Department of Orthodontics, Dental Clinic, University of Bonn, Bonn, Germany.
J Periodontal Res. 2010 Dec;45(6):764-71. doi: 10.1111/j.1600-0765.2010.01297.x.
Aging modulates the proliferative activity and organic matrix production of cells in vivo and in vitro. Here, we explore how aging affects the phenotypic characteristics of human periodontal ligament cells and their response to hormonal stimulation.
Fifth passage periodontal ligament cells from subjects aged 12-14 (group 1), 41-55 (group 2) and 61-70 years (group 3) were characterized for the expression of mesenchymal marker genes and proteins by real-time PCR and flow cytometry. Confluent cultures were exposed to 10(-12) m parathyroid hormone(1-34) [PTH(1-34)] intermittently for three cycles. At harvest, cell number, alkaline phosphatase activity and osteocalcin production were determined by cell count, biochemical assay and ELISA.
The characterization of the cells revealed a decreased expression of osteoblast-specific marker genes along with a lower percentage of cells presenting the respective proteins with age. An intermittent exposure of the cultures to 10(-12) m PTH(1-34) induced an increase of the cell number as opposed to a significant decrease of alkaline phosphatase activity and osteocalcin production. The cellular response to PTH(1-34) was strongest in group 1. Basal osteoprotegerin levels were highest in the cultures from the oldest donors and inhibited by intermittent PTH(1-34) in all groups.
Our data indicate that periodontal ligament cells from older subjects display a less differentiated phenotype and a reduced response to intermittent PTH, suggesting a compromised ability to maintain tissue homeostasis and a limited possibility to support periodontal repair processes with age. The high basal osteoprotegerin expression in older subjects might serve as a compensatory mechanism.
衰老可调节体内和体外细胞的增殖活性和有机基质产生。在此,我们探讨衰老如何影响人牙周韧带细胞的表型特征及其对激素刺激的反应。
通过实时 PCR 和流式细胞术,对来自 12-14 岁(第 1 组)、41-55 岁(第 2 组)和 61-70 岁(第 3 组)受试者的第 5 代牙周韧带细胞进行间充质标志物基因和蛋白的表达特征分析。当细胞汇合后,将它们间歇性暴露于 10(-12) m 甲状旁腺激素(1-34) [PTH(1-34)]中,共三个周期。收获时,通过细胞计数、生化测定和 ELISA 测定细胞数量、碱性磷酸酶活性和骨钙素产生。
细胞特征分析显示,随着年龄的增长,成骨细胞特异性标志物基因的表达降低,呈现相应蛋白的细胞比例也降低。与碱性磷酸酶活性和骨钙素产生的显著下降相反,间歇性暴露于 10(-12) m PTH(1-34)会增加细胞数量。第 1 组对 PTH(1-34)的细胞反应最强。在最年长供体的培养物中,基础骨保护素水平最高,并在所有组中被间歇性 PTH(1-34)抑制。
我们的数据表明,来自老年受试者的牙周韧带细胞表现出分化程度较低的表型,对间歇性 PTH 的反应减弱,这表明随着年龄的增长,维持组织内稳态的能力受损,支持牙周修复过程的可能性有限。老年受试者中基础骨保护素的高表达可能是一种代偿机制。