研究目的
To overcome multidrug resistance in bacteria via inhibition of efflux pump and photoinhibition using thiolated chitosan coated cobalt doped zinc oxide nanoparticles (Co-ZnO) in visible light, specifically against methicillin resistant Staphylococcus aureus (MRSA).
研究成果
Thiolated chitosan coated Co-ZnO nanoparticles effectively kill MRSA through efflux pump inhibition and oxidative stress upon light activation, with high biocompatibility. They show promise as an alternative to conventional antibiotics for treating multidrug-resistant bacterial infections, and future studies should explore their application against other resistant bacteria and in vivo models.
研究不足
The study is limited to in-vitro experiments on MRSA and human red blood cells; in-vivo efficacy and long-term toxicity are not assessed. The mechanism of photothermal effect and detailed ROS interactions could be further optimized. The nanoparticles' performance against other MDR bacteria was not evaluated in this work.
1：Experimental Design and Method Selection:
The study used a modified co-precipitation method to synthesize Co-ZnO nanoparticles, with characterization via XRD and TEM. Antibacterial efficacy was assessed through zone of inhibition and MIC assays, efflux pump inhibition via cart wheel assay, hemolysis for biocompatibility, photothermal activity measurement, and ROS quantification using DPBF assay.
2：Sample Selection and Data Sources:
MRSA strains previously isolated from Holy Family Hospital, Rawalpindi, Pakistan, were used. Human red blood cells from a volunteer were used for hemolysis tests.
3：List of Experimental Equipment and Materials:
Instruments included sonicator (Elmasonic, China), incubator (Panasonic, Japan), spectrophotometer (Thermo Scientific Multiscan Go, USA), and others. Chemicals included zinc nitrate hexahydrate, cobalt nitrate hexahydrate, KOH, etc.
4：Experimental Procedures and Operational Workflow:
Synthesis involved mixing zinc and cobalt nitrates with KOH, centrifugation, drying, and calcination. Coating with thiolated chitosan was done overnight. Antimicrobial tests involved exposing MRSA to nanoparticles in sunlight or dark, measuring zones of inhibition and MIC. Efflux inhibition used ethidium bromide and cart wheel pattern. Hemolysis involved incubating RBCs with nanoparticles and measuring absorbance. Photothermal activity was measured by temperature rise in sunlight. ROS quantification used DPBF dye and UV spectrophotometry.
5：Data Analysis Methods:
Data were analyzed using Origin Pro-8 software, with statistical differences assessed via student t-test (P < 0.05 significant).
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