J Korean Acad Periodontol.  2009 Jun;39(2):119-128. 10.5051/jkape.2009.39.2.119.

Investigation of postnatal stem cells from canine dental tissue and bone marrow

• Jhin MJ
• Kim YS
• Kim SH
• Kim KH
• Lee CW
• Koo KT
• Kim TI
• Seol YJ
• Ku Y
• Rhyu IC
• Chung CP
• Lee YM

• KMID: 1733729
• DOI: http://doi.org/10.5051/jkape.2009.39.2.119

Abstract

PURPOSE: The aim of this study was to evaluate the stemness of cells from canine dental tissues and bone marrow.
METHODS
Canine periodontal ligament stem cells (PDLSC), alveolar bone stem cells (ABSC) and bone marrow stem cells(BMSC) were isolated and cultured. Cell differentiations (osteogenic, adipogenic and chondrogenic) and surface antigens (CD146, STRO-1, CD44, CD90, CD45, CD34) were evaluated in vitro. The cells were transplanted into the subcutaneous space of nude mice to assess capacity for ectopic bone formation at 8 weeks after implantation.
RESULTS
PDLSC, ABSC and BMSC differentiated into osteoblasts, adipocytes and chondrocytes under defined condition. The cells expressed the mesenchymal stem cell markers differently. When transplanted into athymic nude mice, these three kinds of cells with hydroxyapatite /beta tricalcium phosphate (HA/TCP) carrier showed ectopic bone formation.
CONCLUSIONS
This study demonstrated that canine dental stem cells have stemness like bone marrow stem cells. Transplantation of these cells might be used as a therapeutic approach for dental stem cell-mediated periodontal tissue regeneration.

Keyword

stem cells; differentiation; periodontal ligament; regeneration

MeSH Terms

Adipocytes
Animals
Antigens, Surface
Bone Marrow
Calcium Phosphates
Cell Differentiation
Chondrocytes
Durapatite
Mesenchymal Stromal Cells
Mice
Mice, Nude
Osteoblasts
Osteogenesis
Periodontal Ligament
Regeneration
Stem Cells
Transplants
Antigens, Surface
Calcium Phosphates
Durapatite

Figure

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  Figure 1 The cultured canine PDLSCs form adherent clonogenic cell clusters of fibroblast-like cells (A) Colony forming Unit (Original magnification ×40) (B) typical fibroblast-like cells (Original magnification ×100).

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  Figure 2 Osteogenic differentiation (A) PDLSC (B) ABSC (C) BMSC (Alizarin red S, Original magnification ×100).

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  Figure 3 Adipogenic differentiation (A) PDLSC (B) ABSC (C) BMSC (Oil Red O staining, Original magnification ×100).

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  Figure 4 Chodrogenic differentiation (A) PDLSC (B) ABSC (C) BMSC (Alcian Blue staining, Original magnification ×200).

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  Figure 5 Flow cytometric analysis of PDLSC(A) 12.96% of PDLSC were positive staining for CD146 (B) 99.76% for CD44 (C) 99.40% for CD90 (D) 0.25% for STRO-1 (E) 5.85% for CD45 (F) 5.57% for CD34.

• 

  Figure 6 Flow cytometric analysis of ABSC (A) 38.97% of ABSC were positive staining for CD146 (B) 99.38% for CD44 (C) 99.24% for CD90 (D) 0.23% for STRO-1 (E) 9.56% for CD45 (F) 7.20% for CD34.

• 

  Figure 7 Flow cytometric analysis of BMSC (A) 11.49% of BMSC were positive staining for CD146 (B) 99.50% for CD44 (C) 86.39% for CD90 (D) 3.02% for STRO-1 (E) 3.47% for CD45 (F) 2.88% for CD34.

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  Figure 8 BMSCs attached to HA/TCP. The arrow indicates attached cells (SEM, Original magnification ×100, ×300).

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  Figure 9 Eight weeks after transplantation. The asterisk(*) indicates new bone formation aroundHA/TCP scaffolds (A) PDLSC (B) ABSC (C) BMSC (D) Control (Multiple staining, Original magnification ×200).

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