Restor Dent Endod.  2012 Mar;37(1):34-40. 10.5395/rde.2012.37.1.34.

Morphological evaluation during in vitro chondrogenesis of dental pulp stromal cells

• Chung CR
• Kim HN
• Park Y
• Kim MJ
• Oh YJ
• Shin SJ
• Choi YJ
• Kim KH

• KMID: 1995682
• DOI: http://doi.org/10.5395/rde.2012.37.1.34

Abstract

OBJECTIVES
The aim was to confirm the stem cell-like properties of the dental pulp stromal cells and to evaluate the morphologic changes during in vitro chondrogenesis.
MATERIALS AND METHODS
Stromal cells were outgrown from the dental pulp tissue of the premolars. Surface markers were investigated and cell proliferation rate was compared to other mesenchymal stem cells. Multipotency of the pulp cells was confirmed by inducing osteogenesis, adipogenesis and chondrogenesis. The morphologic changes in the chondrogenic pellet during the 21 day of induction were evaluated under light microscope and transmission electron microscope. TUNEL assay was used to evaluate apoptosis within the chondrogenic pellets.
RESULTS
Pulp cells were CD90, 105 positive and CD31, 34 negative. They showed similar proliferation rate to other stem cells. Pulp cells differentiated to osteogenic, adipogenic and chondrogenic tissues. During chondrogenesis, 3-dimensional pellet was created with multi-layers, hypertrophic chondrocyte-like cells and cartilage-like extracellular matrix. However, cell morphology became irregular and apoptotic cells were increased after 7 day of chondrogenic induction.
CONCLUSIONS
Pulp cells indicated mesenchymal stem cell-like characteristics. During the in vitro chondrogenesis, cellular activity was superior during the earlier phase (within 7 day) of differentiation.

Keyword

Cartilage; Chondrogenic pellet; Dental pulp stem cell; Dental pulp stromal cell; in vitro chondrogenesis

MeSH Terms

Adipogenesis
Apoptosis
Bicuspid
Cartilage
Cell Proliferation
Chondrogenesis
Dental Pulp
Durapatite
Electrons
Extracellular Matrix
In Situ Nick-End Labeling
Light
Mesenchymal Stromal Cells
Osteogenesis
Stem Cells
Stromal Cells
Durapatite

Figure

• 

  Figure 1 Characterization of the dental pulp stromal cells. (a) Fibroblast-like cells were collected from dental pulp tissue; (b) Pulp cells were positive for mesenchymal cell surface markers CD 90 and 105; (c) Proliferation rates of pulp cells from premolar (DP4), and 3rd molar (DP8) were similar to those of bone marrow mesenchymal stem cell (BMMSC) and adipogenic stem cell (ADSC).

• 

  Figure 2 Multipotency of the pulp stromal cells. (a) Osteogenesis; (b) Adiopogenesis; (c) Chondrogenesis after 14 days of induction. Scale bar = 50 mm

• 

  Figure 3 Morphologic evaluation of the chondrogenic pellets engineered from pulp cells. HE (a, b, c, d) and Alcian blue (e, f, g, h) staining at 3, 7, 14 and 21 days of chondrogenic induction. Scale bar = 50 µm. TEM (i, j, k, l) images of the cells within the pellets at 3, 7, 14 and 21 days of chondrogenic induction. White arrows, nucleolus; black arrow, extracellular matrix fibers, scale bar = 5 µm. HE, Hematoxylin-eosin; TEM, transmission eletron microscope.

• 

  Figure 4 Evaluation of apoptotic cells within the chondrogenic pellets. (a) At day 3, few Tunel positive cells were localized only within the superficial layer, but as induction period increased Tunel positive cells were scattered throughout the pellet; (b) at day 7; (c) at day 14; (d) at day 21. Scale bar = 50 µm

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