Benefícios da condroitinase abc associada a células-tronco mesenquimais na lesão espinhal aguda em ratos / Benefits of chondroitinase abc associated with mesenchymal stem cells in acute spinal cord injury in rats

Com o objetivo de estudar o efeito da condroitinase associada às células-tronco mesenquimais na lesão aguda da medula espinhal, utilizaram-se 50 ratos Lewis, distribuídos igualmente nos grupos: controle negativo (CN), tratamento com placebo (PLA), condroitinase (CDN), células-tronco mesenquimais (CTM) e condroitinase mais células-tronco mesenquimais (CDN+CTM). Todos os animais tiveram a medula espinhal exposta por laminectomia, e os grupos PLA, CDT, CTM e CDT+CTM sofreram também trauma medular compressivo. Após sete dias, procedeu-se à reexposição da medula espinhal, quando os grupos PLA e CTM receberam 4µL de líquido cefalorraquidiano artificial via intralesional, e os grupos CDT e CDT+CTM receberam o mesmo líquido contendo 2,2U de condroitinase. Após 14 dias da cirurgia inicial, todos os animais receberam 0,2mL de PBS via endovenosa, contudo, nos grupos CTM e CDT+CTM, esse líquido continha 1x106 CTM. Avaliou-se a capacidade motora até o 28o dia pós-trauma e, posteriormente, as medulas espinhais foram analisadas por RT-PCR, para quantificação da expressão gênica para BDNF, NT-3, VEGF, KDR e PECAM-1, e por imunoistoquímica, para detecção das células-tronco GFP injetadas (anti-GFP), quantificação dos neurônios (anti-NeuN) e da GFAP e vimentina, para avaliação da cicatriz glial. As análises estatísticas foram realizadas com o auxílio do Prism 5 for Windows, com o nível de significância de 5%. Não houve diferença entre os grupos quanto à capacidade motora. O grupo CDT+CTM apresentou maior imunoexpressão de neurônios viáveis do que o placebo. No CTM, houve maior expressão dos fatores neurotróficos BDNF e VEGF. E no CDT, houve menor imunoexpressão de vimentina. Concluiu-se que a associação CDT+CTM favorece a viabilidade neuronal após o trauma, que o tratamento com CTM promove aumento na expressão dos fatores tróficos BDNF e VEGF e que o tratamento com condroitinase é efetivo na redução da cicatriz glial.(AU)

The aim of this work was to study the effect of chondroitinase associated with mesenchymal stem cells in acute spinal cord injury. Therefore, 50 Lewis rats were distributed in the following groups: negative control (NC), treatment with placebo (PLA), chondroitinase (CDT), mesenchymal stem cells (MSC), and chondroitinase associated with mesenchymal stem cells (CDT + MSC). All animals had their spinal cord exposed by laminectomy, and the groups named PLA, CDT, MSC and CDT + MSC also suffered compressive spinal cord trauma. After seven days, the spinal cord was re-exposed, when the PLA and MSCs groups received 4uL of artificial cerebrospinal fluid through the lesion, and the CDT group and CDT + MSC received the same fluid containing 2,2U of chondroitinase. 14 days after the first surgery, all animals received 0.2ml of PBS intravenously; however, the MSC and CDT + MSC groups received the same liquid also containing 1x106 MSCs. The motor skills were evaluated up to 28 days post-injury and, subsequently, the spinal cords were analyzed by RT-PCR for BDNF, NT-3, VEGF, PECAM-1 and KDR gene expression quantification, immunohistochemistry to detect injected stem cells GFP (anti-GFP), to quantify neurons (anti-NeuN), GFAP and detect vimentin in order to evaluate the glial scar. Statistical analyzes were performed by Prism 5 for Windows using a 5% level of significance. There was no difference between groups with regarding motor capacity. The CDT + MSC group showed increased immunoreactivity of viable neurons than placebo. In MSC, there was a greater expression of neurotrophic factors BDNF and VEGF. Also, there was less vimentin immunostaining in group CDT. It was concluded that CDT + MSC association promotes neuronal viability after trauma, in which treatment with MSC promotes increased expression of BDNF and VEGF trophic factors, and also that treatment with chondroitinase is effective in reducing the glial scar.(AU)

Immunomodulatory and neuroprotective effect of cryopreserved allogeneic mesenchymal stem cells on spinal cord injury in rats.

This study aimed to evaluate the immunomodulatory and neuroprotective effects of allogeneic and cryopreserved mesenchymal stem cells (MSCs) on spinal cord injury. A total of 120 rats were distributed into the following groups: negative control (NC) - without injury, positive control (PC) - with injury without treatment, and group treated with MSC (GMSC) - with injury and treated. Motor function was evaluated by the BBB test at 24, 48, and 72 h and at 8 and 21 postoperative days. Spinal cords were evaluated by histopathology and immunohistochemistry to determine the expression of CD68, NeuN, and GFAP. IL-10, TNF-α, IL-1ß, TGF-ß, BDNF, GDNF, and VEGF expression was quantified by RT-PCR. The GMSC presented higher scores for motor function at 72 h and 8 and 21 days after injury, lower expression of CD68 at 8 days, and lower expression of GFAP at 21 days compared to the PC. In addition, higher expression of NeuN and lower degeneration of the white matter occurred at 21 days. The GMSC also showed higher expression of IL-10 24 h after injury, GDNF at 48 h and 8 days, and VEGF at 21 days. Moreover, lower expression of TNF-α was observed at 8 and 21 days and TGF-ß at 24 h and 21 days. There were no differences in the expression of IL-1ß and BDNF between the GMSC and PC. Thus, cryopreserved MSCs promote immunomodulatory and neuroprotective effects in rats with spinal cord injury by increasing IL-10, GDNF, and VEGF expression and reducing TNF-α and TGF-ß expression.

Osteointegração da hidroxiapatita sintética no processo alveolar da mandíbula de cães: aspectos clínicos e radiográficos / Osteointegration of synthetic hydroxyapatite in alveolar process of the jaw of dogs: clinical and radiographic aspects

Avaliou-se o efeito da hidroxiapatita sintética na regeneração do osso alveolar e o efeito no tecido vivo de cães. Em dois grupos de 14 cães adultos hígidos, pesando entre 10 e 15kg, foram criados defeitos de 6 x 5mm na superfície vestibular do processo alveolar mandibular direito até atingir a raiz do quarto pré-molar. Em um grupo (tratado), o defeito foi preenchido com hidroxiapatita sintética; o grupo sem tratamento foi usado como controle. Efetuaram-se avaliações clínicas diárias durante uma semana, avaliações radiográficas após a cirurgia e aos oito, 21, 42, 60, 90 e 120 dias do pós-operatório. Vinte e quatro cães apresentaram inflamação, sendo a recuperação no grupo tratado mais lenta. Todos os animais tiveram sangramento com a hidroxiapatita. No grupo-controle houve aumento crescente da radiopacidade dos defeitos, no entanto, aos 120 dias do pós-operatório, os defeitos ainda eram visíveis. No grupo tratado, inicialmente a radiopacidade foi maior que a do osso normal, com diminuição gradual até se tornar semelhante à do osso vizinho, 60 dias após a cirurgia. A hidroxiapatita T290800-1 acelerou o preenchimento do defeito provocado no processo alveolar e acarretou inflamação e hemorragia gengival, o que, no entanto, não contra-indicou o seu uso.

The effect of the synthetic hydroxyapatite on the regeneration of the alveolar bone of dogs and on the alive tissue were evaluated. In two groups of 14 adult healthy dogs, weighing from 10 to 15kg, some defects around 6 x 5mm were provoked on the vestibular surface of the right jaw alveolar process until reaching the root of the forth premolar. In one group, the defect was infilled with synthetic hydroxyapatite. The no treated group dogs were used as control. Clinical evaluations were daily performed over one week, as well as radiographic evaluations after surgery, and on 8, 21, 42, 60, 90 and 120 days after surgery. Twenty-four dogs showed inflammation and the recovery in the treated group was slower than in the control group. Bleeding at the presence of hydroxyapatite was observed in all the animals. The radiographic examination presented an increasing radiopacity in the control group; however, this defect was still visible on day 120 after operation. Initially, in the treated group, radiopacity was higher than that of the normal bone and a gradual decrease occurred until becoming similar to the adjacent bone on day 60 days after surgery. The hydroxyapatite T290800-1 accelerated the infilling of the defect provoked in the alveolar process and caused gingival inflammation and hemorrhage; however, this does not contraindicate its use.

Osteointegração da hidroxiapatita sintética no processo alveolar da mandíbula de cães: aspectos histológicos / Osteointegration of synthetic hydroxyapatite in alveolar process of mandible in dogs: histological aspects

Avaliou-se a hidroxiapatita sintética como substituto ósseo na regeneração do processo alveolar, utilizando-se 28 cães adultos hígidos, pesando entre 10 e 15kg, divididos em dois grupos. Foram criados defeitos de aproximadamente 6 x 5mm na superfície vestibular do processo alveolar até atingir a raiz do quarto pré-molar mandibular direito. Em um grupo, o defeito foi totalmente preenchido com hidroxiapatita sintética; o outro, sem tratamento, foi usado como controle. Aos 8, 15, 21, 42, 60, 90 e 120 dias, foram coletados fragmentos ósseos para a análise histológica sob microscopia óptica. Observou-se crescimento ósseo e vascular no interior dos poros de hidroxiapatita, intensa proliferação de osteoblastos e neovascularização na presença do implante. A biocompatibilidade da hidroxiapatita permitiu a sua integração com o processo alveolar por meio da formação direta de um osso lamelar. Ocorreu neoformação óssea à medida que a hidroxiapatita foi degradada.

The synthetic hydroxyapatite was evaluated as bone substitute in the regeneration of the alveolar process. Twenty-eight healthy adult dogs weighing from 10 to 15kg were divided into two groups. Defects around 6 x 5mm were provoked on the vestibular surface of the alveolar process until reaching the root of the fourth right mandibular premolar tooth. In one group the defect was totally infilled with synthetic hydroxyapatite, whereas the other untreated one was used as control. On days 8, 15, 21, 42, 60, 90 and 120, bone fragments were collected for the histological analysis under optical microscopy. The following results were observed: both bone and vascular growth inside the hydroxyapatite pores, intense osteoblast proliferations and neovascularization at the presence of the implant. The biocompatibility of the hydroxyapatite allowed its integration with the alveolar process by direct formation of a lamellar bone. The bone neoformation occurred as the hydroxyapatite was degraded.