suture. After four months, right tibial nerve was exposed to FG® distal to graft and left tibial nerve exposed to FG proximal to graft. In 48 hours, after perfusion, motoneurons were counted in a medular segment from L3 to S1. In both groups medular segment was cut in 40 mm slices and all labeled cells counted. Wilcoxon and Student tests were used for statistical analysis. Control group presented a significative increased number of motoneurons when compared to experimental group. In experimental group number of motoneurons was significantly decreased when tibial nerve was exposed to FG® distal to nerve graft. Nerve graft promoted a partial block to axon migration after four months of surgery.

Nerve Regeneration: Is There an Alternative to Nervous Graft?

BACKGROUND In nerve injury with nervous gap, no restitution method was found better than the autograft, however, it has the disadvantage of damaging a normal nerve to be used as a graft. Platelet-rich plasma (PRP) is a possible filler material for vein grafts used as conduits for nerve regeneration, preventing its collapse, and providing growth factors and osteoconductive proteins. METHODS Isogenic rats were randomly divided into three groups. They received nerve autografts (GRF), PRP-containing vein grafts or a sham operation. Outcomes were evaluated by the sciatic functional index (SFI), morphometric, and morphologic analyses of the nerve distal to the lesion, and the number of spinal cord motoneurons positive for retrograde Fluoro-Gold (Santa Cruz Biotechnology, Inc., Dallas, TX) tracer. RESULTS The PRP and GRF groups had lower SFI values than the control animals throughout the postoperative period. The SFI was significantly higher in the PRP group than the GRF group at 90 days postoperatively (p = 0.011). Fiber diameter and number of motoneurons were significantly decreased in both the PRP and GRF groups, as compared with the control. CONCLUSION PRP within a vein conduit may be an effective alternative or adjuvant to GRF, the current preferred treatment for nerve injury with a nerve gap, and further investigations are required to fully define the role of PRP in nerve regeneration.

Estudo da regeneração de nervos tibiais de ratos Wistar em sutura primária com "gap" e sem "gap", cobertos por segmentos de veia

Objective This study compared nerve regeneration in Wistar rats, using epineural neurorrhaphy with a gap of 1.0 mm and without a gap, both wrapped with jugular vein tubes. Motor neurons in the spinal cord between L3 and S1 were used for the count, marked by exposure of the tibial nerve to Fluoro-Gold (FG). Method The tibial nerves on both sides were cut and sutured, with a gap on one side and no gap in the other. The sutures were wrapped with a jugular vein. Four months after surgery the tibial nerves were exposed to Fluoro-Gold and the motor neuron count performed in the spinal cord. Results The results were statistically analyzed by the paired Wilcoxon test. There was a statistical difference between the groups with and without gap in relation to the motor neuron count (p=0.013). Conclusion The epineural neurorraphy without gap wrapped with jugular vein showed better results for nerve regeneration than the same procedure with gap. Level of Evidence: Experimental Study.OBJECTIVE: This study compared nerve regeneration in Wistar rats, using epineural neurorrhaphy with a gap of 1.0 mm and without a gap, both wrapped with jugular vein tubes. Motor neurons in the spinal cord between L3 and S1 were used for the count, marked by exposure of the tibial nerve to Fluoro-Gold (FG). METHOD: The tibial nerves on both sides were cut and sutured, with a gap on one side and no gap in the other. The sutures were wrapped with a jugular vein. Four months after surgery the tibial nerves were exposed to Fluoro-Gold and the motor neuron count performed in the spinal cord. RESULTS: The results were statistically analyzed by the paired Wilcoxon test. There was a statistical difference between the groups with and without gap in relation to the motor neuron count (p=0.013). CONCLUSION: The epineural neurorraphy without gap wrapped with jugular vein showed better results for nerve regeneration than the same procedure with gap. Level of Evidence: Experimental Study.

Effect of aerobic physical exercise in pinealectomized animals submitted to the pilocarpine model of epilepsy

OBJECTIVE: To better clarify the positive effects of physical exercise in the epilepsy, we analyzed the effect of the pinealectomy in animals with temporal lobe epilepsy (TLE) induced by pilocarpine submitted to an aerobic physical program. MATERIAL AND METHODS: Forty adults Wistar rats were used: 1) PX + CHRONIC - Pinealectomized animals (PX) with TLE (CHRONIC) without exercise (n = 9); 2) PX + CHRONIC + EXERCISE - submitted to an aerobic physical exercise program (n = 5); 3) CHRONIC - without exercise (n = 8); 4) CHRONIC + EXERCISE (n = 8); 5) CTRL - control without exercise (n = 5); 6) CTRL + EXERCISE (n = 5). The physical exercise program consisted of 1 hour of treadmill, 5 days/week, during 30 days, at 60% VO2max. The Nissl and neo-Timm methods were used. RESULTS: The pinealectomy increased the frequency of seizures in animals with epilepsy. It was observed a reduction of the neuronal death and mossy fiber sprouting in the animals with epilepsy submitted to an aerobic physical exercise program. However, the physical exercise program did not modify the frequency of the seizures in the pinealectomized animals.