Carboxymethyl chitosan regulates oxidative stress and decreases the expression levels of tumor necrosis factor α in macrophages induced by wear particles.

Abstract

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 4h; (iii) the titanium particles group received stimulation of titanium particles for 12h; and (iv) the CMC group received pre-stimulation of CMC hydrogels for 4h followed by treatment of titanium particles for 12h. 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.