Avaliação do efeito do transplante de células-tronco mesenquimais derivadas de medula óssea em modelo murino de neuropatia periférica diabética

Submitted by Ana Maria Fiscina Sampaio ([email protected]) on 2015-03-06T16:58:42Z No. of bitstreams: 1 Afrânio Ferreira Evangelista Avaliação...2014.pdf: 2792913 bytes, checksum: 154973247ed482dbfeac342e4b641d7c (MD5)Approved for entry into archive by Ana Maria Fiscina Sampaio ([email protected]) on 2015-03-06T17:00:06Z (GMT) No. of bitstreams: 1 Afrânio Ferreira Evangelista Avaliação...2014.pdf: 2792913 bytes, checksum: 154973247ed482dbfeac342e4b641d7c (MD5)Approved for entry into archive by Ana Maria Fiscina Sampaio ([email protected]) on 2015-03-06T17:01:18Z (GMT) No. of bitstreams: 1 Afrânio Ferreira Evangelista Avaliação...2014.pdf: 2792913 bytes, checksum: 154973247ed482dbfeac342e4b641d7c (MD5)Made available in DSpace on 2015-03-06T17:01:18Z (GMT). No. of bitstreams: 1 Afrânio Ferreira Evangelista Avaliação...2014.pdf: 2792913 bytes, checksum: 154973247ed482dbfeac342e4b641d7c (MD5) Previous issue date: 2014Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, BrasilO diabetes é uma doença de alta prevalência que, frequentemente, induz o comprometimento do sistema nervoso periférico. Na neuropatia diabética periférica, os sintomas mais encontrados são os sensitivos, no qual a dor neuropática, condição crônica caracterizada por alodinia e hiperalgesia, é a mais debilitante. Esta, prejudica a qualidade de vida do paciente, sendo muitas vezes não responsiva aos métodos farmacológicos convencionais de tratamento. Diante desse panorama, o desenvolvimento de novas abordagens terapêuticas que possuam ação efetiva neste tipo de dor é de grande relevância. O uso da terapia celular no tratamento de lesões do sistema nervoso tem demonstrado resultados promissores e o potencial terapêutico de células-tronco na neuropatia experimental tem sido proposto. Neste estudo, avaliou-se o efeito de células-tronco mesenquimais derivadas da medula óssea (CMsMO) na neuropatia diabética periférica estabelecida em modelo experimental de diabetes induzido por estreptozotocina (ETZ). Quatro semanas após a indução do modelo por ETZ (80 mg/kg; ip; 3 dias consecutivos), os animais receberam uma administração endovenosa de CMsMO (1 x 106) ou veículo. O tratamento com gabapentina (30 mg/kg; v.o. a cada 12 horas durante seis dias consecutivos) foi usado como padrão ouro. Os limiares nociceptivos térmico e mecânico foram avaliados durante todo o período experimental (90 dias), pelos métodos de hargreaves e von Frey. A avaliação da função motora foi realizada pelo teste de rota-rod. Em diferentes tempos e para todos os grupos experimentais, foram realizadas coletas de segmentos da medula espinal (L4-L5) para dosagem de citocinas por ELISA e segmentos do nervo isquiático foram também coletados para avaliação de alterações morfológicas por microscopia óptica e eletrônica de transmissão. Os dados comportamentais demonstraram que o tratamento com CMsMO reduziu a mecanoalodinia e a hipoalgesia térmica, levando os limiares nociceptivos de animais neuropáticos a níveis similares aos de animais não neuropáticos. Do mesmo modo, a administração de CMsMO normalizou a função motora dos animais neuropáticos. Dados de microscopia mostraram que animais neuropáticos apresentaram atrofia axonal, redução do número de fibras mielínicas e aparente redução do numero de fibras amielínicas no nervo isquiático. Animais neuropáticos tratados com CMsMO tiveram menor ocorrência de atrofia axonal e não apresentaram redução do numero de fibras mielínicas ou amielínicas, em relação aos neuropáticos tratados com salina. Além disso, animais neuropáticos tratados com CMsMO apresentaram menores níveis espinais de IL-1β e TNF-α, e maiores de IL-10 e TGF-β, em relação aos animais neuropáticos não tratados. Esse conjunto de resultados indica que CMsMO produzem efeito antinociceptivo duradouro na neuropatia diabética, seguido de modificações no padrão fisiopatológico da doença, o que aponta a terapia celular como uma interessante alternativa para o controle da neuropatia diabética periférica dolorosa.Diabetes is a highly prevalent disease which frequently compromises the peripheral nervous system. In peripheral diabetic neuropathy, the most frequent symptoms are sensitive, in which the neuropathic pain, chronic condition characterized by allodynia and hyperalgesia, is the most debilitating. Neuropathic pain affects the quality of patients’ lives, and is often not responsive to pharmacological conventional treatment methods. Against this background, the development of new therapeutic approaches that have an effective action in this type of pain is of great importance. The use of cell therapy in the treatment of lesions in the nervous system has shown promising results and the therapeutic potential of stem cells in experimental neuropathy has been proposed. In this study, we evaluated the effect of mesenchymal stem cells derived from bone marrow (CMsMO) in peripheral diabetic neuropathy established in experimental model of streptozotocin (STZ) induced diabetes in mice. Four weeks after the induction of the model by administration of STZ (80 mg/kg, ip; 3 days) the animals received an CMsMO by intravenous administration (1x106) or vehicle. The treatment with gabapentin (30 mg/kg, orally every 12 hours for six days) was used as the gold standard. The thermal and mechanical nociceptive thresholds were assessed throughout the entire experimental period (90 days), using Hargreaves and von Frey methods, respectively. Motor function evaluation of was conducted using the rotarod test. At different times, were analyzes conducted in spinal cord segments (L4-L5) to determine cytokines profile by ELISA. Sciatic nerve segments were also collected for evaluation of morphological changes by optical and electron transmission microscopy. According to the behavioral data, the CMsMO treatment reduced the mecanoalodinia and the thermal hypoalgesia, leading nociceptive thresholds of neuropathic animals to levels similar to those of non-neuropathic animals. Similarly, CMsMO administration normalized motor function of neuropathic animals. Microscopy data demonstrated that neuropathic animals had axonal atrophy and an apparent decrease of the number of myelinated fibers as well a reduction in the number of unmyelinated fibers in the sciatic nerve, but neuropathic animals treated with CMsMO had a lower incidence of axonal atrophy, showed no decrease in the number of myelinated fibers and no apparent decrease in the amount of unmyelinated fibers in relation to neuropathics treated with saline. Furthermore, neuropathic animals treated with CMsMO presented lower levels of spinal IL-1β and higher levels of TNF-α, and IL-10 and TGF-β compared to neuropathic animals that received saline. These data indicate that CMsMO produces a lasting analgesic effect in diabetic neuropathy, followed by changes in the pathophysiological disease pattern, which indicates cell therapy as an interesting alternative for the control of painful peripheral diabetic neuropathy

Mecanismos envolvidos no efeito terapêutico de células mesenquimais de medula óssea em modelo experimental de neuropatia diabética sensorial

Submitted by Ana Maria Fiscina Sampaio ([email protected]) on 2020-03-03T11:37:53Z No. of bitstreams: 1 Afranio Ferreira Evangelista. Mecanismos envolvidos..2019.pdf: 15708019 bytes, checksum: 22fc1807a4ae834d7e4415855c2ee624 (MD5)Approved for entry into archive by Ana Maria Fiscina Sampaio ([email protected]) on 2020-03-03T11:38:08Z (GMT) No. of bitstreams: 1 Afranio Ferreira Evangelista. Mecanismos envolvidos..2019.pdf: 15708019 bytes, checksum: 22fc1807a4ae834d7e4415855c2ee624 (MD5)Made available in DSpace on 2020-03-03T11:38:08Z (GMT). No. of bitstreams: 1 Afranio Ferreira Evangelista. Mecanismos envolvidos..2019.pdf: 15708019 bytes, checksum: 22fc1807a4ae834d7e4415855c2ee624 (MD5) Previous issue date: 2019O presente trabalho foi realizado com apoio da Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - Brasil (CAPES) – Código de Financiamento 001. FIOCRUZ. CNPq. FAPESB.Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil.INTRODUÇÃO: A neuropatia diabética sensorial (NDS) é uma das mais comuns complicações do diabetes que se manifesta por distúrbios sensitivos crônicos como dor neuropática e perda de sensibilidade, os quais contribuem para o desenvolvimento do pé diabético, podendo resultar em ulceração e amputação do membro afetado. Diante deste cenário, em função de suas propriedades regenerativas e analgésicas, células mesenquimais de medula óssea (CMsMO) têm sido consideradas promissoras no tratamento das neuropatias em relação a terapêutica atual disponível. OBJETIVO: Investigar os possíveis mecanismos envolvidos no efeito terapêutico das CMsMO em modelo murinho de NDS. MÉTODOS: Quatro semanas após a indução do modelo por estreptozotocina (STZ), camundongos C57Bl/6 (CEUA L-IGM-025/11) receberam uma administração endovenosa de CMsMO (1x106), salina ou gabapentina (70 mg/kg, v.o., 12h/12h por 6 dias) e os limiares nociceptivos térmico (hargreaves) e mecânico (filamentos de von Frey) foram avaliados por 12 semanas. Na 12ª semana, segmentos do nervo isquiático foram coletados para avaliações histológicas por microscopia óptica e eletrônica de transmissão. Amostras de nervo isquiático, medula espinal, gânglio da raiz dorsal (GRD), baço e fígado foram recolhidas para pesquisa das CMsMO transplantadas por PCR quantitativo em Tempo Real (RT-qPCR), assim como a porção L4-L5 da medula espinal foi coletada para análise da expressão de fatores antioxidantes e quantificação de produtos do estresse oxidativo. O perfil de ativação de micróglia, astrócitos e níveis de expressão de galectina-3 na medula espinal foram avaliados por imunofluorescência. Para investigar a hipótese da ação parácrina, o efeito do meio condicionado obtido do cultivo de CMsMO (MC-CMsMO) ou vesículas extracelulares derivadas de CMsMO (VEs-CMsMO) sobre o limiar nociceptivo foi avaliado e VEs-CMsMO foram caracterizadas quanto a sua morfologia, tamanho, quantidade e conteúdo proteico. RESULTADOS: O tratamento com CMsMO reduziu a alodinia mecâ nica e a hipoalgesia térmica, igualando os limiares nociceptivos de animais neuropáticos àqueles do grupo naive. Por outro lado, o tratamento com gabapentina (70mg/kg) reduziu a alodinia e a hipoalgesia somente durante o protocolo de administraçã o do fármaco. Análises por microscopia mostraram que animais com neuropatia apresentaram atrofia axonal, reduçã o do número de fibras mielínicas, atipia mitocondrial e redução da área e densidade das fibras amielínicas no nervo isquiático e que o transplante com CMsMO preveniu o desenvolvimento de tais alterações histológicas no nervo, bem como o aumento dos níveis de expressã o de fatores antioxidantes, nitrito e MDA na medula de animais com NDS. Animais neuropáticos apresentaram aumento no número de astrócitos e micróglia ativados, assim como expressã o de galectina-3 na medula espinal. Este efeito foi inibido pelo transplante de CMsMO, evidenciando açã o moduladora das CMsMO na neuroinflamaçã o espinal. O rastreio das células transplantadas demonstrou que estas possuem tendência em migrar para órgãos filtrantes e ainda assim produzir efeito, evidenciando uma açã o parácrina. Em linha com esta hipótese, a infusã o do MC-CMsMO em animais diabéticos conseguiu reverter os parâmetros comportamentais da neuropatia sensorial de forma semelhante ao tratamento com CMsMO. Resultados da caracterizaçã o das VEs-CMsMO mostraram vesículas com tamanho entre 100-500nm, porém a análise do seu proteoma revelou-se inconclusiva. Contudo, a administraçã o de VEs-CMsMO foi capaz de produzir efeito antinociceptivo, levando os limiares nociceptivos de animais com neuropatia a níveis similares ao de animais saudáveis. CONCLUSÃO: Este conjunto de resultados mostram que o transplante de CMsMO ou infusão do MC-CMsMO ou VEs-CMsMO foram capazes de reverter os parâmetros comportamentais indicativos de dor neuropática e que, além dos efeitos neuroprotetores de CMsMO no nervo periférico, sua ação parácrina pode ser responsável pela modulação da neuroinflamação espinal, podendo esta ser considerada como um importante mecanismo pelo qual as células mesenquimais induzem efeitos terapêuticos durante a neuropatia diabética sensorial.INTRODUCTION: Sensory diabetic neuropathy (SDN) is one of the most common complications of diabetes manifested by chronic sensory disorders such as neuropathic pain and loss of sensation, which contribute to the development of the diabetic foot, which may result in ulceration and amputation of the affected limb. Given this scenario, due to their regenerative and analgesic properties, bone marrow mesenchymal cells (BM-MSC) have been considered promising in the treatment of neuropathies in relation to current available therapy. AIM: To investigate the possible mechanisms involved in the therapeutic effect of BM-MSC in a murine SDN model. METHODS: Four weeks after streptozotocin (STZ) model induction, C57Bl/6 mice (CEUA L-IGM-025/11) received intravenous administration of BM-MSC (1x106), saline or gabapentin (70 mg / kg, vo, 12h / 12h for 6 days) and the thermal (hargreaves) and mechanical (von Frey filaments) nociceptive thresholds were evaluated for 12 weeks. At week 12, sciatic nerve segments were collected for histological evaluations by light and transmission electron microscopy. Samples of sciatic nerve, spinal cord, dorsal root ganglion (DRG), spleen, and liver were collected for quantitative Real-Time PCR (RT-qPCR) transplanted BM-MSC, as well as the L4-L5 portion of the spinal cord was collected. for analysis of the expression of antioxidant factors and quantification of oxidative stress products. The activation profile of microglia, astrocytes and levels of galectin-3 expression in the spinal cord were evaluated by immunofluorescence. To investigate the hypothesis of paracrine action, the effect of conditioned media obtained from culturing BM-MSC (CM-BM-MSC) or extracellular vesicles derived from BM-MSC (EVs-BM-MSC) on the nociceptive threshold was evaluated and VEs- BM-MSC were characterized for morphology. , size, quantity and protein content.RESULTS: BM-MSC treatment reduced mechanical allodynia and thermal hypoalgesia by equalizing the nociceptive thresholds of neuropathic animals to those in the naive group. Conversely, gabapentin treatment (70mg/kg) reduced allodynia and hypoalgesia only during the drug administration protocol. Microscopic analysis showed that animals with neuropathy had axonal atrophy, reduced number of myelin fibers, mitochondrial atypia, and reduced area and density of myelinic fibers in the sciatic nerve and that transplantation with BM-MSC prevented the development of such histological changes in the nerve, as well as as increased levels of expression of antioxidant factors, nitrite and MDA in the marrow of animals with SDN. Neuropathic animals showed increased number of activated astrocytes and microglia, as well as expression of galectin-3 in the spinal cord. This effect was inhibited by BM-MSC transplantation, showing modulating action of BM-MSC on spinal neuroinflammation. Screening of transplanted cells showed that they have a tendency to migrate to filtering organs and still produce effect, showing a paracrine action. In line with this hypothesis, CM-BM-MSC infusion in diabetic animals was able to reverse the behavioral parameters of sensory neuropathy similarly to BM-MSC treatment. Results of the characterization of the EVs-BMMSC showed vesicles with size between 100-500nm, but the analysis of their proteome proved inconclusive. However, the administration of EVs-BM-MSC was able to produce antinociceptive effect, bringing nociceptive thresholds of animals with neuropathy to similar levels to healthy animals. CONCLUSION: This set of results show that BM-MSC transplantation, CM-BM-MSC or EVs-BM-MSC infusion were able to reverse behavioral parameters indicative of neuropathic pain and that, in addition to the neuroprotective effects of BM-MSC on the peripheral nerve, its paracrine action may be responsible. by modulation of spinal neuroinflammation, which can be considered as an important mechanism by which mesenchymal cells induce therapeutic effects during sensory diabetic neuropathy

Cell-free therapy: a neuroregenerative approach to sensory neuropathy?

Submitted by Ana Maria Fiscina Sampaio ([email protected]) on 2019-08-07T13:42:17Z No. of bitstreams: 1 Villareal, C.F. Cell-free therapy... 2019.pdf: 202951 bytes, checksum: 68bde7428021452f98a33aac1002aa7c (MD5)Approved for entry into archive by Ana Maria Fiscina Sampaio ([email protected]) on 2019-08-07T14:06:40Z (GMT) No. of bitstreams: 1 Villareal, C.F. Cell-free therapy... 2019.pdf: 202951 bytes, checksum: 68bde7428021452f98a33aac1002aa7c (MD5)Made available in DSpace on 2019-08-07T14:06:40Z (GMT). No. of bitstreams: 1 Villareal, C.F. Cell-free therapy... 2019.pdf: 202951 bytes, checksum: 68bde7428021452f98a33aac1002aa7c (MD5) Previous issue date: 2019Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil.Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil.Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil / Universidade Federal da Bahia. Faculdade de Farmácia. Salvador, BA, Brasil / Instituto Nacional de Ciência e Tecnologia. Instituto de Investigação em Imunologia. São Paulo, SP, Brasil

Antinociceptive properties of physalins from Physalis angulata.

Submitted by Ana Maria Fiscina Sampaio ([email protected]) on 2015-03-11T19:17:13Z No. of bitstreams: 1 Lima MS Antinocicptive....pdf: 992187 bytes, checksum: 7a499c23ff111fc40f8a4d978d9113e0 (MD5)Approved for entry into archive by Ana Maria Fiscina Sampaio ([email protected]) on 2015-03-11T19:17:22Z (GMT) No. of bitstreams: 1 Lima MS Antinocicptive....pdf: 992187 bytes, checksum: 7a499c23ff111fc40f8a4d978d9113e0 (MD5)Approved for entry into archive by Ana Maria Fiscina Sampaio ([email protected]) on 2015-03-16T17:10:13Z (GMT) No. of bitstreams: 1 Lima MS Antinocicptive....pdf: 992187 bytes, checksum: 7a499c23ff111fc40f8a4d978d9113e0 (MD5)Made available in DSpace on 2015-03-16T17:10:14Z (GMT). No. of bitstreams: 1 Lima MS Antinocicptive....pdf: 992187 bytes, checksum: 7a499c23ff111fc40f8a4d978d9113e0 (MD5) Previous issue date: 2014Universidade Federal da Bahia. Faculdade de Farmácia. Salvador, BA, BrasilFundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, BrasilFundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, BrasilFundação Oswaldo Cruz. FarManguinhos. Rio de Janeiro, RJ, BrasilFundação Oswaldo Cruz. FarManguinhos. Rio de Janeiro, RJ, BrasilFundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, Brasil / Hospital São Rafael. Centro de Biotecnologia e Terapia Celular. Salvador, BA, BrasilUniversidade Federal da Bahia. Faculdade de Farmácia. Salvador, BA, Brasil /Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, BrasilPain is the most common reason a patient sees a physician. Nevertheless, the use of typical painkillers is not completely effective in controlling all pain syndromes; therefore further attempts have been made to develop improved analgesic drugs. The present study was undertaken to evaluate the antinociceptive properties of physalins B (1), D (2), F (3), and G (4) isolated from Physalis angulata in inflammatory and centrally mediated pain tests in mice. Systemic pretreatment with 1-4 produced dose-related antinociceptive effects on the writhing and formalin tests, traditional screening tools for the assessment of analgesic drugs. On the other hand, only 3 inhibited inflammatory parameters such as hyperalgesia, edema, and local production of TNF-α following induction with complete Freund's adjuvant. Treatment with 1, 3, and 4 produced an antinociceptive effect on the tail flick test, suggesting a centrally mediated antinociception. Reinforcing this idea, 2-4 enhanced the mice latency reaction time during the hot plate test. Mice treated with physalins did not demonstrate motor performance alterations. These results suggest that 1-4 present antinociceptive properties associated with central, but not anti-inflammatory, events and indicate a new pharmacological property of physalins

Conditioned Medium of Bone Marrow-Derived Mesenchymal Stromal Cells as a Therapeutic Approach to Neuropathic Pain: A Preclinical Evaluation

Submitted by Ana Maria Fiscina Sampaio ([email protected]) on 2018-08-14T12:53:03Z No. of bitstreams: 1 Gama KB Conditioned Medium of Bone Marrow....pdf: 1608380 bytes, checksum: 38bd383f4cde0cfd9b310fcd2fbda626 (MD5)Approved for entry into archive by Ana Maria Fiscina Sampaio ([email protected]) on 2018-08-14T13:28:57Z (GMT) No. of bitstreams: 1 Gama KB Conditioned Medium of Bone Marrow....pdf: 1608380 bytes, checksum: 38bd383f4cde0cfd9b310fcd2fbda626 (MD5)Made available in DSpace on 2018-08-14T13:28:57Z (GMT). No. of bitstreams: 1 Gama KB Conditioned Medium of Bone Marrow....pdf: 1608380 bytes, checksum: 38bd383f4cde0cfd9b310fcd2fbda626 (MD5) Previous issue date: 2018FAPESB (Grant no. DTE 0046/2011) and CNPq (Grant no. 445547/2014-6).Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil / Universidade Federal da Bahia. Faculdade de Farmácia. Salvador, BA, BrasilUniversidade Federal da Bahia. Faculdade de Farmácia. Salvador, BA, BrasilFundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, BrasilFundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil / Hospital São Rafael. Centro de Biotecnologia e Terapia Celular. Salvador, BA, BrasilHospital São Rafael. Centro de Biotecnologia e Terapia Celular. Salvador, BA, BrasilHospital São Rafael. Centro de Biotecnologia e Terapia Celular. Salvador, BA, Brasil / National Institute of Science and Technology for Regenerative Medicine. Rio de Janeiro, RJ, BrazilFundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil / Hospital São Rafael. Centro de Biotecnologia e Terapia Celular. Salvador, BA, Brasil / National Institute of Science and Technology for Regenerative Medicine. Rio de Janeiro, RJ, BrazilFundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil / Universidade Federal da Bahia. Faculdade de Farmácia. Salvador, BA, BrasilNeuropathic pain is a type of chronic pain caused by injury or dysfunction of the nervous system, without effective therapeutic approaches. Mesenchymal stromal cells (MSCs), through their paracrine action, have great potential in the treatment of this syndrome. In the present study, the therapeutic potential of MSC-derived conditioned medium (CM) was investigated in a mouse model of neuropathic pain induced by partial sciatic nerve ligation (PSL). PSL mice were treated by endovenous route with bone marrow-derived MSCs (1 × 106), CM, or vehicle. Gabapentin was the reference drug. Twelve hours after administration, neuropathic mice treated with CM exhibited an antinociceptive effect that was maintained throughout the evaluation period. MSCs also induced nonreversed antinociception, while gabapentin induced short-lasting antinociception. The levels of IL-1β, TNF-α, and IL-6 were reduced, while IL-10 was enhanced on sciatic nerve and spinal cord by treatment with CM and MSCs. Preliminary analysis of the CM secretome revealed the presence of growth factors and cytokines likely involved in the antinociception. In conclusion, the CM, similar to injection of live cells, produces a powerful and long-lasting antinociceptive effect on neuropathic pain, which is related with modulatory properties on peripheral and central levels of cytokines involved with the maintenance of this syndrome

Unveiling Targets for Treating Postoperative Pain: The Role of the TNF-α/p38 MAPK/NF-κB/Nav1.8 and Nav1.9 Pathways in the Mouse Model of Incisional Pain

Although the mouse model of incisional pain is broadly used, the mechanisms underlying plantar incision-induced nociception are not fully understood. This work investigates the role of Nav1.8 and Nav1.9 sodium channels in nociceptive sensitization following plantar incision in mice and the signaling pathway modulating these channels. A surgical incision was made in the plantar hind paw of male Swiss mice. Nociceptive thresholds were assessed by von Frey filaments. Gene expression of Nav1.8, Nav1.9, TNF-α, and COX-2 was evaluated by Real-Time PCR in dorsal root ganglia (DRG). Knockdown mice for Nav1.8 and Nav1.9 were produced by antisense oligodeoxynucleotides intrathecal treatments. Local levels of TNF-α and PGE2 were immunoenzymatically determined. Incised mice exhibited hypernociception and upregulated expression of Nav1.8 and Nav1.9 in DRG. Antisense oligodeoxynucleotides reduced hypernociception and downregulated Nav1.8 and Nav1.9. TNF-α and COX-2/PGE2 were upregulated in DRG and plantar skin. Inhibition of TNF-α and COX-2 reduced hypernociception, but only TNF-α inhibition downregulated Nav1.8 and Nav1.9. Antagonizing NF-κB and p38 mitogen-activated protein kinase (MAPK), but not ERK or JNK, reduced both hypernociception and hyperexpression of Nav1.8 and Nav1.9. This study proposes the contribution of the TNF-α/p38/NF-κB/Nav1.8 and Nav1.9 pathways to the pathophysiology of the mouse model of incisional pain

Bone marrow-derived mesenchymal stem/stromal cells reverse the sensorial diabetic neuropathy via modulation of spinal neuroinflammatory cascades

Submitted by Ana Maria Fiscina Sampaio ([email protected]) on 2018-08-14T12:25:36Z No. of bitstreams: 1 Evangelista AF Bone marrow-derived mesenchymal ....pdf: 3920676 bytes, checksum: c9784ec89d0678d1be3c53cf120dd471 (MD5)Approved for entry into archive by Ana Maria Fiscina Sampaio ([email protected]) on 2018-08-14T12:46:12Z (GMT) No. of bitstreams: 1 Evangelista AF Bone marrow-derived mesenchymal ....pdf: 3920676 bytes, checksum: c9784ec89d0678d1be3c53cf120dd471 (MD5)Made available in DSpace on 2018-08-14T12:46:12Z (GMT). No. of bitstreams: 1 Evangelista AF Bone marrow-derived mesenchymal ....pdf: 3920676 bytes, checksum: c9784ec89d0678d1be3c53cf120dd471 (MD5) Previous issue date: 2018National Institutes of Health guide for the care and use of Laboratory animals (NIH, 8023) and the Institutional Animal Care and Use Committee FIOCRUZ (CPqGM 025/2011)Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, BrasilFundação Oswaldo Cruz. Instituto Oswaldo Cruz. Rio de Janeiro, RJ, BrasilFederal University of Bahia. Pharmacy College. Salvador, BA, BrasilSão Rafael Hospital. Center of Biotechnology and Cell Therapy. Salvador, BA, BrazilFederal University of Recôncavo of Bahia. Feira de Santana, BA, BrazilSão Rafael Hospital. Center of Biotechnology and Cell Therapy. Salvador, BA, BrazilSão Rafael Hospital. Center of Biotechnology and Cell Therapy. Salvador, BA, BrazilFundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil / São Rafael Hospital. Center of Biotechnology and Cell Therapy. Salvador, BA, BrazilFundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil / Federal University of Bahia. Pharmacy College. Salvador, BA, BrasilDiabetic neuropathy (DN) is a frequent and debilitating manifestation of diabetes mellitus, to which there are no effective therapeutic approaches. Mesenchymal stem/stromal cells (MSC) have a great potential for the treatment of this syndrome, possibly through regenerative actions on peripheral nerves. Here, we evaluated the therapeutic effects of MSC on spinal neuroinflammation, as well as on ultrastructural aspects of the peripheral nerve in DN-associated sensorial dysfunction. Methods: C57Bl/6 mice were treated with bone marrow-derived MSC (1 × 106), conditioned medium from MSC cultures (CM-MSC) or vehicle by endovenous route following the onset of streptozotocin (STZ)-induced diabetes. Paw mechanical and thermal nociceptive thresholds were evaluated by using von Frey filaments and Hargreaves test, respectively. Morphological and morphometric analysis of the sciatic nerve was performed by light microscopy and transmission electron microscopy. Mediators and markers of neuroinflammation in the spinal cord were measured by radioimmunoassay, real-time PCR, and immunofluorescence analyses. Results: Diabetic mice presented behavioral signs of sensory neuropathy, mechanical allodynia, and heat hypoalgesia, which were completely reversed by a single administration of MSC or CM-MSC. The ultrastructural analysis of the sciatic nerve showed that diabetic mice exhibited morphological and morphometric alterations, considered hallmarks of DN, such as degenerative changes in axons and myelin sheath, and reduced area and density of unmyelinated fibers. In MSC-treated mice, these structural alterations were markedly less commonly observed and/or less pronounced. Moreover, MSC transplantation inhibited multiple parameters of spinal neuroinflammation found in diabetic mice, causing the reduction of activated astrocytes and microglia, oxidative stress signals, galectin-3, IL-1β, and TNF-α production. Conversely, MSC increased the levels of anti-inflammatory cytokines, IL-10, and TGF-β. Conclusions: The present study described the modulatory effects of MSC on spinal cord neuroinflammation in diabetic mice, suggesting new mechanisms by which MSC can improve DN

Arquivos Brasileiros de Psiquiatria, Neurologia e Ciências Afins: uma fonte com muita história Arquivos Brasileiros de Psiquiatria, Neurologia e Ciências Afins: a source rich in history

Surgido em 1905, o Arquivos Brasileiros de Psiquiatria, Neurologia e Ciências Afins foi o primeiro periódico brasileiro especializado na área. Com o nome de Arquivos Brasileiros de Psiquiatria, Neurologia e Medicina Legal, tornou-se, em 1908, veículo de divulgação da Sociedade com o mesmo nome. A partir de 1919 foi publicado como Arquivos Brasileiros de Neuriatria e Psiquiatria. O periódico traz as propostas para a psiquiatria brasileira lideradas por Juliano Moreira, diretor do Hospício Nacional e da Assistência a Alienados do Distrito Federal. Por veicular trabalhos de profissionais do Hospício Nacional, é fonte fundamental para a investigação dos processos diagnósticos e das práticas clínicas e terapêuticas do período. Apresenta-se aqui uma seleção de artigos publicados na revista entre 1905 e 1930, ano da aposentadoria de Moreira.<br>Launched in 1905, the Arquivos Brasileiros de Psiquiatria, Neurologia e Ciências Afins was the first Brazilian periodical specialized in psychiatry, neurology, and related sciences. In 1908, under the new name of Arquivos Brasileiros de Psiquiatria, Neurologia e Medicina Legal, it became the journal of the Society of Psychiatry, Neurology, and Forensic Medicine. Starting in 1919, it was published under the title Arquivos Brasileiros de Neuriatria e Psiquiatria. The periodical presents proposals put forward for this field in Brazil, under the leadership of Juliano Moreira, director of both the National Asylum and of Federal District Assistance for the Insane (Assistência a Alienados do Distrito Federal). Because the journal published articles by healthcare providers from the National Asylum, it is a vital source for researchers of the era's diagnostic processes and clinical and therapeutic practices. This selection of texts was published in its pages between 1905 and 1930, the year Moreira retired