Evaluation of the synergistic effect of chitosan metal ions (Cu2+/Co2+) in combination with antibiotics to counteract the effects on antibiotic resistant bacteria

Abstract

The effectiveness of antibiotics that save millions of lives is in danger due to the increasing rise of resistant bacteria around the world. We proposed chitosan–copper ions (CSNP–Cu²⁺) and chitosan–cobalt ion nanoparticles (CSNP–Co²⁺) as biodegradable nanoparticles loaded with metal ions synthesized via an ionic gelation method for treatment of antibiotic resistant bacteria. The nanoparticles were characterized using TEM, FT-IR, zeta potential and ICP-OES. The MIC was evaluated for the NPs in addition to evaluating the synergetic effect of the nanoparticles in combination with cefepime or penicillin for five different antibiotic resistant bacterial strains. In order to investigate the mode of action, MRSA, DSMZ 28766 and Escherichia coli E0157:H7 were selected for further evaluation of antibiotic resistant genes expression upon treatment with NPs. Finally, the cytotoxic activities were investigated using MCF7, HEPG2 and A549 and WI-38 cell lines. The results showed quasi spherical shape and mean particle size of 19.9 ± 5 nm, 21 ± 5 nm and 22.27 ± 5 for CSNP, CSNP–Cu²⁺ and CSNP–Co²⁺ respectively. FT-IR showed slight shifting of the hydroxyl and amine group's peaks of chitosan indicating the adsorption of metal ions. Both nanoparticles had antibacterial activity with MIC ranging between 125 and 62 μg ml⁻¹ for the used standard bacterial strains. Moreover, the combination of each of the synthesized NP with either cefepime or penicillin not only showed a synergetic effect as antibacterial activity of each NP or antibiotics alone, but also decreased the fold of antibiotic resistance genes expression. The NPs showed potent cytotoxic activities for MCF-7, HepG2 and A549 cancer cell lines with lower cytotoxic values for the WI-38 normal cell line. The NPs' antibacterial activity may be due to penetration and rupture of the cell membrane and the outer membrane of Gram negative and Gram positive bacteria causing bacterial cell death, in addition to, penetration into the bacterial genes and blocking gene expression that is vital to bacterial growth. The fabricated nanoparticles can be an effective, affordable and biodegradable solution to challenge antibiotic resistant bacteria.