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Previous issue date: 2015-06-15 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / Teeth exhibit limited repair in response to damage, and studies have shown that dental
pulp stem cells (DPSCs) probably provide a source of cells to replace those damaged and
to facilitate repair. Additionally, in vivo transplantation into immunocompromised mice
demonstrated the ability of DPSCs to generate functional dental tissue in the form of
dentine/pulp-like complexes. Therefore, the purpose of this study was to characterize the
endothelial fate of DPSCs in vivo and to start to understand the role of Tie2/Ang-1
signaling pathway on the regulation of the endothelial differentiation of DSPCs.! DPSCs
stably transduced with LacZ were seeded into tooth slices and implanted into
immunodeficient mice for 7, 14, 21 ad 28 days. A combination of in vitro and in vivo
assays were performed to determine the role of Ang-1 in the vasculogenic fate of DPSCs.
Histologic analysis and immunohistochemistry from the retrieved tooth slices were
performed. Dental pulp stem cells seeded in tooth slice/scaffolds and transplanted into
SCID mice showed a continuous crescent number of beta-galactosidase-positive
odontoblastic and endothelial cells along the 7, 14, 21 and 28 days of the study. At 21
days beta-galactosidase-positive capillaries located close to murine blood vessels were
detected, confirming our previous report (Cordeiro et al, 2008; Sakai et al, 2010). DPSCs
exposed to Ang-1 differentiated into cells expressing VEFGR2, CD31 and Tie2.
Exposure of VEGFR1-silenced DSPC to Ang-1 and VEGF induced the activation of
STAT3, Erk and Akt, while VEGF alone inhibited the phosphorylation of STAT3. This is
consistent with the role of Akt and STAT3 in the regulation of cell survival and cell-cell
interation of DPSC through Tie2/Ang-1 signaling pathway. This study demonstrates that
Angiopoietin-1 is involved in the vasculogenic differentiation of DSPCs. / As c?lulas-tronco provenientes da polpa de dentes adultos (DPSC) s?o uma fonte
promissora de c?lulas-tronco para a terapia regenerativa. J? foi demonstrado que elas
podem se diferenciar em c?lulas endoteliais, mas os relatos histol?gicos temporais do seu
destino ainda n?o foram demonstrados. Dessa forma, o presente estudo teve como
objetivo elucidar o destino das DSPC-LacZ, bem como avaliar o efeito de algumas
mol?culas envolvidas na diferencia??o endotelial destas c?lulas. Para isso, DPSC Lac-Z
foram semeadas em scaffolds de fatias dent?rias e implantadas no subcut?neo de
camundongos imunodeficientes. 14 dias ap?s a recupera??o dos tecidos, observou-se
c?lulas beta-galagtosidase positivas alinhadas ao longo das paredes internas das fatias de
dente, sugerindo a diferencia??o destas em c?lulas semelhantes ? odontoblastos. Ainda
neste per?odo, c?lulas beta-galactosidase positivas foram localizadas pr?ximas aos vasos
sangu?neos do roedor, algumas formaram estruturas tubulares e continham elementos
celulares dentro do seu l?men. Aos 21 dias, um tecido muito semelhante a polpa dental
foi encontrado dentro das fatias dentais, e no dia 28 o n?mero de c?lulas betagalactosidade
postivas era maior do que todos os periodos anteriores. Ainda, avaliou-se o
papel da mol?cula VEGF e Ang-1 na diferencia??o endotelial destas c?lulas bem como
na fosforila??o de algumas vias de sinaliza??o. Para isso, um meio de cultura alpha-
MEM suplementado com 50 ng/mL rhVEGF + 200 ng/mL rhAng-1, foi utlizado como
est?mulo para as DPSC se diferenciarem em c?lulas endoteliais. Durante um per?odo de
28 dias as c?lulas expressaram marcadores endoteliais como VEGFR2, CD31, Ang-1 e
Ang-2. Quando utilizamos c?lulas DPSC shRNA VEGFR-1 estimuladas pelas mesmas
mol?culas, a fosforila??o de ERK e AKT foi aumentada, o que n?o ocorreu no grupo controle.
| Identifer | oai:union.ndltd.org:IBICT/oai:tede2.pucrs.br:tede/7710 |
| Date | 15 June 2015 |
| Creators | Cucco, Carolina |
| Contributors | Figueiredo, Jos? Antonio Poli de, N?r, Jacques Eduardo |
| Publisher | Pontif?cia Universidade Cat?lica do Rio Grande do Sul, Programa de P?s-Gradua??o em Odontologia, PUCRS, Brasil, Faculdade de Odontologia |
| Source Sets | IBICT Brazilian ETDs |
| Language | Portuguese |
| Detected Language | English |
| Type | info:eu-repo/semantics/publishedVersion, info:eu-repo/semantics/doctoralThesis |
| Format | application/pdf |
| Source | reponame:Biblioteca Digital de Teses e Dissertações da PUC_RS, instname:Pontifícia Universidade Católica do Rio Grande do Sul, instacron:PUC_RS |
| Rights | info:eu-repo/semantics/openAccess |
| Relation | -7411869720500764667, 500, 500, 500, 600, 4673435736271820140, -2070498469879244349, 3590462550136975366 |
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