Disrupting antimicrobial resistance: unveiling the potential of vitamin C in combating biofilm formation in drug-resistant bacteria.

BACKGROUND

Antimicrobial resistance (AMR) poses a significant threat to global health, exacerbated by the protective mechanisms of biofilms formed by drug-resistant bacteria. Extracellular polymeric substances (EPS) produced by bacteria in biofilms serve as a formidable shield, impeding the efficacy of antimicrobial agents. Here, we investigated the potential of vitamin C (sodium ascorbate) to disrupt biofilm formation in drug-resistant bacteria isolated from diabetic foot ulcer (DFU) patients and studied the antimicrobial and antibiofilm activity of vitamin C on these bacteria.

RESULTS

Out of 117 study isolates, primarily identified as Escherichia coli (n = 52), Staphylococcus spp. (n = 19), and Klebsiella spp. (n = 46), 80 isolates exhibited a Multiple Antimicrobial Resistance (MAR) index greater than 0.2, classifying them as multi-drug resistant (MDR) superbugs. Among these, 58 isolates demonstrated moderate to strong biofilm-forming abilities and were selected for further experiments with vitamin C. The effective concentration of vitamin C inhibiting the growth of most E. coli and Klebsiella spp. isolates (90%) was estimated at 1.25 mg/ml and 2.5 mg/ml respectively, while for allStaphylococcus spp. isolates, it was 0.325 mg/ml. Vitamin C exhibited a notable anti-biofilm effect against the studied isolates, with biofilm prevention concentrations (BPC) of 0.625, 1.25, and 0.16 mg/ml for E. coli, Klebsiella spp., and Staphylococcus spp. isolates respectively. Furthermore, when combined with oxacillin or amoxicillin - drugs that were found ineffective, vitamin C significantly reduced the ability of MDR isolates to form biofilms, rendering them susceptible to the drugs' effects and restoring their efficacy. The expression of the recA gene, an early and quantifiable marker for the onset of the SOS response and biofilm production was downregulated after treatment of E. coli with vitamin C. Relative gene expression analysis revealed that ciprofloxacin-induced recA expression was significantly inhibited when MDR isolates of E. coli were treated with vitamin C at a concentration of 0.625 mg/ml, the BPC of vitamin C.

CONCLUSION

Our findings reveal that vitamin C, alone or in combination with ineffective antibiotics, attenuates biofilm formation and restores the susceptibility of multidrug-resistant (MDR) isolates to antimicrobial agents. This study underscores the promise of vitamin C as a non-lethal disruptor of biofilm-associated antimicrobial resistance.

CLINICAL TRIAL NUMBER: Not applicable.