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. 2012;9(6):480-7.
doi: 10.7150/ijms.4583. Epub 2012 Aug 13.

Osteogenic differentiation of dental follicle stem cells

Giorgio Mori  1 , Andrea Ballini, Claudia Carbone, Angela Oranger, Giacomina Brunetti, Adriana Di Benedetto, Biagio Rapone, Stefania Cantore, Mariasevera Di Comite, Silvia Colucci, Maria Grano, Felice R Grassi
Affiliations

Osteogenic differentiation of dental follicle stem cells

Giorgio Mori et al. Int J Med Sci. 2012.

Abstract

Background: Stem cells are defined as clonogenic cells capable of self-renewal and multi-lineage differentiation. A population of these cells has been identified in human Dental Follicle (DF). Dental Follicle Stem Cells (DFSCs) were found in pediatric unerupted wisdom teeth and have been shown to differentiate, under particular conditions, into various cell types of the mesenchymal tissues.

Aim: The aim of this study was to investigate if cells isolated from DF show stem features, differentiate toward osteoblastic phenotype and express osteoblastic markers.

Methods: We studied the immunophenotype of DFSCs by flow cytometric analysis, the osteoblastic markers of differentiated DFSCs were assayed by histochemical methods and real-time PCR.

Results: We demonstrated that DFSCs expressed a heterogeneous assortment of makers associated with stemness. Moreover DFSCs differentiated into osteoblast-like cells, producing mineralized matrix nodules and expressed the typical osteoblastic markers, Alkaline Phosphatase (ALP) and Collagen I (Coll I).

Conclusion: This study suggests that DFSCs may provide a cell source for tissue engineering of bone.

Keywords: bone tissue engineering; dental follicle; human postnatal dental tissue.; osteogenic markers; stem cells.

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Conflict of interest statement

CONFLICT OF INTEREST: The authors have declared that no competing interest exists.

Figures

Figure 1
Figure 1
Phenotypes of DFSCs. The expression of the indicated mesenchymal stem cell markers on DFSCs was analyzed using flow cytometry. Results from one representative DFSC culture are shown. The grey histograms signify staining with isotype controls, and the white histograms represent staining with the specified surface marker antibody.
Figure 2
Figure 2
DFSCs proliferationin vitro. Proliferation of DFSCs and BMSCs after three, five and seven days of culture.
Figure 3
Figure 3
ALP expression. Histochemical staining for ALP in DFSCs incubated in osteogenic medium for 10 days (samples from three different patients) (A). Real Time-PCR analysis shows ALP expression in DFSCs cultured for 10 and 17 days in osteogenic medium: the expression of the enzyme progressively increased up to 3 thousand of times (B).
Figure 4
Figure 4
Mineralized nodules formation. Von Kossa staining shows mineralized nodules formation by DFSCs (from three patients) incubated for thirty days in osteogenic medium.
Figure 5
Figure 5
Coll I and RUNX2 expression. Real Time-PCR analysis shows RUNX2 and Coll I increased expression in DFSCs cultured for 10 and 17 days in osteogenic medium: Coll I increased progressively up to two hundred times and RUNX2 increased up to seven times.
See this image and copyright information in PMC

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