Dissertação (mestrado)—Universidade de Brasília, Faculdade de Ciências da Saúde, 2008. / Submitted by Danyelle Mayara Silva (danielemaiara@gmail.com) on 2009-09-14T20:28:03Z
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Previous issue date: 2008 / Células-tronco adultas representam uma nova abordagem em terapia
regenerativa. Recentes pesquisas revelaram que o tecido pulpar de dentes humanos
permanentes e decíduos contém células-tronco com grande potencial proliferativo,
apresentam capacidade de auto-renovação e de diferenciação em diversas linhagens
celulares in vitro. O objetivo deste projeto foi comparar o perfil morfológico e
proliferativo dos tipos celulares cultivados e caracterizar as células-tronco pulpares
humanas de dentes permanentes (CPdp) e decíduos (CPdd) em relação a dois métodos
de isolamento. Terceiros molares impactados e dentes decíduos recém-esfoliados
foram coletados, limpos e seccionados no limite da junção cemento-esmalte. Os
tecidos pulpares foram cuidadosamente removidos e cultivados, utilizando os métodos
de isolamento por digestão enzimática com solução de 3mg/mL de colagenase tipo I e
4mg/mL de dispase e pela cultura direta do fragmento do tecido pulpar. Realizamos
análises morfológicas e do potencial proliferativo. Para a caracterização do perfil
imunofenótipo foram utilizados anticorpos monoclonais anti: CD117, CD34, CD45
RA e a avaliação em citometria de fluxo. Nossos resultados indicaram grande
potencial proliferativo in vitro de CPdp e CPdd, principalmente para CPdd. Os tipos
celulares apresentaram imunofenótipo compatível para células-tronco, sendo
expressivamente positivos para CD117, independente do método de isolamento.
Houve diferença na expressão do CD34 entre os grupos celulares. A análise fenotípica
final demonstrou as CPdp como CD117+, CD34- e CD45-, e as CPdd como CD117+,
CD34+ e CD45-.
_____________________________________________________________________________________ ABSTRACT / Adult Stem cells represents a new approach in regenerative therapies.
Recents researches have shown that postnatal and deciduous dental pulp contains stem
cells with high proliferation capacity, ability to self-renew and to differentiate into
multiple cell lineages. The aim of this study was to compare the morphological and
proliferative profiles of the cells types in dental pulp of permanents and deciduous
teeth, and to characterize the progeny by two isolation methods in vitro. Normal
human impacted third molars and exfoliated deciduous teeth were collected, cleaned
and cut around the cementum-enamel junction. The pulp tissue was gently separated
from de crown and root. Pulp cell cultures were established via two approaches by
enzyme digestion in a solution of 3mg/mL collagenase type I and 4mg/mL dispase
and by culture of the explants into a tissue culture dishes. We performed
morphological, proliferative analyses and immunophenotype studies to characterize
the progeny using a cytometer for monoclonal antibodies: CD117, CD34, CD45 RA.
Our findings indicated high proliferative rates for permanents and deciduous teeth,
mainly for deciduous teeth. The cells population showed phenotypes compatibles for
stem cells, with remarkable positive expression of the marker CD117, irrespective of
the isolation method. We identified significant difference for the expression of CD34
among the cells groups. The immunophenotype profiles were CD117+, CD34- and
CD45- for cells from permanents teeth, and CD117+, CD34+ and CD45- for cells from
deciduous teeth.
| Identifer | oai:union.ndltd.org:IBICT/oai:repositorio.unb.br:10482/1683 |
| Date | January 2008 |
| Creators | Souza, Leliane Macedo de |
| Contributors | Toledo, Orlando Ayrton de, Brígido, Marcelo de Macedo |
| Source Sets | IBICT Brazilian ETDs |
| Language | Portuguese |
| Detected Language | English |
| Type | info:eu-repo/semantics/publishedVersion, info:eu-repo/semantics/masterThesis |
| Source | reponame:Repositório Institucional da UnB, instname:Universidade de Brasília, instacron:UNB |
| Rights | info:eu-repo/semantics/openAccess |
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