Liu Feng, Pan Hao, Xie Ming, Wang Yingzhen, Xu Hao
Department of Orthopedics, Wuhan Fourth Hospital, Wuhan, 430000 Hubei China.
Department of Joint Surgery, The Affiliated Hospital of Qingdao University, Qingdao, 266000 Shandong China.
Cytotechnology. 2023 Apr;75(2):153-163. doi: 10.1007/s10616-023-00569-z. Epub 2023 Jan 12.
The aim of the present study was to determine the effects of carboxymethyl chitosan (CMC) on titanium particles-induced oxidative stress in mouse RAW264.7 macrophages. The mouse RAW264.7 macrophages were divided into four groups: (i) the control group; (ii) the CMC group received stimulation of CMC for 4 h; (iii) the titanium particles group received stimulation of titanium particles for 12 h; and (iv) the CMC group received pre-stimulation of CMC hydrogels for 4 h followed by treatment of titanium particles for 12 h. Afterwards, reactive oxygen species (ROS) level in the cells was measured by flow cytometry. A spectrophotometer was used to measure the activities of oxidases and antioxidant enzymes. Fluorescence quantitative PCR was performed to analyze mRNA levels of enzymes and tumor necrosis factor α (TNF-α). ELISA was used to detect the mass concentration of TNF-α after indicated treatment. CMC effectively suppressed titanium particles-induced oxidative stress in RAW264.7 cells, as evidenced by the decrease in intracellular ROS level, the transcription of oxidases, and TNF-α concentration as well as the increase in the transcription of antioxidant enzymes. CMC exerts a protective impact against wear particles-induced oxidative stress and reduces the release of TNF-α in RAW264.7 cells.
本研究的目的是确定羧甲基壳聚糖(CMC)对小鼠RAW264.7巨噬细胞中钛颗粒诱导的氧化应激的影响。将小鼠RAW264.7巨噬细胞分为四组:(i)对照组;(ii)CMC组接受CMC刺激4小时;(iii)钛颗粒组接受钛颗粒刺激12小时;(iv)CMC组先接受CMC水凝胶预刺激4小时,然后接受钛颗粒处理12小时。之后,通过流式细胞术测量细胞中的活性氧(ROS)水平。使用分光光度计测量氧化酶和抗氧化酶的活性。进行荧光定量PCR以分析酶和肿瘤坏死因子α(TNF-α)的mRNA水平。使用酶联免疫吸附测定法(ELISA)检测指定处理后TNF-α的质量浓度。CMC有效抑制了RAW264.7细胞中钛颗粒诱导的氧化应激,细胞内ROS水平降低、氧化酶转录和TNF-α浓度降低以及抗氧化酶转录增加证明了这一点。CMC对磨损颗粒诱导的氧化应激具有保护作用,并减少RAW264.7细胞中TNF-α的释放。