Assessing an effect of a new chlorhexidine-containing drug on the functional activity of EA.hy926 line endothelial cells

Chlorhexidine is a drug widely used for wound care, treatment of burns (0.05% solution), as an ingredient in mouth rinses to prevent biofilm formation on tooth surfaces (0.12% solution), as well as in gynecology and otolaryngology. Chlorhexidine is a cationic antiseptic that, due to its positive charge, can cause damage to pathogens’ negatively charged cell wall. Chlorhexidine is effective against both Gram-positive and Gram-negative bacteria; however, it is ineffective against spore-forming bacteria and certain viruses. A significant issue is the reduced microbial sensitivity to chlorhexidine and cross-resistance development. To enhance its antiseptic properties, chlorhexidine can be used in combination with other agents, such as ethanol. Antiseptic agents should demonstrate sufficient efficacy against fungal, bacterial, and viral pathogens; moreover, antiseptics capable of accelerating tissue repair would offer a distinct advantage. Of particular interest is a formulation containing a chlorhexidine-EDTA-zinc complex. EDTA is used to potentiate the action of many antiseptic agents by increasing bacterial cell wall permeability through disruption of lipopolysaccharides, whereas zinc ions exhibit antibacterial and antifungal activity. Recent research has demonstrated that the chlorhexidine-EDTA-zinc complex exhibits high stability and more prominent biocidal properties compared to chlorhexidine bigluconate. Literature data suggest that EDTA can reverse contact inhibition of endothelial cells and stimulate their proliferation. Thus, the chlorhexidine-EDTA-zinc complex potentially combines bactericidal properties along with the ability to accelerate wound healing by stimulating endothelial cells. In the present study, the effect of a chlorhexidine-EDTA-zinc complex-containing formulation on endothelial cell functional activity was evaluated. The cytotoxicity of the formulation was assessed, along with dilution-specific effects on endothelial cell proliferation and migration. This study demonstrated that at certain dilutions chlorhexidine-containing formulation do not affect endothelial cell proliferation but promote an increase in the surface area occupied by the cells. Under such “spreading,” endothelial cells form more intercellular contacts and stronger adhesion to the substrate, leading to stabilization of blood vessels during angiogenesis and improved tissue perfusion and wound healing. Thus, the chlorhexidine-EDTA-zinc complex can be considered as a modern alternative to chlorhexidine bigluconate for wound care.