Expression of osteoblastic phenotype in periodontal ligament fibroblasts cultured in three-dimensional collagen gel
J. appl. oral sci
; 23(2): 206-214, Mar-Apr/2015. graf
Article
in English
| LILACS, BBO - Dentistry
| ID: lil-746544
Responsible library:
BR1.1
ABSTRACT
Objective:
To investigate the influence of a three-dimensional cell culture model on the expression of osteoblastic phenotype in human periodontal ligament fibroblast (hPDLF) cultures. Material andMethods:
hPDLF were seeded on bi-dimensional (2D) and three-dimensional (3D) collagen type I (experimental groups) and and on a plastic coverslip (control) for up to 14 days. Cell viability and alkaline phosphatase (ALP) activity were performed. Also, cell morphology and immunolabeling for alkaline phosphatase (ALP) and osteopontin (OPN) were assessed by epifluorescence and confocal microscopy. The expression of osteogenic markers, including alkaline phosphatase, osteopontin, osteocalcin (OC), collagen I (COL I) and runt-related transcription factor 2 (RUNX2), were analyzed using real-time polymerase chain reaction (RT-PCR). Mineralized bone-like nodule formation was visualized by microscopy and calcium content was assessed quantitatively by alizarin red assay.Results:
Experimental cultures produced an increase in cell proliferation. Immunolabeling for OPN and ALP in hPDLF were increased and ALP activity was inhibited by three-dimensional conditions. OPN and RUNX2 gene expression was significantly higher on 3D culture when compared with control surface. Moreover, ALP and COL I gene expression were significantly higher in three-dimensional collagen than in 2D cultures at 7 days. However, at 14 days, 3D cultures exhibited ALP and COL I gene expression significantly lower than the control, and the COL I gene expression was also significantly lower in 3D than in 2D cultures. Significant calcium mineralization was detected and quantified by alizarin red assay, and calcified nodule formation was not affected by tridimensionality.Conclusion:
This study suggests that the 3D cultures are able to support hPDLF proliferation and favor the differentiation and mineralized matrix formation, which may be a potential periodontal regenerative therapy. .
Full text:
Available
Collection:
International databases
Database:
BBO - Dentistry
/
LILACS
Main subject:
Review Literature as Topic
/
Meta-Analysis as Topic
/
Stroke
Type of study:
Prognostic study
/
Systematic review
Limits:
Animals
/
Humans
Language:
English
Journal:
J. appl. oral sci
Journal subject:
Dentistry
Year:
2015
Document type:
Article
Affiliation country:
Brazil
Institution/Affiliation country:
University of São Paulo/BR