中国实用口腔科杂志

中国实用口腔科杂志 ›› 2026, Vol. 19 ›› Issue (4): 451-457.DOI: 10.19538/j.kq.2026.04.011

• 论著 • 上一篇    下一篇

碳化钛纳米片对衰老的人骨髓间充质干细胞影响研究

冯竟朔1，黄超杰2，李明山2，万思杰2，陈    英3，邓旭亮1，3   

  1. 1. 北京大学医学部医学技术研究院，北京 100191；2. 北京航空航天大学化学学院，北京 100191；3. 北京大学口腔医学院·口腔医院特诊科，北京 100081
  • 出版日期:2026-07-30 发布日期:2026-07-30
  • 通讯作者: 邓旭亮，陈英
  • 基金资助:
    国家自然科学基金优秀青年科学基金（52522303）；国家自然科学基金原创探索计划项目延续资助（52550002）；国家自然科学基金面上项目（52373066）；北京市科技新星计划项目（20230484326）；中央高校基本科研业务费（BMU2025PYJH002）

  • Online:2026-07-30 Published:2026-07-30

摘要： 目的    探究碳化钛（titanium carbide，化学式为Ti3C2Tx）纳米片对衰老人骨髓间充质干细胞（human bone marrow mesenchymal stem cells，hBMSCs）的影响，为老年患者重度吸收牙槽骨的再生修复治疗新策略提供实验依据。方法    研究于2025年3月至2026年1月在北京大学医学部实验室进行。采用化学蚀刻法制备Ti3C2Tx纳米片并进行鉴定。通过细胞多次连续传代诱导hBMSCs衰老，对第3代细胞（下文简称“P3”）、第12代细胞（下文简称“P12”）进行β-半乳糖苷酶（senescence-associated β-galactosidase，SA β-Gal）染色和活性氧（reactive oxygen species，ROS）荧光染色分析。采用CCK-8法优化Ti3C2Tx纳米片质量浓度，将其与P12共培养记为Ti3C2Tx组，不加任何刺激的P12记为Ctrl组。对2组衰老相关指标进行检测，包括ROS荧光染色、SA β-Gal染色和磷酸化组蛋白变体γ-H2A.X免疫荧光染色。结果    制备的Ti3C2Tx纳米片为层状结构，横向片径为（7.44 ± 0.18）μm。通过细胞多次连续传代技术，成功构建衰老hBMSCs模型；P12的SA β-Gal阳性细胞率、ROS相对荧光强度均显著高于P3，差异均有统计学意义（均P < 0.05）。与质量浓度0 mg/L比较，Ti3C2Tx纳米片质量浓度为15 mg/L时细胞活力显著增强，差异有统计学意义（t = 15.232，P < 0.001）。Ti3C2Tx组ROS相对荧光强度为10.66 ± 2.97，Ctrl组为70.98 ± 6.51；Ti3C2Tx组SA β-Gal阳性细胞率为（38.22 ± 8.67）%，Ctrl组为（65.65 ± 8.13）%；Ti3C2Tx组γ-H2A.X相对荧光强度为39.74 ± 13.16，Ctrl组为81.78 ± 13.69；以上衰老相关指标组间比较，差异均有统计学意义（均P < 0.05）。结论    质量浓度15 mg/L的Ti3C2Tx纳米片对hBMSCs具有抗衰老作用，其作用机制仍需进一步探究。

关键词: 人骨髓间充质干细胞, 衰老, 活性氧, 碳化钛

Abstract: Objective    To investigate the effects of titanium carbide（Ti3C2Tx）nanosheets on senescent human bone marrow mesenchymal stem cells（hBMSCs），and to provide experimental evidence for developing novel regenerative strategies for the repair of severely resorbed alveolar bone in elderly patients. Methods    This study was conducted in the laboratory of Peking University Health Science Center from March 2025 to January 2026. Ti3C2Tx nanosheets were synthesized using a chemical etching method and subsequently characterized. Cellular senescence in hBMSCs was induced through continuous serial passaging. Passage 3 cells（P3）and passage 12 cells（P12）were evaluated by senescence-associated β-galactosidase（SA β-Gal）staining and reactive oxygen species（ROS）fluorescence staining. The optimal concentration of Ti3C2Tx nanosheets was determined using a CCK-8 assay. P12 cells co-cultured with Ti3C2Tx nanosheets were designated as the Ti3C2Tx group，whereas untreated P12 cells served as the control（Ctrl）group. Senescence-related indicators were assessed in both groups，including ROS fluorescence staining，SA-β-Gal staining，and immunofluorescence staining for the phosphorylated histone variant γ-H2A.X. Results    The prepared Ti3C2Tx nanosheets exhibited a layered structure with a lateral sheet diameter of（7.44 ± 0.18）μm. An aged hBMSC model was successfully established through repeated cell passaging；the SA β-Gal-positive cell rate and relative ROS fluorescence intensity in P12 cells were both significantly higher than those in P3 cells，with all differences being statistically significant（all P < 0.05）. Compared with a concentration of 0 mg/L，cell viability was significantly enhanced at a Ti3C2Tx nanosheet concentration of 15 mg/L，with a statistically significant difference（t = 15.232，P < 0.001）. The relative fluorescence intensity of ROS in the Ti3C2Tx group was 10.66 ± 2.97，while that in the Ctrl group was 70.98 ± 6.51；the SA β-Gal-positive cell rate in the Ti3C2Tx group was（38.22 ± 8.67）%，and that in the Ctrl group was（65.65 ± 8.13）%；the relative fluorescence intensity of γ-H2A.X in the Ti3C2Tx group was 39.74 ± 13.16，while that in the Ctrl group was 81.78 ± 13.69；all of the above intergroup comparisons showed statistically significant differences（all P < 0.05）. Conclusion    Ti3C2Tx nanosheets at a concentration of 15 mg/L exert anti-senescent effects on hBMSCs. However，the underlying mechanisms require further investigation.

Key words: human bone marrow mesenchymal stem cells, aging, reactive oxygen species, titanium carbide