Chinese Medical Sciences Journal ›› 2017, Vol. 32 ›› Issue (2): 83-91.doi: 10.24920/J1001-9294.2017.012

• Original Article • Previous Articles     Next Articles

Effects of Aging on the Proliferation and Differentiation Capacity of Human Periodontal Ligament Stem Cells

Du Tingting1, †, Liu Na1, †, Gu Bin1, Li Ying1, Yuan Yifang1, Zhang Wei2, Zhang Tong1, *()   

  1. 1 Department of Stomatology, Chinese PLA General Hospital, Beijing 100853, China
    2 Technical Institute of Physics and Chemistry, Beijing 100190, China
  • Received:2017-01-22 Online:2017-06-20 Published:2017-06-10
  • Contact: Zhang Tong E-mail:kqzhengji301@163.com
  • Supported by:
    △Supported by National Natural Science Foundation of China(51473175), Science and Technology Nova Plan of Beijing City(Z141107001814101).

Abstract:

Objective The aim of this study is to investigate the proliferation, differentiation and apoptosis of periodontal ligament stem cells (PDLSC) derived from different aged donors, and to evaluate the effects of aging on the biological characteristics of PDLSC.Methods Periodontal ligament tissues were obtained from 24 surgically extracted human premolars during orthodontics therapy. The specimens were divided into three groups according to the donor’s age. Group A: 18-20 years, group B: 30-35 years, group C: 45-50 years. PDLSC were isolated and cultured using a tissue-block-based enzymolytic method by limiting dilution assay. The colony forming efficiency of PDLSC for three experimental groups was determined. Senescence-Associated β-Galactosidase (SA-β-G) expression in the three groups was examined using β-galactosidase staining working solution. Cell cycle and apoptosis of the PDLSC were examined by the flow cytometry. Alkaline phosphatase (ALP) activity was evaluated by ALP staining. The expression of osteoplastic differentiation related genes Runt-related transcription factor-2 (Runx-2), Collagen Type 1 (col-1), and ALP of PDLSC were examined by quantitative real-time RT-PCR.Results The colony forming efficiency of PDLSC in Group A, B and C was 36.67%, 22.67% and 9.33%, respectively, which decreased with donors’ age (P<0.05). SA-β-G expression of the senescent PDLSC in group A, B and C were 4.14%, 16.39%, 50.38%, respectively (P<0.05). Cells in G2/S phase was 38.73%, 29.88%, 18.25% (P<0.05), and the apoptosis rate was 1.57%, 4.56%, 5.84% (P<0.05), in group A, B and C respectively. The ALP staining in the three groups decreased with the increase of donors’ ages, and the expression of Runx-2, col-1 and ALP decreased gradually from group A to group C (all P<0.05), which indicated the osteogenic differentiation capacity of PDLSC decreased while donor aging.Conclusion Human PDLSC could be successfully isolated from periodontal ligament tissues of different aged donors. However, the proliferation and osteogenic differentiation capacity of PDLSC decreased while donor aging.

Key words: periodontal ligament stem cells, aging, proliferation, osteogenic differentiation

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