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Thirteen female Wistar rats were divided into two groups: one treated with ethanol and the other of untreated. Four newborns from each mother were selected and weighed, measured, and evaluated for physical characteristics. From these neonates, chondrocytes were extracted from the articular cartilages of the femur and tibia, and cultivated in a chondrogenic medium at 37oC and 5% CO2. At 7, 14, and 21 days of cultivation, alkaline phosphatase activity tests, MTT conversion to formazan, and percentage area covered by cells per field were performed. At 21 days, the percentage of PAS+ areas in 3D cultures was performed, as well as the evaluation of gene transcript expression for aggrecan, SOX-9, collagen type II, collagen X, Runx-2, and VEGF by real-time RT-PCR. The means were compared by Student's t-test. The weight of the ethanol group neonates was significantly lower than that of the controls. Chondrocyte cultures from the ethanol group showed significantly higher AP activity, MTT conversion, and cell percentage. There was higher expression of collagen type II and lower expression of SOX-9 in the ethanol group. There was no difference in the percentage of PAS+ areas in pellets and in expression of aggrecan, collagen X, Runx-2, or VEGF between groups. In conclusion, prenatal exposure to ethanol alters the phenotype and activity of offspring chondrocytes, which may be mechanisms by which endochondral bone formation is compromised by maternal ethanol consumption.

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Objetivou-se avaliar o efeito do dantrolene (DAN) e das células-tronco mesenquimais (CTM) no trauma espinhal agudo (TEA). Sessenta ratos Wistar foram divididos nos grupos CTM, DAN + CTM, DAN, trauma e placebo (TP) e sem trauma e placebo (STP). Realizou-se laminectomia de T12 em todos os grupos, seguida de TEA contusivo ∕ compressivo, com exceção do grupo STP. Uma hora depois, os grupos DAN + CTM e DAN receberam 10mg/kg de DAN. Após sete dias os grupos CTM e DAN + CTM receberam 1x106 células, por via intravenosa. Testes comportamentais foram realizados para avaliar a recuperação funcional durante 28 dias. Os animais traumatizados apresentaram paraplegia. Houve melhora funcional significativa nos grupos tratados com CTM, DAN ou associação DAN + CTM em comparação ao grupo TP (p<0,05). Conclui-se que o DAN e as CTM para tratamento de TEA em ratos apresentam efeitos neuroprotetores e promovem melhora neurológica funcional.(AU)

This study aimed to evaluate the effects of dantrolene (DAN) and mesenchymal stem cells (MSCs) in acute spinal cord injury (SCI). Sixty Wistar rats were divided into groups MSCs, MSCs + DAN, DAN, trauma and placebo (TP) and no trauma and placebo (STP). Laminectomy was performed at T12 level in all animals, followed by a weight-drop model of SCI, except for the STP group. An hour later, the MSCs + DAN and DAN groups received 10mg/kg of DAN. After seven days, the MSCs and MSCs + DAN groups received 1x106 cells intravenously. Behavioral tests were performed to assess functional recovery for 28 days. Traumatized animals showed paraplegia. There was a significant improvement in groups MSCs, DAN and MSCs + DAN compared to TP (p<0.05). It was concluded that DAN and MSCs for the treatment of SCI in rats have neuroprotection effect and promote functional neurological improvement.(AU)

Assuntos

RESUMO

Objetivou-se avaliar o efeito do dantrolene (DAN) e das células-tronco mesenquimais (CTM) no trauma espinhal agudo (TEA). Sessenta ratos Wistar foram divididos nos grupos CTM, DAN + CTM, DAN, trauma e placebo (TP) e sem trauma e placebo (STP). Realizou-se laminectomia de T12 em todos os grupos, seguida de TEA contusivo ∕ compressivo, com exceção do grupo STP. Uma hora depois, os grupos DAN + CTM e DAN receberam 10mg/kg de DAN. Após sete dias os grupos CTM e DAN + CTM receberam 1x106 células, por via intravenosa. Testes comportamentais foram realizados para avaliar a recuperação funcional durante 28 dias. Os animais traumatizados apresentaram paraplegia. Houve melhora funcional significativa nos grupos tratados com CTM, DAN ou associação DAN + CTM em comparação ao grupo TP (p<0,05). Conclui-se que o DAN e as CTM para tratamento de TEA em ratos apresentam efeitos neuroprotetores e promovem melhora neurológica funcional.(AU)

This study aimed to evaluate the effects of dantrolene (DAN) and mesenchymal stem cells (MSCs) in acute spinal cord injury (SCI). Sixty Wistar rats were divided into groups MSCs, MSCs + DAN, DAN, trauma and placebo (TP) and no trauma and placebo (STP). Laminectomy was performed at T12 level in all animals, followed by a weight-drop model of SCI, except for the STP group. An hour later, the MSCs + DAN and DAN groups received 10mg/kg of DAN. After seven days, the MSCs and MSCs + DAN groups received 1x106 cells intravenously. Behavioral tests were performed to assess functional recovery for 28 days. Traumatized animals showed paraplegia. There was a significant improvement in groups MSCs, DAN and MSCs + DAN compared to TP (p<0.05). It was concluded that DAN and MSCs for the treatment of SCI in rats have neuroprotection effect and promote functional neurological improvement.(AU)

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RESUMO

ABSTRACT: This study aimed to evaluate the effects of dantrolene (DAN) and mesenchymal stem cells (MSCs) in acute spinal cord injury (SCI). Sixty Wistar rats were divided into groups MSCs, MSCs + DAN, DAN, trauma and placebo (TP) and no trauma and placebo (STP). Laminectomy was performed at T12 level in all animals, followed by a weight-drop model of SCI, except for the STP group. An hour later, the MSCs + DAN and DAN groups received 10mg/kg of DAN. After seven days, the MSCs and MSCs + DAN groups received 1x106 cells intravenously. Behavioral tests were performed to assess functional recovery for 28 days. Traumatized animals showed paraplegia. There was a significant improvement in groups MSCs, DAN and MSCs + DAN compared to TP (p 0.05). It was concluded that DAN and MSCs for the treatment of SCI in rats have neuroprotection effect and promote functional neurological improvement.

RESUMO: Objetivou-se avaliar o efeito do dantrolene (DAN) e das células-tronco mesenquimais (CTM) no trauma espinhal agudo (TEA). Sessenta ratos Wistar foram divididos nos grupos CTM, DAN + CTM, DAN, trauma e placebo (TP) e sem trauma e placebo (STP). Realizou-se laminectomia de T12 em todos os grupos, seguida de TEA contusivo compressivo, com exceção do grupo STP. Uma hora depois, os grupos DAN + CTM e DAN receberam 10mg/kg de DAN. Após sete dias os grupos CTM e DAN + CTM receberam 1x106 células, por via intravenosa. Testes comportamentais foram realizados para avaliar a recuperação funcional durante 28 dias. Os animais traumatizados apresentaram paraplegia. Houve melhora funcional significativa nos grupos tratados com CTM, DAN ou associação DAN + CTM em comparação ao grupo TP (p 0,05). Conclui-se que o DAN e as CTM para tratamento de TEA em ratos apresentam efeitos neuroprotetores e promovem melhora neurológica funcional.

RESUMO

This study aimed to evaluate the effect of methylprednisolone sodium succinate, dantrolene sodium, and their combination on experimental spinal cord injury. We used 25 rats (Rattus norvegicus) that were divided into five groups. The negative control group (NC) consisted of animals without spinal cord trauma. In the groups with spinal cord trauma, the positive control group (PC) was given no treatment, the MS group was treated with methylprednisolone, the MS/DS group was treated with methylprednisolone and dantrolene, and the DS group was treated with dantrolene alone. The animals' motor function was evaluated daily, as measured with the open field test. Eight days after surgery, the animals were euthanized for spinal cord collection. Descriptive morphological evaluation, anti-NeuN immunohistochemistry, TUNEL, and anti-Bax immunofluorescence were performed. There was no significant difference between the PC, MS, MS/DS and DS groups with respect to BBB scores, neuronal and glial staining, or Bax expression (P < 0.05). Therefore, we conclude that methylprednisolone sodium succinate, dantrolene sodium, or the combination of these drugs did not reduce neuronal and glial loss, intrinsic pathway apoptosis, or promote functional recovery.

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Excessive accumulation of intracellular calcium is the most critical step after spinal cord injury (SCI). Reducing the calcium influx should result in a better recovery from SCI. Calcium channel blockers have been shown a great potential in reducing brain and spinal cord injury. In this study, we first tested the neuroprotective effect of MVIIC on slices of spinal cord subjected to ischemia evaluating cell death and caspase-3 activation. Thereafter, we evaluated the efficacy of MVIIC in ameliorating damage following SCI in rats, for the first time in vivo. The spinal cord slices subjected a pretreatment with MVIIC showed a cell protection with a reduction of dead cells in 24.34% and of caspase-3-specific protease activation. In the in vivo experiment, Wistar rats were subjected to extradural compression of the spinal cord at the T12 vertebral level using a weigh of 70 g/cm, following intralesional treatment with either placebo or MVIIC in different doses (15, 30 and 60 pmol) five minutes after injury. Behavioral testing of hindlimb function was done using the Basso Beattie Bresnahan locomotor rating scale, and revealed significant recovery with 15 pmol (G15) compared to other trauma groups. Also, histological bladder structural revealed significant outcome in G15, with no morphological alterations, and anti-NeuN and TUNEL staining showed that G15 provided neuron preservation and indicated that this group had fewer neuron cell death, similar to sham. These results showed the neuroprotective effects of MVIIC in in vitro and in vivo model of SCI with neuronal integrity, bladder and behavioral improvements.

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BACKGROUND: Calcium channel blockers such as conotoxins have shown a great potential to reduce brain and spinal cord injury. MVIIC neuroprotective effects analyzed in in vitro models of brain and spinal cord ischemia suggest a potential role of this toxin in preventing injury after spinal cord trauma. However, previous clinical studies with MVIIC demonstrated that clinical side effects might limit the usefulness of this drug and there is no research on its systemic effects. Therefore, the present study aimed to investigate the potential toxic effects of MVIIC on organs and to evaluate clinical and blood profiles of rats submitted to spinal cord injury and treated with this marine toxin. Rats were treated with placebo or MVIIC (at doses of 15, 30, 60 or 120 pmol) intralesionally following spinal cord injury. Seven days after the toxin administration, kidney, brain, lung, heart, liver, adrenal, muscles, pancreas, spleen, stomach, and intestine were histopathologically investigated. In addition, blood samples collected from the rats were tested for any hematologic or biochemical changes. RESULTS: The clinical, hematologic and biochemical evaluation revealed no significant abnormalities in all groups, even in high doses. There was no significant alteration in organs, except for degenerative changes in kidneys at a dose of 120 pmol. CONCLUSIONS: These findings suggest that MVIIC at 15, 30 and 60 pmol are safe for intralesional administration after spinal cord injury and could be further investigated in relation to its neuroprotective effects. However, 120 pmol doses of MVIIC may provoke adverse effects on kidney tissue.

RESUMO

Calcium channel blockers such as conotoxins have shown a great potential to reduce brain and spinal cord injury. MVIIC neuroprotective effects analyzed in in vitro models of brain and spinal cord ischemia suggest a potential role of this toxin in preventing injury after spinal cord trauma. However, previous clinical studies with MVIIC demonstrated that clinical side effects might limit the usefulness of this drug and there is no research on its systemic effects. Therefore, the present study aimed to investigate the potential toxic effects of MVIIC on organs and to evaluate clinical and blood profiles of rats submitted to spinal cord injury and treated with this marine toxin. Rats were treated with placebo or MVIIC (at doses of 15, 30, 60 or 120 pmol) intralesionally following spinal cord injury. Seven days after the toxin administration, kidney, brain, lung, heart, liver, adrenal, muscles, pancreas, spleen, stomach, and intestine were histopathologically investigated. In addition, blood samples collected from the rats were tested for any hematologic or biochemical changes.

Assuntos

RESUMO

Calcium channel blockers such as conotoxins have shown a great potential to reduce brain and spinal cord injury. MVIIC neuroprotective effects analyzed in in vitro models of brain and spinal cord ischemia suggest a potential role of this toxin in preventing injury after spinal cord trauma. However, previous clinical studies with MVIIC demonstrated that clinical side effects might limit the usefulness of this drug and there is no research on its systemic effects. Therefore, the present study aimed to investigate the potential toxic effects of MVIIC on organs and to evaluate clinical and blood profiles of rats submitted to spinal cord injury and treated with this marine toxin. Rats were treated with placebo or MVIIC (at doses of 15, 30, 60 or 120 pmol) intralesionally following spinal cord injury. Seven days after the toxin administration, kidney, brain, lung, heart, liver, adrenal, muscles, pancreas, spleen, stomach, and intestine were histopathologically investigated. In addition, blood samples collected from the rats were tested for any hematologic or biochemical changes.

Assuntos

RESUMO

Calcium channel blockers such as conotoxins have shown a great potential to reduce brain and spinal cord injury. MVIIC neuroprotective effects analyzed in in vitro models of brain and spinal cord ischemia suggest a potential role of this toxin in preventing injury after spinal cord trauma. However, previous clinical studies with MVIIC demonstrated that clinical side effects might limit the usefulness of this drug and there is no research on its systemic effects. Therefore, the present study aimed to investigate the potential toxic effects of MVIIC on organs and to evaluate clinical and blood profiles of rats submitted to spinal cord injury and treated with this marine toxin. Rats were treated with placebo or MVIIC (at doses of 15, 30, 60 or 120 pmol) intralesionally following spinal cord injury. Seven days after the toxin administration, kidney, brain, lung, heart, liver, adrenal, muscles, pancreas, spleen, stomach, and intestine were histopathologically investigated. In addition, blood samples collected from the rats were tested for any hematologic or biochemical changes.(AU)

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