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| Author(s): |
Bruno Braga Benatti
Total Authors: 1
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| Document type: | Doctoral Thesis |
| Press: | Campinas, SP. |
| Institution: | Universidade Estadual de Campinas (UNICAMP). Faculdade de Odontologia de Piracicaba |
| Defense date: | 2007-06-27 |
| Examining board members: |
Francisco Humberto Nociti Junior;
Giuseppe Alexandre Romito;
Poliana Mendes Duarte;
Reginaldo Bruno Gonçalves;
Karina Gonzales Silverio
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| Advisor: | Francisco Humberto Nociti Junior |
| Abstract | |
The aim of this study was to investigate the effect of aging on periodontal ligament cells (PDLC), modulated by the treatment with bFGF. Primary periodontal ligament cell cultures were obtained and divided into the following experimental groups: A ¿ impacted (cells from impacted molars from subjects aged 18 to 30 years), B ¿ young (cells from erupted pre-molars from subjects aged 15 to 20 years), C ¿ aged (cells from erupted teeth from patients aged more than 60 years), D ¿ group A treated with 10?g/ml of bFGF, E ¿ group B treated with 10?g/ml of bFGF and F ¿ group C treated with 10?g/ml of bFGF. For all groups, we performed proliferation and cell viability assays, mineralization assay, total protein quantification and assessed mRNA levels of: type I and III collagen, MMP-2 e -8, TIMP-1 e ¿2, PDGF-1, BMP-3, bFGF, OPG, RANKL, IL-1, IL-4, IL-6, IL-8 and FGFR 1 utlizing the ¿real-time¿ PCR technique. Data analysis demonstrated that aging negatively influenced cell proliferation and mineral nodule formation (P<0.05), while cell viability and total protein secretion were not affected (P>0.05). Gene expression analysis revealed that aging decreased mRNA levels of collagen type I and IL-4, and increased mRNA levels of FGFR1, MMP-8, TIMP-1, OPG and IL-1 and 6 (P<0.05). Treatment with 10?g/ml of bFGF significantly increased cell proliferation for all groups (P<0.05), however with a less impact on aged cells (P<0.05). In addition, cell viability and total protein secretion were not affected by bFGF (P>0.05), while mineral nodule formation was significantly inhibited (P<0.05). Finally, gene expression analysis demonstrated that bFGF decreased mRNA levels of all genes (P<0.05), except for TIMP-1 and IL-8 that were increased (P<0.05), PDGF-1 that was not affected (P>0.05) and BMP-3 that was increased in young cells and decreased in aged cells (P<0.05). Within the limits of our study, we may conclude that: i) it was possible to esblish primary cultures of PDLC obtained from young and aged donors, ii) aging modulates important features of periodontal ligament cells (PDLC), decreasing the potential for regeneration of mineralized tissues and favoring a pro-inflammatory and degenerative profile and iii) treatment with 10?g/ml of bFGF leads to similar PDLC behavioral alterations despite of the donor age (AU) | |