Antibiofilm Effect of Octenidine Hydrochloride on Staphylococcus aureus, MRSA and VRSA

Abstract

Millions of indwelling devices are implanted in patients every year, and staphylococci (S. aureus, MRSA and vancomycin-resistant S. aureus (VRSA)) are responsible for a majority of infections associated with these devices, thereby leading to treatment failures. Once established, staphylococcal biofilms become resistant to antimicrobial treatment and host response, thereby serving as the etiological agent for recurrent infections. This study investigated the efficacy of octenidine hydrochloride (OH) for inhibiting biofilm synthesis and inactivating fully-formed staphylococcal biofilm on different matrices in the presence and absence of serum protein. Polystyrene plates and stainless steel coupons inoculated with S. aureus, MRSA or VRSA were treated with OH (zero, 0.5, one, 2 mM) at 37 °C for the prevention of biofilm formation. Additionally, the antibiofilm effect of OH (zero, 2.5, five, 10 mM) on fully-formed staphylococcal biofilms on polystyrene plates, stainless steel coupons and urinary catheters was investigated. OH was effective in rapidly inactivating planktonic and biofilm cells of S. aureus, MRSA and VRSA on polystyrene plates, stainless steel coupons and urinary catheters in the presence and absence of serum proteins. The use of two and 10 mM OH completely inactivated S. aureus planktonic cells and biofilm (>6.0 log reduction) on all matrices tested immediately upon exposure. Further, confocal imaging revealed the presence of dead cells and loss in biofilm architecture in the OH-treated samples when compared to intact live biofilm in the control. Results suggest that OH could be applied as an effective antimicrobial to control biofilms of S. aureus, MRSA and VRSA on appropriate hospital surfaces and indwelling devices.

Figure 1

Inhibition of S. aureus (ATCC 35556), vancomycin-resistant S. aureus (VRSA) (VRS 8) and MRSA (NRS 123) biofilm formation on polystyrene by octenidine hydrochloride (OH). Duplicate samples were used for each treatment, and the experiment was replicated three times. Data are represented as the mean ± SEM.

Figure 2

Inhibition of S. aureus (ATCC 35556), VRSA (VRS 8) and MRSA (NRS 123) biofilm on stainless steel by octenidine hydrochloride. Duplicate samples were used for each treatment, and the experiment was replicated three times. Data are represented as the mean ± SEM (Standard Error of Mean).

Figure 3

Inactivation of S. aureus (ATCC 35556), VRSA (VRS 8) and MRSA (NRS 123) biofilm on polystyrene by octenidine hydrochloride. Duplicate samples were used for each treatment, and the experiment was replicated three times. Data are represented as the mean ± SEM(Standard Error of Mean).

Figure 4

Inactivation of S. aureus (ATCC 35556), VRSA (VRS 8) and MRSA (NRS 123) biofilm on stainless steel by octenidine hydrochloride. Duplicate samples were used for each treatment, and the experiment was replicated three times. Data are represented as the mean ± SEM (Standard Error of Mean).

Figure 5

Inactivation of S. aureus (ATCC 35556), VRSA (VRS 8) and MRSA (NRS 123) biofilm on urinary catheters by octenidine hydrochloride. Duplicate samples were used for each treatment, and the experiment was replicated three times. Data are represented as the mean ± SEM (Standard Error of Mean).

Figure 6

Confocal microscopy of MRSA (NRS 385) biofilm without treatment (A) and after treatment with octenidine hydrochloride (B).