• JoomlaWorks Simple Image Rotator
  • JoomlaWorks Simple Image Rotator
  • JoomlaWorks Simple Image Rotator
  • JoomlaWorks Simple Image Rotator
  • JoomlaWorks Simple Image Rotator
  • JoomlaWorks Simple Image Rotator
  • JoomlaWorks Simple Image Rotator
  • JoomlaWorks Simple Image Rotator
  • JoomlaWorks Simple Image Rotator
  • JoomlaWorks Simple Image Rotator
 
  Bookmark and Share
 
 
Doctoral Thesis
DOI
Document
Author
Full name
Monica Yuri Orita Misawa
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2018
Supervisor
Committee
Villar, Cristina Cunha (President)
Aranha, Ana Cecilia Corrêa
Araújo, Mauricio Guimarães
Saraiva, Luciana
Title in Portuguese
Caracterização do efeito imunomodulatório das células mesenquimais indiferenciadas na resposta inflamatória a Porphyromonas gingivalis
Keywords in Portuguese
células tronco
inflamação
neutrófilos
Porphyromonas gingivalis
Abstract in Portuguese
O objetivo deste estudo foi caracterizar as respostas das células mesenquimais indiferenciadas derivadas do ligamento periodontal (PDLSCs) ao extrato proteico total de Porphyromonas gingivalis (PgPE) e avaliar seu impacto nas propriedades biológicas das células leucêmicas promielocíticas humanas HL-60. PDLSCs enriquecidas com CD105 foram semeadas em placas de 6 poços durante 24h. Em seguida, as células foram desafiadas com PgPE (0 e 2 mg/ml) durante 3h (período de exposição). Os sobrenadantes foram então descartados; PDLSCs foram lavadas com PBS e cultivadas por 18h adicionais. Por fim, os sobrenadantes foram coletados. Os níveis de citocinas e quimiocinas nos sobrenadantes foram avaliados por ensaios multiplex. Na sequência, o efeito dos sobrenadantes derivados de PDLSCs (tratadas ou não com PgPE) sobre a ativação, o recrutamento e a resposta inflamatória de HL-60 foi avaliado. PDLSCs responderam ao tratamento com PgPE. RANTES, eotaxina e IP-10 (proteína produzida por IFN-?) foram detectados apenas em sobrenadantes de PDLSCs/PgPE. Além disso, PgPE induziu maior secreção de proteína quimiotática de neutrófilos (MCP)-1, intérferon (IFN)-?, interleucina (IL)-6, IL-8 e IL-1ra (p> 0,05). O recrutamento de HL-60D aumentou 4,7 vezes quando estas células foram expostas a sobrenadantes PDLSCs/PgPE, enquanto que os sobrenadantes de PDLSCs não afetaram a quimiotaxia de HL-60D. Sobrenadantes PDLSCs promoveram uma redução de 16% na produção de espécies de oxigênio reativo (ROS) por HL-60D estimuladas por PMA (p=0,013). Em contraste, sobrenadantes PDLSCs/PgPE promoveram um aumento de 1,78±1,04 vezes (p=0,046) na produção de ROS. Finalmente, tanto sobrenadantes PDLSCs, como sobrenadantes PDLSCs/PgPE, não influenciaram a produção de fator de necrose tumoral (TNF)-? e IL-1? pelas HL-60D em resposta ao lipopolissacarídeo (LPS). Esses achados sugerem um importante papel das PDLSCs no reconhecimento de P. gingivalis, recrutamento de células imunes inatas e ativação de mecanismos antimicrobianos.
Title in English
Role of periodontal-derived mesenchymal stem cells in Porphyromonas gingivalis infection
Keywords in English
inflammation
neutrophils
Porphyromonas gingivalis
stem cells
Abstract in English
The aim of this study was to characterize periodontal ligament-derived mesenchymal stem cells (PDLSCs) responses to Porphyromonas gingivalis total protein extract (PgPE) and its impact on human leucocyte promyelocyte cells HL-60 biological properties. CD105-enriched PDLSCs were seeded in 6-well plates for 24h. Next, cells were challenged with PgPE (0 and 2mg/ml) for 3h (exposure period). Supernatants were then discarded, cells were washed with PBS, and cultured further for 18h before supernatants were collected. Supernatants' cytokine and chemokine levels were assessed by bead-based multiplex assays. The effect of supernatants collected from untreated and PgPE-treated PDLSCs on HL-60D activation, recruitment and inflammatory responses was determined. PDLSCs were responsive to PgPE treatment. RANTES, eotaxin, and interferon-inducible protein (IP)-10 were detected only in supernatants collected from PgPE treated cells. Moreover, PgPE induced higher monocyte chemoattractant protein (MCP)-1, interferon (IFN)-?, interleukin (IL)-6, IL-8 and IL-1ra secretion (p > 0.05). HL-60D recruitment was increased by 4.7 fold when exposed to PDLSCs/PgPE supernatants, whereas PDLSCs supernatants did not affect HL-60D chemotaxis. PDLSCs supernatants promoted a 16% reduction in radical oxygen species (ROS) production by PMA-stimulated HL-60D (p=0.013). On the contrary, PDLSCs/PgPE supernatants promoted a 1.78 ± 1.04 fold increase (p=0.046) in ROS production. Finally, both PDLSCs and PDLSCs/PgPE supernatants had no effect on HL-60D tumor necrosis factor (TNF)-? and IL-1? responses to lipopolysaccharide (LPS). These findings strongly suggest an important role of PDLSCs in the recognition of P gingivalis, recruitment of innate immune cells and activation of antimicrobial mechanisms.
 
WARNING - Viewing this document is conditioned on your acceptance of the following terms of use:
This document is only for private use for research and teaching activities. Reproduction for commercial use is forbidden. This rights cover the whole data about this document as well as its contents. Any uses or copies of this document in whole or in part must include the author's name.
Publishing Date
2019-05-28
 
WARNING: Learn what derived works are clicking here.
All rights of the thesis/dissertation are from the authors
CeTI-SC/STI
Digital Library of Theses and Dissertations of USP. Copyright © 2001-2020. All rights reserved.