Interferon Gamma-treated Dental Pulp Stem Cells Promote Human Mesenchymal Stem Cell Migration In Vitro.
Interferon Gamma-treated Dental Pulp Stem Cells Promote Human Mesenchymal Stem Cell Migration In Vitro.
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DOI：
10.1016/j.joen.2015.02.018
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发表时间：
2015-08
影响因子：
4.2
通讯作者：
Alapati S
中科院分区：
文献类型：
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作者：
Strojny C;Boyle M;Bartholomew A;Sundivakkam P;Alapati S
慢性炎症通过破坏修复的祖细胞募集过程来破坏牙髓再生。骨髓间充质干细胞（BMMSC）与牙髓干细胞（DPSC）具有共同的特征。该研究的目的是研究BM-MSC在炎症趋化剂的作用下向DPSC的迁移。此外，我们的研究还描述了骨髓间充质干细胞在体外介导DPSC增殖和分化的信号机制。使用传代2和传代4之间的人DPSC和BM-MSC，并在成牙分化培养基中生长。用茜素红染色法测定矿化度。用结晶紫染色对生长在Boyden室transwell插入物中的细胞进行迁移评估。用碱性磷酸酶（ALP）活性测定法评价DPSC的矿化潜力。采用Real - time PCR分析cxcl3、5、6、10、11、12、14、16、SDF-α、血管内皮生长因子（VEGF）和成纤维细胞生长因子（FGF）的基因表达谱。茜素红和碱性磷酸酶活性分析显示，IFN-γ处理显著降低了DPSC的分化潜力。当与IFN-γ处理的DPSC共培养时，BM-MSC的迁移增加。RNA表达研究表明，与IFN-γ处理的DPSC共培养时，BMMSC中Cxcl6和Cxcl12的水平升高。此外，在暴露于IFN-γ的DPSC中，观察到促血管生成因子VEGF和FGF的上调。我们的研究结果表明，炎症IFN-γ处理的DPSC释放因子（可能是Cxcl6和12）有助于MSC的归巢。该模型可能为研究MSC-DPSC串音提供了潜在的研究工具，并为未来研究祖干细胞的募集和可持续性维持炎症级联治疗牙髓炎症提供了潜在的研究工具。
Chronic inflammation disrupts dental pulp regeneration by disintegrating the progenitors recruitment process for repair. Bone marrow-derived mesenchymal stem cells (BMMSC) share the common features with dental pulp stem cells (DPSC). The aim of the study was to investigate the migration of BM-MSC towards DPSC, in response to inflammatory chemoattractants. Additionally, our studies also delineated the signaling mechanisms from BMMSC in mediating the proliferation and differentiation of DPSC, in vitro. Human DPSC and BM-MSC between passages 2 and 4 were used and were grown in odontogenic differentiation medium. Mineralization was determined by Alizarin Red staining analysis. Migration was assessed using crystal violet staining in cells grown in Boyden chamber transwell inserts. Mineralization potential of DPSC was evaluated using alkaline phosphatase (ALP) activity assay. Real time PCR analysis was performed to assess the gene expression profile of Cxcl 3, 5, 6, 10, 11, 12, 14, 16, SDF-α, vascular endothelial growth factor (VEGF) and fibroblast growth factor (FGF). IFN-γ treatment significantly abrogated the differentiation potential of DPSC, as shown using alizarin red and alkaline phosphatase activity analysis. An increase in the migration of BM-MSC was documented when co-cultured with IFN-γ-treated DPSC. RNA expression studies showed an increased in the levels of Cxcl6 and Cxcl12 in BMMSC when co-cultured with IFN-γ-treated DPSC. Additionally, an upregulation of proangiogenic factors, VEGF and FGF were observed in DPSC exposed to IFN-γ. Our findings indicate that inflamed IFN-γ-treated DPSC release factors (presumably Cxcl6 and 12) that contribute to the homing of MSC. This model might provide a potential research tool for studying MSC-DPSC cross-talk, and for future studies involving recruitment and sustainability of progenitor stem cells sustaining inflammatory cascade to treat pulp inflammation.