The effect of chitosan on osteoclast viability, bone resorption and radical oxygen production of primary osteoclast culture of mouse bone

Objective: The purpose of this investigation was to determine the effect of chitosan on osteoclast cells by observing cell viability, bone resorption, and radical oxygen species (ROS) production. Methods: Osteoclast cells were obtained from the primary culture of bone marrow mouse. The osteoclast cells were identified by tartrate-resistant acid phosphatase (TRAP) marker both on the cells and the culture medium. The osteoclast cell viability was observed with (3-(4,5-dimethylthiazol- 2-yl)-2,5-diphenyltetrazolium bromide) tetrazolium assay and Bradford assay for total protein medium culture, while ROS production was measured with malondialdehyde (MDA) assay. Slices of cow cortical bone were used as a substrate for osteoclastic resorption and concentrated hydrochloric acid were used to activate resorption and pit formation by any osteoclasts. Results: Osteoclast cells were identified by TRAP marker and chitosan treated group cells showed lower optical density value compared to control (p<0.05) on TRAP assay medium culture. Cell viability indicated lower on chitosan group than control (p<0.05). There was a qualitative difference of the pits formed on the bone surface between the control and the chitosan group. There was a significant difference in MDA (mmol/ml) between the control and the chitosan group with (p<0.05). Conclusion: Based on this research, we conclude that chitosan inhibits the viability of osteoclast cells, decreases ROS production and bone resorption.