Effects of Chlorhexidine-Iodophor Composite Solution on the Viability and Proliferation of Human Skin Fibroblasts Infected by S. aureus – An in Vitro Experiment

Abstract

Background: The chlorhexidine-iodophor (CHX-IP) composite solution is a polymer of chlorhexidine and iodophor, applicable to the control of local microbial load and probably toxic to fibroblasts. However, the effect of CHX-IP on the viability and proliferation of human skin fibroblasts infected by Staphylococcus aureus (S. aureus) remains unknown. Objective: The effects of CHX-IP composite solution on the viability and proliferation of human skin fibroblasts infected by S. aureus were investigated in vitro cell culture in this study. Methods: Optimum multiplicity of infection (MOI) was determined to construct the S. aureus-fibroblast co-culture model. Cell Viability Assay was applied to obtain optical density (OD) value and calculate cell viability. 5-ethynyl-2′- deoxyuridine (EdU) assay was used to investigate the effect of CHX-IP on the proliferation of human skin fibroblasts infected by S. aureus. Results: 10:1 was the optimum MOI for the S. aureus-fibroblast co-culture model. The OD value of human skin fibroblasts infected by S. aureus increased in the blank control group, 0.625 mg/ml, 0.3125 mg/ml, 0.15625 mg/ml, and 0.075625 mg/ml groups after four hours. While that of the negative control group, 5 mg/ml, 2.5 mg/ml, and 1.25 mg/ml groups decreased over time. The two-way ANOVA results indicated that the OD value of human skin fibroblasts infected by S. aureus was significantly different among different CHX-IP concentration groups (F = 34.05, P < .001), and the interaction effect between concentration and time was significant (F = 9.442, P < .001). The results of the EdU cell proliferation assay showed that the blank control group, 0.625 mg/ml CHX-IP group, and 0.075625 mg/ml CHX-IP group had an enhanced fibroblasts cell proliferation, while the fibroblasts cell proliferation of the negative control group and 5 mg/ml CHX-IP group was inhibited. Conclusion: The viability and proliferation of human skin fibroblasts infected by S. aureus were inhibited, while specific concentrations of CHX-IP solution can counteract or even reverse the proliferation inhibition effect.