高糖微环境通过重塑细胞外基质影响牙髓损伤修复的研究

doi:10.13701/j.cnki.kqyxyj.2025.10.005

摘要/Abstract

摘要： 目的: 探究高糖微环境中牙髓细胞外基质(extracellular matrix,ECM)的变化及其对牙髓干细胞(dental pulp stem cells,DPSCs)迁移能力与牙髓损伤修复的影响。方法: 选取8周龄雄性Wistar大鼠和GK大鼠,苏木精-伊红染色(hematoxylin-eosin,HE)观察牙髓形态;马松染色和免疫组化染色检测牙髓ECM中胶原纤维含量与交联情况;生物型原子力显微镜检测牙髓ECM的刚度。建立牙髓损伤修复模型,Micro-CT扫描重建和H&E染色评估牙髓损伤修复情况;CD146和STRO-1免疫荧光共染定位DPSCs。免疫荧光染色、划痕实验及Transwell实验检测葡萄糖交联ECM中hDPSCs的形态、迁移能力及力学信号分子的表达。构建ALT-711抑制ECM葡萄糖交联的体内外模型,检测牙髓损伤修复情况与hDPSCs的形态、迁移能力及力学信号分子的表达。结果: 高糖组牙髓截面面积缩小;胶原纤维含量增多、Ⅰ型胶原蛋白(collagenⅠ,COL 1)与晚期糖基化终末产物(advanced glycation end-products,AGEs)表达上调;刚度增加。牙髓损伤后,高糖组未形成修复性牙本质桥、髓腔异位钙化显著,CD146⁺ STRO-1⁺ DPSCs远离损伤部位分布。葡萄糖交联ECM中的hDPSCs面积与形状因子减小、迁移能力下降、ITG-α5β1(integrin alpha 5 beta 1)及Yes相关蛋白(Yes-associated protein,YAP)表达增加。ALT-711干预后,高糖组形成修复性牙本质桥、异位钙化减少,CD146⁺ STRO-1⁺ DPSCs于损伤部位附近分布,hDPSCs形态与迁移能力改善、力学信号减弱。结论: 高糖微环境诱发牙髓ECM胶原异常沉积与过度交联,进而通过激活ITG-YAP力学信号轴导致DPSCs迁移能力障碍及牙髓损伤修复受阻。靶向抑制ECM葡萄糖交联可挽救DPSCs受损的迁移能力和异常的力学信号,并促进牙髓损伤修复。

关键词: 高糖微环境, 细胞外基质, 牙髓损伤修复, 牙髓干细胞, 迁移

Abstract: Objective: To investigate the changes in the extracellular matrix (ECM) of dental pulp in a high glucose microenvironment and its effect on the migratory capacity of dental pulp stem cells (DPSCs) and repair of pulpal injuries. Methods: Eight-week-old male Wistar rats and GK rats were selected and stained with hematoxylin-eosin (HE) to observe the morphology of the dental pulp. Matson staining and immunohistochemical staining were used to detect the collagen fiber content and cross-linking in the pulpal ECM. Bio-type AFM was used to detect the stiffness of the pulpal ECM. A model of pulpal damage repair was established, and Micro-CT scanning reconstruction and HE staining were used to evaluate the repair of pulpal damage. CD146 and STRO-1 immunofluorescence co-staining was used to locate DPSCs. Immunofluorescence staining, scratch assay, and Transwell assay were used to detect the morphology, migration ability, and expression of mechanical signalling molecules of hDPSCs in glucose cross-linked ECM. An in vitro and in vivo model of ALT-711 inhibition of glucose cross-linking in ECM was constructed, and the morphology, migration capacity, and mechanosignalling molecules of hDPSCs were examined in relation to the repair of pulpal injury. Results: In the high glucose group, the pulp cross-sectional area was reduced, collagen fiber content was increased, the expression of collagen I (COL 1) and advanced glycation end-products (AGEs) was up-regulated, and the stiffness was increased. After pulpal injury, the high glucose group did not form restorative dentin bridges, pulpal ectopic calcification was significant, and CD146⁺ STRO-1⁺ DPSCs were distributed far from the injury site. In the glucose cross-linked ECM, hDPSCs had reduced area and shape factor, decreased migration ability, and increased expression of ITG-alpha 5 beta 1 and Yes-associated protein (YAP). hDPSCs in the high-glucose group formed restorative dentin bridges, ectopic calcification was reduced, and CD146⁺ STRO-1⁺ DPSCs were distributed far from the injury site after the ALT-711 intervention. After ALT-711 intervention, the formation of restorative dentin bridges and ectopic calcification were reduced in the high glucose group, CD146⁺ STRO-1⁺ DPSCs were distributed near the injury site, and the morphology and migration of hDPSCs were improved and the mechanical signals were weakened. Conclusion: The high glucose microenvironment induces abnormal deposition and excessive cross-linking of pulpal ECM collagen, which in turn leads to impaired migration of DPSCs and impaired pulpal damage repair through activation of the ITG-YAP mechanical signalling axis. Targeted inhibition of ECM glucose cross-linking can rescue the impaired migration ability and abnormal mechanical signals of DPSCs and promote pulpal injury repair.

Key words: high glucose microenvironment, extracellular matrix, pulp injury and repair, dental pulp stem cells, migration

杜桂林, 刘肖晨, 张旗. 高糖微环境通过重塑细胞外基质影响牙髓损伤修复的研究[J]. 口腔医学研究, 2025, 41(10): 856-866.

DU Guilin, LIU Xiaochen, ZHANG Qi. Study on Role of High Sugar Microenvironment in Affecting Repair of Pulpal Injuries by Remodelling Extracellular Matrix[J]. Journal of Oral Science Research, 2025, 41(10): 856-866.

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