Korean J Orthod.  2012 Oct;42(5):249-254. 10.4041/kjod.2012.42.5.249.

Skeletal myogenic differentiation of human periodontal ligament stromal cells isolated from orthodontically extracted premolars

• Song M
• Kim H
• Choi Y
• Kim K
• Chung C

• KMID: 1435400
• DOI: http://doi.org/10.4041/kjod.2012.42.5.249

Abstract

OBJECTIVE
To investigate the stem cell-like characteristics of human periodontal ligament (PDL) stromal cells outgrown from orthodontically extracted premolars and to evaluate the potential for myogenic differentiation.
METHODS
PDL stromal cells were obtained from extracted premolars by using the outgrowth method. Cell morphological features, self-replication capability, and the presence of cell-surface markers, along with osteogenic, adipogenic, and chondrogenic differentiation, were confirmed. In addition, myogenic differentiation was induced by the use of 5-aza-2'-deoxycytidine (5-Aza) for DNA demethylation.
RESULTS
PDL stromal cells showed growth patterns and morphological features similar to those of fibroblasts. In contrast, the proliferation rates of premolar PDL stromal cells were similar to those of bone marrow and adipogenic stem cells. PDL stromal cells expressed surface markers of human mesenchymal stem cells (i.e., CD90 and CD105), but not those of hematopoietic stem cells (i.e., CD31 and CD34). PDL stromal cells were differentiated into osteogenic, adipogenic, and chondrogenic lineages. Myotube structures were induced in PDL stromal cells after 5-Aza pretreatment, but not in the absence of 5-Aza pretreatment.
CONCLUSIONS
PDL stromal cells isolated from extracted premolars can potentially be a good source of postnatal stem cells for oromaxillofacial regeneration in bone and muscle.

Keyword

Human periodontal ligament stem cell; Myogenic differentiation; Cell biology; Molecular biology; Soft tissue

MeSH Terms

Azacitidine
Bicuspid
Bone Marrow
DNA
Durapatite
Fibroblasts
Hematopoietic Stem Cells
Humans
Mesenchymal Stromal Cells
Molecular Biology
Muscle Fibers, Skeletal
Muscles
Periodontal Ligament
Regeneration
Stem Cells
Stromal Cells
Azacitidine
DNA
Durapatite

Figure

• 

  Figure 1 Proliferation of periodontal ligament (PDL) stromal cells compared to that of other cell types. A, The morphological features of PDL stromal cells were similar to those of spindle-like fibroblasts (×40). B, Proliferation rates of PDL stromal cells were similar to the rates for ADSCs and BMMCs. ADSC, adipogenic stem cell; BMMC, bone marrow mesenchymal stem cell; HDF, human dermal fibroblast.

• 

  Figure 2 Multipotency of periodontal ligament (PDL) stromal cells. A, PDL stromal cells displayed high expression levels of the mesenchymal stem cell surface markers CD90 and CD105, but very low expression levels of the hematopoietic stem cell surface markers CD31 and CD34. Gray solid fill, isotype; black line, marker of interest. B, Calcified nodules, lipid cluster formation, and chondrogenic pellet formation were noted after osteogenic, adipogenic, and chondrogenic induction, respectively.

• 

  Figure 3 Myotube formation and desmin expression of periodontal ligament (PDL) stromal cells. Control cells cultured in the absence of 5-aza-2'-deoxycytidine (5-Aza) treatment, A and cells pretreated with 5-Aza for DNA demethylation, B. Myotube-like structures that were positive for desmin (B, green) were only observed in B. Cell nucleus counterstained with DAPI (blue).

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