Characterization of the effects of antiepileptic drugs on bone metabolism: in vitro studies with human osteoblastic and osteoclastic cells

Detalhes bibliográficos
Autor(a) principal: Rocha, Sara
Data de Publicação: 2013
Tipo de documento: Dissertação
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/10400.22/3470
Resumo: Bone is constantly being molded and shaped by the action of osteoclasts and osteoblasts. A proper equilibrium between both cell types metabolic activities is required to ensure an adequate skeletal tissue structure, and it involves resorption of old bone and formation of new bone tissue. It is reported that treatment with antiepileptic drugs (AEDs) can elicit alterations in skeletal structure, in particular in bone mineral density. Nevertheless, the knowledge regarding the effects of AEDs on bone cells are still scarce. In this context, the aim of this study was to investigate the effects of five different AEDs on human osteoclastic, osteoblastic and co-cultured cells. Osteoclastic cell cultures were established from precursor cells isolated from human peripheral blood and were characterized for tartrate-resistant acid phosphatase (TRAP) activity, number of TRAP+ multinucleated cells, presence of cells with actin rings and expressing vitronectin and calcitonin receptors and apoptosis rate. Also, the involvement of several signaling pathways on the cellular response was addressed. Osteoblastic cell cultures were obtained from femur heads of patients (25-45 years old) undergoing orthopaedic surgery procedures and were then studied for cellular proliferation/viability, ALP activity, histochemical staining of ALP and apoptosis rate. Also the expression of osteoblast-related genes and the involvement of some osteoblastogenesis-related signalling pathways on cellular response were addressed. For co-cultured cells, osteoblastic cells were firstly seeded and cultured. After that, PBMC were added to the osteoblastic cells and co-cultures were evaluated using the same osteoclast and osteoblast parameters mentioned above for the corresponding isolated cell. Cell-cultures were maintained in the absence (control) or in the presence of different AEDs (carbamazepine, gabapentin, lamotrigine, topiramate and valproic acid). All the tested drugs were able to affect osteoclastic and osteoblastic cells development, although with different profiles on their osteoclastogenic and osteoblastogenic modulation properties. Globally, the tendency was to inhibit the process. Furthermore, the signaling pathways involved in the process also seemed to be differently affected by the AEDs, suggesting that the different drugs may affect osteoclastogenesis and/or osteoblastogenesis through different mechanisms. In conclusion, the present study showed that the different AEDs had the ability to directly and indirectly modulate bone cells differentiation, shedding new light towards a better understanding of how these drugs can affect bone tissue.
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spelling Characterization of the effects of antiepileptic drugs on bone metabolism: in vitro studies with human osteoblastic and osteoclastic cellsBone remodelingOsteoclastic cellsOsteoblastic cellsOsteoclastogenesisOsteoblastogenesisAntiepileptic drugsEpilepsyRemodelação ósseaOsteoclastosOsteoblastosOsteoclastogéneseOsteoblastogéneseAntiepilépticosEpilepsiaBone is constantly being molded and shaped by the action of osteoclasts and osteoblasts. A proper equilibrium between both cell types metabolic activities is required to ensure an adequate skeletal tissue structure, and it involves resorption of old bone and formation of new bone tissue. It is reported that treatment with antiepileptic drugs (AEDs) can elicit alterations in skeletal structure, in particular in bone mineral density. Nevertheless, the knowledge regarding the effects of AEDs on bone cells are still scarce. In this context, the aim of this study was to investigate the effects of five different AEDs on human osteoclastic, osteoblastic and co-cultured cells. Osteoclastic cell cultures were established from precursor cells isolated from human peripheral blood and were characterized for tartrate-resistant acid phosphatase (TRAP) activity, number of TRAP+ multinucleated cells, presence of cells with actin rings and expressing vitronectin and calcitonin receptors and apoptosis rate. Also, the involvement of several signaling pathways on the cellular response was addressed. Osteoblastic cell cultures were obtained from femur heads of patients (25-45 years old) undergoing orthopaedic surgery procedures and were then studied for cellular proliferation/viability, ALP activity, histochemical staining of ALP and apoptosis rate. Also the expression of osteoblast-related genes and the involvement of some osteoblastogenesis-related signalling pathways on cellular response were addressed. For co-cultured cells, osteoblastic cells were firstly seeded and cultured. After that, PBMC were added to the osteoblastic cells and co-cultures were evaluated using the same osteoclast and osteoblast parameters mentioned above for the corresponding isolated cell. Cell-cultures were maintained in the absence (control) or in the presence of different AEDs (carbamazepine, gabapentin, lamotrigine, topiramate and valproic acid). All the tested drugs were able to affect osteoclastic and osteoblastic cells development, although with different profiles on their osteoclastogenic and osteoblastogenic modulation properties. Globally, the tendency was to inhibit the process. Furthermore, the signaling pathways involved in the process also seemed to be differently affected by the AEDs, suggesting that the different drugs may affect osteoclastogenesis and/or osteoblastogenesis through different mechanisms. In conclusion, the present study showed that the different AEDs had the ability to directly and indirectly modulate bone cells differentiation, shedding new light towards a better understanding of how these drugs can affect bone tissue.O tecido ósseo sofre remodelação constante por ação dos osteoclastos e osteoblastos. Um equilíbrio adequado entre as atividades metabólicas de ambas as células torna-se essencial para garantir uma estrutura apropriada do tecido esquelético, e envolve a reabsorção de osso velho e consequente formação de novo tecido ósseo. Alterações na estrutura esquelética, em particular na densidade mineral óssea, por parte de fármacos antiepilépticos, foram já documentadas. No entanto, o conhecimento acerca dos efeitos destes fármacos nas células ósseas é ainda escasso. Posto isto, o principal objetivo deste estudo foi investigar o efeito de cinco antiepilépticos diferentes em células ósseas humanas (osteoclastos, osteoblastos e culturas de ambas as células). As culturas celulares de osteoclastos foram instituídas a partir de células percursoras isoladas de sangue periférico humano e caracterizadas para a atividade da TRAP (fosfatase ácida resistente ao tartarato), número de células multinucleadas TRAP positivas, presença de células com anéis de actina e que expressam recetores de vitronectina e calcitonina e taxa de apoptose. Para além disto, também o envolvimento de vias de sinalização na resposta celular foi testado. As culturas celulares de osteoblastos foram obtidas a partir de cabeças de fémur de pacientes (25-45 anos) submetidos a cirurgia ortopédica e foram também caracterizadas para proliferação/viabilidade celular, atividade da ALP (fosfatase alcalina), e taxa de apoptose. Para além disto, também a expressão de genes relacionados com osteoblastos e o envolvimento de vias de sinalização na resposta celular foram estudadas. Relativamente às co-culturas, em primeiro lugar as células osteoblásticas foram semeadas e cultivadas. De seguida, as células mononucleadas do sangue periférico (PBMC) foram adicionadas às células osteoblásticas e as co-culturas foram avaliadas para os mesmos parâmetros mencionados para as células osteoclásticas e osteoblásticas. As culturas celulares estudadas foram mantidas na ausência (controlo) ou na presença de cinco antiepiléticos diferentes (carbamazepina, gabapentina, lamotrigina, topiramato e ácido valpróico). Todos os fármacos testados foram capazes de afetar o desenvolvimento das células osteoclásticas e osteoblásticas, no entanto, mostraram modular de forma diferente estes processos. De um modo geral, a tendência foi para inibir o processo de desenvolvimento de ambas as células. Adicionalmente, as vias de sinalização envolvidas também parecem ter sido afetadas pelos diferentes fármacos, sugerindo que estes podem afetar a osteoclastogénese e a osteoblastogénese por diferentes mecanismos. Em jeito de conclusão, o presente estudo mostrou que os diferentes fármacos antiepilépticos possuem a capacidade de modular direta e indiretamente a diferenciação das células ósseas, fornecendo novas luzes para uma melhor compreensão de como estes fármacos podem afetar o tecido ósseo.Instituto Politécnico do Porto. Escola Superior de Tecnologia da Saúde do PortoRodrigues, JoãoPrudêncio, CristinaFerraz, RicardoRepositório Científico do Instituto Politécnico do PortoRocha, Sara2014-01-27T11:45:28Z20132013-01-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttp://hdl.handle.net/10400.22/3470TID:201167611enginfo:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2023-03-13T12:43:17Zoai:recipp.ipp.pt:10400.22/3470Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T17:24:27.078486Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse
dc.title.none.fl_str_mv Characterization of the effects of antiepileptic drugs on bone metabolism: in vitro studies with human osteoblastic and osteoclastic cells
title Characterization of the effects of antiepileptic drugs on bone metabolism: in vitro studies with human osteoblastic and osteoclastic cells
spellingShingle Characterization of the effects of antiepileptic drugs on bone metabolism: in vitro studies with human osteoblastic and osteoclastic cells
Rocha, Sara
Bone remodeling
Osteoclastic cells
Osteoblastic cells
Osteoclastogenesis
Osteoblastogenesis
Antiepileptic drugs
Epilepsy
Remodelação óssea
Osteoclastos
Osteoblastos
Osteoclastogénese
Osteoblastogénese
Antiepilépticos
Epilepsia
title_short Characterization of the effects of antiepileptic drugs on bone metabolism: in vitro studies with human osteoblastic and osteoclastic cells
title_full Characterization of the effects of antiepileptic drugs on bone metabolism: in vitro studies with human osteoblastic and osteoclastic cells
title_fullStr Characterization of the effects of antiepileptic drugs on bone metabolism: in vitro studies with human osteoblastic and osteoclastic cells
title_full_unstemmed Characterization of the effects of antiepileptic drugs on bone metabolism: in vitro studies with human osteoblastic and osteoclastic cells
title_sort Characterization of the effects of antiepileptic drugs on bone metabolism: in vitro studies with human osteoblastic and osteoclastic cells
author Rocha, Sara
author_facet Rocha, Sara
author_role author
dc.contributor.none.fl_str_mv Rodrigues, João
Prudêncio, Cristina
Ferraz, Ricardo
Repositório Científico do Instituto Politécnico do Porto
dc.contributor.author.fl_str_mv Rocha, Sara
dc.subject.por.fl_str_mv Bone remodeling
Osteoclastic cells
Osteoblastic cells
Osteoclastogenesis
Osteoblastogenesis
Antiepileptic drugs
Epilepsy
Remodelação óssea
Osteoclastos
Osteoblastos
Osteoclastogénese
Osteoblastogénese
Antiepilépticos
Epilepsia
topic Bone remodeling
Osteoclastic cells
Osteoblastic cells
Osteoclastogenesis
Osteoblastogenesis
Antiepileptic drugs
Epilepsy
Remodelação óssea
Osteoclastos
Osteoblastos
Osteoclastogénese
Osteoblastogénese
Antiepilépticos
Epilepsia
description Bone is constantly being molded and shaped by the action of osteoclasts and osteoblasts. A proper equilibrium between both cell types metabolic activities is required to ensure an adequate skeletal tissue structure, and it involves resorption of old bone and formation of new bone tissue. It is reported that treatment with antiepileptic drugs (AEDs) can elicit alterations in skeletal structure, in particular in bone mineral density. Nevertheless, the knowledge regarding the effects of AEDs on bone cells are still scarce. In this context, the aim of this study was to investigate the effects of five different AEDs on human osteoclastic, osteoblastic and co-cultured cells. Osteoclastic cell cultures were established from precursor cells isolated from human peripheral blood and were characterized for tartrate-resistant acid phosphatase (TRAP) activity, number of TRAP+ multinucleated cells, presence of cells with actin rings and expressing vitronectin and calcitonin receptors and apoptosis rate. Also, the involvement of several signaling pathways on the cellular response was addressed. Osteoblastic cell cultures were obtained from femur heads of patients (25-45 years old) undergoing orthopaedic surgery procedures and were then studied for cellular proliferation/viability, ALP activity, histochemical staining of ALP and apoptosis rate. Also the expression of osteoblast-related genes and the involvement of some osteoblastogenesis-related signalling pathways on cellular response were addressed. For co-cultured cells, osteoblastic cells were firstly seeded and cultured. After that, PBMC were added to the osteoblastic cells and co-cultures were evaluated using the same osteoclast and osteoblast parameters mentioned above for the corresponding isolated cell. Cell-cultures were maintained in the absence (control) or in the presence of different AEDs (carbamazepine, gabapentin, lamotrigine, topiramate and valproic acid). All the tested drugs were able to affect osteoclastic and osteoblastic cells development, although with different profiles on their osteoclastogenic and osteoblastogenic modulation properties. Globally, the tendency was to inhibit the process. Furthermore, the signaling pathways involved in the process also seemed to be differently affected by the AEDs, suggesting that the different drugs may affect osteoclastogenesis and/or osteoblastogenesis through different mechanisms. In conclusion, the present study showed that the different AEDs had the ability to directly and indirectly modulate bone cells differentiation, shedding new light towards a better understanding of how these drugs can affect bone tissue.
publishDate 2013
dc.date.none.fl_str_mv 2013
2013-01-01T00:00:00Z
2014-01-27T11:45:28Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/masterThesis
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dc.identifier.uri.fl_str_mv http://hdl.handle.net/10400.22/3470
TID:201167611
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dc.publisher.none.fl_str_mv Instituto Politécnico do Porto. Escola Superior de Tecnologia da Saúde do Porto
publisher.none.fl_str_mv Instituto Politécnico do Porto. Escola Superior de Tecnologia da Saúde do Porto
dc.source.none.fl_str_mv reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
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