增龄对牙周膜干细胞生物学特性的影响

杜婷婷; 刘娜; 顾斌; 李影; 袁一方; 张维; 张彤

doi:10.24920/J1001-9294.2017.012

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增龄对牙周膜干细胞生物学特性的影响

中国医学科学杂志（英文版） 2017年32卷第2期页码：83-91

Orginal Article | Updated：2024-04-10

- 增龄对牙周膜干细胞生物学特性的影响
- Affiliations：
  
  1. 1 Department of Stomatology, Chinese PLA General Hospital, Beijing 100853, China
  2. 2 Technical Institute of Physics and Chemistry, Beijing 100190, China
- Author bio：
  
  *Corresponding author Tel: 86-10-66937947, E-mail: kqzhengji301@163.com†These authors contributed equally to the work.
- Funds：
  
  △Supported by National Natural Science Foundation of China（51473175）, Science and Technology Nova Plan of Beijing City（Z141107001814101）.
- DOI：10.24920/J1001-9294.2017.012
  中图分类号：
- 收稿日期：2017-01-22，
  
  网络出版日期：2017-06-20，
  
  纸质出版日期：2017-06
- Accepted：
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杜婷婷, 刘娜, 顾斌, 等. 增龄对牙周膜干细胞生物学特性的影响[J]. 中国医学科学杂志（英文版）, 2017,32(2):83-91.

Du Tingting, Liu Na, Gu Bin, et al. Effects of Aging on the Proliferation and Differentiation Capacity of Human Periodontal Ligament Stem Cells^△[J]. Chinese medical sciences journal, 2017, 32(2): 83-91.
杜婷婷, 刘娜, 顾斌, 等. 增龄对牙周膜干细胞生物学特性的影响[J]. 中国医学科学杂志（英文版）, 2017,32(2):83-91. DOI： 10.24920/J1001-9294.2017.012.

Du Tingting, Liu Na, Gu Bin, et al. Effects of Aging on the Proliferation and Differentiation Capacity of Human Periodontal Ligament Stem Cells^△[J]. Chinese medical sciences journal, 2017, 32(2): 83-91. DOI： 10.24920/J1001-9294.2017.012.

摘要

目的

分别获取不同年龄组的人牙周膜干细胞（periodontal ligament stem cells

PDLSC）

对其增殖、分化和凋亡能力进行分析对比

评估增龄因素对其生物学特性的影响。

方法

选取来源于24位不同患者因正畸减数需要拔除的前磨牙24颗

根据患者年龄分为 A组（年龄 18-20岁）、 B组（年龄 30-35岁）、 C组（年龄 45-50岁）。低密度多克隆法分离培养获得PDLSC。分别检测各组细胞的克隆形成能力、碱性磷酸酶( alkaline phosphatase

ALP)活性

流式细胞仪分析细胞周期和凋亡

衰老相关β半乳糖苷酶(Senescence-Associated β-Galactosidase

SA-βG) 染色分析细胞衰老情况;定量RT-PC检测Runx-2、col-1和ALP 等成骨相关基因的表达水平。

结果

不同年龄组供体的牙周膜中均分离出PDLSC

其克隆形成率分别为36.67%

22.67%和9.33% (P

0.05)

随年龄增加而降低;SA-βG 染色随年龄增加阳性染色细胞数目增多

阳性染色细胞数目的百分比分别为4.14%

16.39%和 50.38% (P

0.05);成骨分化能力随年龄增加而降低;G2/S期的比例在A

B和C组分别为 38.73%

29.88%和18.25%(P

0.05)

随年龄增加而降低;凋亡早期与晚期的比例在A

B和C组分别为1.57%

4.56%和5.84% (P

0.05)

随年龄增加而升高;成骨诱导7d后ALP染色和成骨相关基因Runx-2、col-1和ALP的表达也显示出年龄增加相关的下降趋势。

结论

不同年龄组患者的牙周膜中均可分离培养出PDLSC

但其增殖和成骨分化能力随供体年龄的增加而降低。

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 (

0.05). SA-β-G expression of the senescent PDLSC in group A

B and C were 4.14%

16.39%

50.38%

respectively (

0.05). Cells in G2/S phase was 38.73%

29.88%

18.25% (

0.05)

and the apoptosis rate was 1.57%

4.56%

5.84% (

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

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.

关键词

Keywords

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