钛表面纳米形貌对巨噬细胞线粒体代谢及极化方向影响的研究  

The influence of titanium nanotopography on the mitochondrial metabolism and polarization of macrophages

作　　者：赵美琪 于卫华 丁丽 张燕 宋文 张玉梅 ZHAO Meiqi;YU Weihua;DING Li;ZHANG Yan;SONG Wen;ZHANG Yumei(State Key Laboratory of Oral&Maxillofacial Reconstruction and Regeneration,National Clinical Research Center for Oral Diseases,Shaanxi Key Laboratory of Stomatology,The Third Affiliated Hospital of Air Force Medical University,China,710032 Xi’an;School of Military Preventive Medicine,Air Force Medical University,Xian,Shaanxi Key Laboratory of Stomatology,The Third Affiliated Hospital of Air Force Medical University,China,710032 Xi’an;Hospital of Xi’an Shiyou University,Shaanxi Key Laboratory of Stomatology,The Third Affiliated Hospital of Air Force Medical University,China,710032 Xi’an)

机构地区：[1]口颌系统重建与再生全国重点实验室,国家口腔疾病临床医学研究中心,陕西省口腔医学重点实验室,空军军医大学第三附属医院,西安710032 [2]空军军医大学军事预防医学系军事毒理学教研室 [3]西安石油大学医院

出　　处：《实用口腔医学杂志》2025年第1期34-40,共7页Journal of Practical Stomatology

基　　金：国家自然科学基金(编号:32071324)。

摘　　要：目的:探究不同钛表面形貌对巨噬细胞线粒体代谢及极化方向的影响。方法:阳极氧化法在光滑钛(P组)表面制备不同纳米管(NT5组、NT20组)形貌,扫描电镜观察钛试样的表面形貌。在试样表面培养RAW264.7细胞24 h后,扫描电镜观察各组RAW264.7的细胞形态;荧光探针法、荧光素酶法检测各组细胞线粒体代谢产物(ATP、ROS)水平及己糖激酶(HK)活性;RT-qPCR检测各组材料对巨噬细胞极化相关基因(iNOS、Arg-1、TNF-α、CD206)mRNA表达水平。使用2-脱氧-D-葡萄糖(2-DG)抑制P组与NT20组细胞糖酵解后(P^(*)组、NT20^(*)组),再次检测线粒体代谢与巨噬细胞极化的变化。结果:与P组相比,NT5组线粒体ATP、ROS水平及HK活性无明显差异,M2相关基因表达升高;NT20组线粒体ATP、ROS水平上调,HK活性增加,M1相关基因表达升高。抑制糖酵解过程后,与P组相比,P^(*)组与NT20^(*)组线粒体ATP、ROS水平、HK活性及极化相关基因表达水平均无明显差异。结论:大管径纳米管可能通过调控巨噬细胞线粒体发生代谢重编程促进巨噬细胞M1极化。Objective:To explore the effects of different titanium(Ti)surfaces on mitochondria metabolism and polarization of macrophages.Methods:The anodic oxidation method prepared nanotubes with distinct morphologies(NT5 and NT20 groups)on smooth titanium surfaces(P group)were characterized by scanning electron microscopy(SEM).After 24 hours of culturing RAW264.7 cells on the samples,SEM was employed to observe the morphology of the cells of all groups.Levels of mitochondria ATP and ROS,as well as the activity of hexokinase(HK),were measured by fluorescence probes and luciferase assays.The expression of iNOS,Arg-1,TNF-αand CD206 mRNA was evaluated by RT-qPCR.Changes in mitochondria metabolism and macrophage polarization were studied following inhibition of cellular glycolysis by using 2-deoxy-D-glucose(2-DG)in groups P^(*)and NT20^(*).Results:Compared with the P group,the NT5 group showed no significant changes in mitochondria ATP,ROS levels and HK activity,but an upregulation in M2-associated gene expression was observed.In the NT20 group,mitochondria ATP and ROS levels were increased,HK activity was elevated,and M1-associated gene expression was upregulated.After glycolysis inhibition,there was no significant difference in mitochondria ATP,ROS levels,HK activity and polarization-related gene expression levels between the P^(*)and NT20^(*)groups compared with the P group.Conclusion:Large-diameter nanotubes may promote macrophage M1 polarization by regulating the metabolic reprogramming that occurs in macrophage mitochondria.

关 键 词：巨噬细胞极化 二氧化钛纳米管 线粒体 代谢重编程 

分 类 号：R783.1[医药卫生—口腔医学] R783.6[医药卫生—临床医学]