Alterações centrais e periféricas em diferentes modelos de dor pulpar em ratos

Detalhes bibliográficos
Autor(a) principal: Filippini, Helena Fetter
Data de Publicação: 2017
Tipo de documento: Tese
Idioma: por
Título da fonte: Biblioteca Digital de Teses e Dissertações da PUC_RS
Texto Completo: http://tede2.pucrs.br/tede2/handle/tede/7184
Resumo: The central and peripheral mechanisms involved in pulpal pain are still unclear. This study compared, initially, the chronic pulpitis model evoked by the application of complete Freund's adjuvant (CFA) into the pulps of rats, with other models of pulpitis. For this purpose, the pulps of the left upper first molars of Wistar rats were accessed. There were four experimental groups according to the surgical procedure: open, closed, CFA and naïve. Behavioral changes, as well as, body weight gain and food and water consumption, were evaluated at different experimental times (1, 2, 3 and 8 days). After euthanasia, blood, brain and trigeminal ganglion (TG) samples were collected for further analysis. All experimental groups with pulp acess had a decrease in body weight gain (for up to 3 days). A reduction in food consumption was observed in all the groups with pulp exposure, on the first day. The CFA group showed a significant reduction of locomotor activity (at 1 and 3 days), which was associated with increased satellite glial cells activation in the ipsilateral TG. Activation of astrocytes in the amygdala was not affected in the studied groups. The second part of this study investigated the relevance of Toll-like receptor 4 (TLR4) activation in the acute pulpitis caused by the application of the transient receptor potential ankyrin-1 (TRPA1) mustard oil (MO), into the rat pulps. Preliminary studies were performed to compare the sensorimotor activity of the masticatory muscles with the application of lipopolysaccharide (LPS) or MO in the dental pulp. The model using MO application into the pulp of rats was selected, because of the marked increase of the electromyographic activities evoked by this agent, when compared to LPS. The animals received a intrathecal (i.t.) pre-treatment with the TLR4 LPS-RS antagonist (25 μg / 10 μl) or saline solution (10 μl) (vehicle group), 10 min before pulp application of MO. For this purpose, a stereotaxic surgery was performed to access the trigeminal subnucleus caudalis. Changes in electromyography activity (EMG) of masticatory muscles were recorded for 15 min after the MO application into the pulp. The trigeminal caudalis subnucleus was removed after euthanasia, for evaluation of TLR4 expression by western blotting analysis. Changes in EMG activity in anterior digastric muscles were significantly reduced by the intrathecal administration of LPS-RS in the trigeminal subnucleus caudalis (p ˂0.05). The data obtained suggest the participation of satellite glial cells in the chronic pulpitis model induced by CFA, as well as, TLR4 receptors in the acute pulpitis model caused by MO application into the rat pulps. Altogether, the present results shed new light on the mechanisms underlying acute and chronic tooth pulp inflammatory pain.
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spelling Campos, Maria Martha910.385.619-49http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4798056Z6916.770.090-04http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4702540A9Filippini, Helena Fetter2017-03-23T15:01:59Z2017-01-13http://tede2.pucrs.br/tede2/handle/tede/7184The central and peripheral mechanisms involved in pulpal pain are still unclear. This study compared, initially, the chronic pulpitis model evoked by the application of complete Freund's adjuvant (CFA) into the pulps of rats, with other models of pulpitis. For this purpose, the pulps of the left upper first molars of Wistar rats were accessed. There were four experimental groups according to the surgical procedure: open, closed, CFA and naïve. Behavioral changes, as well as, body weight gain and food and water consumption, were evaluated at different experimental times (1, 2, 3 and 8 days). After euthanasia, blood, brain and trigeminal ganglion (TG) samples were collected for further analysis. All experimental groups with pulp acess had a decrease in body weight gain (for up to 3 days). A reduction in food consumption was observed in all the groups with pulp exposure, on the first day. The CFA group showed a significant reduction of locomotor activity (at 1 and 3 days), which was associated with increased satellite glial cells activation in the ipsilateral TG. Activation of astrocytes in the amygdala was not affected in the studied groups. The second part of this study investigated the relevance of Toll-like receptor 4 (TLR4) activation in the acute pulpitis caused by the application of the transient receptor potential ankyrin-1 (TRPA1) mustard oil (MO), into the rat pulps. Preliminary studies were performed to compare the sensorimotor activity of the masticatory muscles with the application of lipopolysaccharide (LPS) or MO in the dental pulp. The model using MO application into the pulp of rats was selected, because of the marked increase of the electromyographic activities evoked by this agent, when compared to LPS. The animals received a intrathecal (i.t.) pre-treatment with the TLR4 LPS-RS antagonist (25 μg / 10 μl) or saline solution (10 μl) (vehicle group), 10 min before pulp application of MO. For this purpose, a stereotaxic surgery was performed to access the trigeminal subnucleus caudalis. Changes in electromyography activity (EMG) of masticatory muscles were recorded for 15 min after the MO application into the pulp. The trigeminal caudalis subnucleus was removed after euthanasia, for evaluation of TLR4 expression by western blotting analysis. Changes in EMG activity in anterior digastric muscles were significantly reduced by the intrathecal administration of LPS-RS in the trigeminal subnucleus caudalis (p ˂0.05). The data obtained suggest the participation of satellite glial cells in the chronic pulpitis model induced by CFA, as well as, TLR4 receptors in the acute pulpitis model caused by MO application into the rat pulps. Altogether, the present results shed new light on the mechanisms underlying acute and chronic tooth pulp inflammatory pain.Os mecanismos centrais e periféricos envolvidos na dor pulpar ainda não estão totalmente claros. O presente estudo comparou, inicialmente, o modelo de inflamação pulpar crônica induzido pela aplicação de adjuvante completo de Freund (CFA), em polpas de ratos, com outros modelos de pulpite. Para tanto, as polpas dos primeiros molares superiores esquerdos de ratos Wistar foram acessadas. Foram criados 4 grupos experimentais conforme a abordagem pulpar: aberto, fechado, CFA e naïve. As alterações comportamentais, bem como, o ganho de peso corporal e o consumo de água e ração, foram avaliados em diferentes tempos experimentais (1, 2, 3 e 8 dias). Após a eutanásia, amostras de sangue, cérebro e gânglio trigeminal (GT) foram coletadas para análises posteriores. Todos os grupos experimentais com exposição da polpa apresentaram diminuição do ganho de peso corporal (até o 3 º dia). A redução do consumo de ração foi observada em todos os grupos no primeiro dia. O grupo CFA mostrou redução significativa na atividade locomotora (em 1 e 3 dias), que foi associada ao aumento da ativação das células satélites da glia no GT ipsilateral. A ativação de astrócitos na amígdala não foi afetada nos grupos estudados. A segunda parte deste estudo investigou a participação do receptor Toll-like 4 (TLR4), na mediação da pulpite aguda causada pela aplicação de óleo de mostarda (OM), um agonista de receptores de potencial transitório anquirina-1 (TRPA1), em polpas de ratos. Estudos preliminares foram realizados comparando a atividade sensoriomotora dos músculos mastigatórios diante da aplicação de lipopolissacarídeo (LPS) ou OM na polpa dentária. O modelo utilizando OM na polpa de ratos foi escolhido, devido ao aumento marcante das atividades eletromiográficas (EMG), quando comparado ao LPS. Os animais foram pré-tratados por via intratecal (i.t.) com o antagonista seletivo dos receptores TLR4, o LPS-RS (25 μg/10 μl, 10 min antes da aplicação pulpar de OM. O grupo controle recebeu solução salina (10 μl) (grupo veículo), pela mesma via, no mesmo tempo de administração. Para tal, foi realizada uma cirurgia esterotáxica para permitir o acesso ao subnúcleo caudal trigeminal. As alterações das atividades de EMG dos músculos mastigatórios foram registradas durante 15 min após a aplicação de OM na polpa. O subnúcleo caudal trigeminal foi removido após a eutanásia, para avaliação da expressão de TLR4, através da análise de western-blotting. As alterações da atividade de EMG nos músculos digástricos anteriores foram significativamente reduzidas pela administração intratecal de LPS-RS no subnúcleo caudal do trigêmeo (p˂0,05), indicando um papel relevante para esses receptores nos mecanismos de sensibilização central, após a aplicação de OM. O conjunto de dados obtidos sugerem a participação das células satélites da glia no modelo de pulpite crônica induzida por CFA, bem como, dos receptores TLR4 no modelo de pulpite aguda provocada pela aplicação de OM em polpas de ratos. Essas evidências contribuem para o melhor entendimento dos mecanismos relacionados com a transmissão da dor pulpar aguda e crônica.Submitted by Caroline Xavier (caroline.xavier@pucrs.br) on 2017-03-23T15:01:59Z No. of bitstreams: 1 TES_HELENA_FETTER_FILIPPINI_PARCIAL.pdf: 1312432 bytes, checksum: dda78ca2fa347004234d72add7b38398 (MD5)Made available in DSpace on 2017-03-23T15:01:59Z (GMT). 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dc.title.por.fl_str_mv Alterações centrais e periféricas em diferentes modelos de dor pulpar em ratos
title Alterações centrais e periféricas em diferentes modelos de dor pulpar em ratos
spellingShingle Alterações centrais e periféricas em diferentes modelos de dor pulpar em ratos
Filippini, Helena Fetter
POLPA DENTÁRIA
INFLAMAÇÃO
ELETROMIOGRAFIA
ENDODONTIA
ODONTOLOGIA
CIENCIAS DA SAUDE::ODONTOLOGIA
title_short Alterações centrais e periféricas em diferentes modelos de dor pulpar em ratos
title_full Alterações centrais e periféricas em diferentes modelos de dor pulpar em ratos
title_fullStr Alterações centrais e periféricas em diferentes modelos de dor pulpar em ratos
title_full_unstemmed Alterações centrais e periféricas em diferentes modelos de dor pulpar em ratos
title_sort Alterações centrais e periféricas em diferentes modelos de dor pulpar em ratos
author Filippini, Helena Fetter
author_facet Filippini, Helena Fetter
author_role author
dc.contributor.advisor1.fl_str_mv Campos, Maria Martha
dc.contributor.advisor1ID.fl_str_mv 910.385.619-49
dc.contributor.advisor1Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4798056Z6
dc.contributor.authorID.fl_str_mv 916.770.090-04
dc.contributor.authorLattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4702540A9
dc.contributor.author.fl_str_mv Filippini, Helena Fetter
contributor_str_mv Campos, Maria Martha
dc.subject.por.fl_str_mv POLPA DENTÁRIA
INFLAMAÇÃO
ELETROMIOGRAFIA
ENDODONTIA
ODONTOLOGIA
topic POLPA DENTÁRIA
INFLAMAÇÃO
ELETROMIOGRAFIA
ENDODONTIA
ODONTOLOGIA
CIENCIAS DA SAUDE::ODONTOLOGIA
dc.subject.cnpq.fl_str_mv CIENCIAS DA SAUDE::ODONTOLOGIA
description The central and peripheral mechanisms involved in pulpal pain are still unclear. This study compared, initially, the chronic pulpitis model evoked by the application of complete Freund's adjuvant (CFA) into the pulps of rats, with other models of pulpitis. For this purpose, the pulps of the left upper first molars of Wistar rats were accessed. There were four experimental groups according to the surgical procedure: open, closed, CFA and naïve. Behavioral changes, as well as, body weight gain and food and water consumption, were evaluated at different experimental times (1, 2, 3 and 8 days). After euthanasia, blood, brain and trigeminal ganglion (TG) samples were collected for further analysis. All experimental groups with pulp acess had a decrease in body weight gain (for up to 3 days). A reduction in food consumption was observed in all the groups with pulp exposure, on the first day. The CFA group showed a significant reduction of locomotor activity (at 1 and 3 days), which was associated with increased satellite glial cells activation in the ipsilateral TG. Activation of astrocytes in the amygdala was not affected in the studied groups. The second part of this study investigated the relevance of Toll-like receptor 4 (TLR4) activation in the acute pulpitis caused by the application of the transient receptor potential ankyrin-1 (TRPA1) mustard oil (MO), into the rat pulps. Preliminary studies were performed to compare the sensorimotor activity of the masticatory muscles with the application of lipopolysaccharide (LPS) or MO in the dental pulp. The model using MO application into the pulp of rats was selected, because of the marked increase of the electromyographic activities evoked by this agent, when compared to LPS. The animals received a intrathecal (i.t.) pre-treatment with the TLR4 LPS-RS antagonist (25 μg / 10 μl) or saline solution (10 μl) (vehicle group), 10 min before pulp application of MO. For this purpose, a stereotaxic surgery was performed to access the trigeminal subnucleus caudalis. Changes in electromyography activity (EMG) of masticatory muscles were recorded for 15 min after the MO application into the pulp. The trigeminal caudalis subnucleus was removed after euthanasia, for evaluation of TLR4 expression by western blotting analysis. Changes in EMG activity in anterior digastric muscles were significantly reduced by the intrathecal administration of LPS-RS in the trigeminal subnucleus caudalis (p ˂0.05). The data obtained suggest the participation of satellite glial cells in the chronic pulpitis model induced by CFA, as well as, TLR4 receptors in the acute pulpitis model caused by MO application into the rat pulps. Altogether, the present results shed new light on the mechanisms underlying acute and chronic tooth pulp inflammatory pain.
publishDate 2017
dc.date.accessioned.fl_str_mv 2017-03-23T15:01:59Z
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dc.publisher.department.fl_str_mv Faculdade de Odontologia
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