Stromal Cell-Derived Factor-1 Significantly Induces Proliferation, Migration, and Collagen Type I Expression in a Human Periodontal Ligament Stem Cell Subpopulation
Stromal Cell-Derived Factor-1 Significantly Induces Proliferation, Migration, and Collagen Type I Expression in a Human Periodontal Ligament Stem Cell Subpopulation
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DOI：
10.1902/jop.2011.110201
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发表时间：
2012-03-01
影响因子：
4.3
通讯作者：
Ge, Shaohua
中科院分区：
文献类型：
--
作者：
Du, Lingqian;Yang, Pishan;Ge, Shaohua
背景：趋化因子基质细胞衍生因子-1 （SDF-1）在骨髓间充质干细胞募集和组织再生中的关键作用已被报道。然而，其在人牙周韧带干细胞（PDLSCs）中的作用尚不清楚。PDLSCs被认为是牙周组织再生的候选材料，可用于基于干细胞的牙周组织工程。趋化因子受体在PDLSCs上的表达、趋化因子诱导这些细胞的迁移及其随后在组织修复中的功能可能是牙周组织再生的关键过程。方法：从临床健康的因正畸原因拔出的前磨牙中获得PDL组织，用有限稀释法分离单细胞菌落。免疫细胞化学染色检测间充质干细胞标志物STRO-1的表达。用茜素红染色和油红0染色测定分化电位。实时聚合酶链反应（PCR）和免疫细胞化学染色检测SDF-1受体CXCR4的表达。采用3-（4,5-二甲基噻唑-2-基）-2,5-二苯基溴化四唑试验和溴脱氧尿苷结合试验测定PDLSC亚群的生存能力和增殖能力。实时荧光定量PCR检测I型胶原蛋白和碱性磷酸酶的表达，检测SDF-1对细胞分化的影响。结果：20%的PDL单细胞菌落表达STRO-1阳性，该特异亚群在诱导4周后表达CXCR4阳性，形成矿物质和脂质空泡。当SDF-1浓度在100 ~ 400 ng/mL之间时，可显著促进PDLSC亚群的增殖和迁移。CXCR4中和抗体可阻断细胞增殖和迁移，提示SDF-1通过CXCR4对细胞发挥作用。SDF-1显著促进ⅰ型胶原水平，对碱性磷酸酶水平影响不大。结论：SDF-1可能通过引导PDLSCs进入破坏性牙周组织，促进其活化和增殖，影响这些干细胞的分化等机制，具有促进牙周组织再生的潜力。中华牙周病杂志（英文版）；2009;33(3):379-388。
Background: The pivotal role of chemokine stromal cell derived factor-1 (SDF-1) in bone marrow mesenchymal stem cells recruitment and tissue regeneration has already been reported. However, its roles in human periodontal ligament stem cells (PDLSCs) remain unknown. PDLSCs are regarded as candidates for periodontal tissue regeneration and are used in stem cell based periodontal tissue engineering. The expression of chemokine receptors on PDLSCs and the migration of these cells induced by chemokines and their subsequent function in tissue repair may be a crucial procedure for periodontal tissue regeneration.Methods: PDL tissues were obtained from clinically healthy premolars extracted for orthodontic reasons and used to isolate single-cell colonies by the limited-dilution method. Immunocytochemical staining was used to detect the expression of the mesenchymal stem cell marker STRO-1. Differentiation potentials were assessed by alizarin-red staining and oil-red 0 staining. The expression of SDF-1 receptor CXCR4 was evaluated by real-time polymerase chain reaction (PCR) and immunocytochemical staining. 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and bromodeoxyuridine incorporation assay were used to determine the viability and proliferation of the PDLSC subpopulation. Expression of collagen type I and alkaline phosphatase was detected by real-time PCR to determine the effect of SDF-1 on cells differentiation.Results: Twenty percent of PDL single-cell colonies expressed STRO-1 positively, and this specific subpopulation was positive for CXCR4 and formed minerals and lipid vacuoles after 4 weeks induction. SDF-1 significantly increased proliferation and stimulated the migration of this PDLSC subpopulation at concentrations between 100 and 400 ng/mL. CXCR4 neutralizing antibody could block cell proliferation and migration, suggesting that SDF-1 exerted its effects on cells through CXCR4. SDF-1 promoted collagen type I level significantly but had little effect on alkaline phosphatase level.Conclusion: SDF-1 may have the potential of promoting periodontal tissue regeneration by the mechanism of guiding PDLSCs to destructive periodontal tissue, promoting their activation and proliferation and influencing the differentiation of these stem cells. J Periodontol 2012;83:379-388.