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Doctoral Thesis
DOI
https://doi.org/10.11606/T.23.2016.tde-19012016-165347
Document
Author
Full name
Cleber Keiti Nabeshima
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2015
Supervisor
Committee
Machado, Manoel Eduardo de Lima (President)
Chavez, Victor Elias Arana
Duarte, Marco Antonio Hungaro
Gavini, Giulio
Souza, Arlindo di Spagna
Title in Portuguese
Análise do efeito do fator de crescimento endotelial na angiogênese em canais de dentes de ratos com apicigênese completa
Keywords in Portuguese
Angiogênese
Apicigênese completa
Fator de crescimento endotelial
Abstract in Portuguese
A bioengenharia tecidual tem sido aplicada na Endodontia na busca de tratamentos mais biológicos. A revascularização pulpar tem sido observada em dentes com ápice aberto, porém há poucos estudos direcionados aos dentes com ápice formado. A angiogênese é primordial para a nutrição celular durante a regeneração tecidual. O objetivo deste estudo foi analisar o efeito do fator de crescimento endotelial (VEGF) na angiogênese após pulpectomia de canais radiculares de rato com apicigênese completa. A câmara pulpar dos primeiros molares superiores de 12 ratos machos Wistar com 13 semanas foi acessada. A polpa radicular do canal mésio vestibular foi removida e o canal instrumentado até a lima K25. Com uma lima K20, além do limite foraminal, foi induzido o sangramento periapical para o canal radicular. Os dentes do lado direito da arcada foram preenchidos com coágulo sanguíneo (GS), e nos dentes do lado esquerdo foi utilizado VEGF165 recombinante (Prospec, Israel) adicionado ao coágulo (GV). Os dentes foram selados com ionômero de vidro fotopolimerizável, e 60 dias depois os animais foram sacrificados. A maxila foi dissecada, fixada, descalcificada e incluída em parafina. Cortes transversais seriados foram feitos e corados com hematoxilina-eosina para avaliação morfológica ou incubados com anticorpo de coelho anti-fator VIII (Bioss, EUA) para análise da angiogênese. A análise histológica demonstrou canais preenchidos por tecido conjuntivo sem presença de odontoblastos nos dois grupos. A imuno-histoquímica demonstrou positividade para o anticorpo no tecido neoformado em ambos os grupos. No grupo 1 foi predominante a marcação difusa, e no grupo 2 foi a marcação definida. Pode-se concluir que o uso de VEGF acelera a angiogênese que ocorre em canais pulpectomizados de molares com apicigênese completa.
Title in English
Analysis of the effect of vascular endothelial growth factor on angiogenesis of root canal in mature teeth of rats
Keywords in English
Angiogenesis
Mature teeth
Vascular endothelial growth factor
Abstract in English
The tissue engineering has been applied in Endodontics in search of more biological treatments. The pulp revascularization has been observed in immature teeth, but there are few studies directed to the closed-apex teeth. Angiogenesis is essential for cellular nutrition during tissue regeneration. The aim of this study was to analyze the effect of vascular endothelial growth factor (VEGF) in angiogenesis after pulp removal in rats with mature teeth. The pulp chamber of upper first molars of 12 male Wistar rats with 13-weeks-old was accessed. The pulp of the mesiobuccal root canal was removed, and the root canal was shaped up to 25K-file. Then, a 20K-file was introduced beyond the apex to induce bleeding. The teeth on the right side of the arch are filled using blood clot alone (GS) and the teeth on the left side was filled using blood clot + VEGF165 recombinant (Prospec, Israel) (GV). The teeth were coronally sealed using light-curing glass ionomer, and the animals were sacrificed after 60 days. The samples were dissected, fixed, decalcified and included in paraffin. Transverse serial sections were made, and stained using hematoxylin-eosin to morphological evaluation or expressed by immunostaining using anti-Factor VIII rabbit antibody (BIOSs, USA) to angiogenesis analysis. Morphological analysis showed root canals filled by connective tissue without the presence of odontoblasts in both groups. Immunohistochemistry showed positivity for the antibody in the new tissue in both groups, with marking defined in group 2. Diffuse marking was predominant in the group 1, and the well-maked type was predominant in the group 2. It can be concluded that the use of VEGF accelerates angiogenesis that occurs in root canal of closed-apex molars after pulpectomy.
 
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Publishing Date
2016-02-12
 
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