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Novel Chitosan-based PH-responsive Lipid-polymer Hybrid Nanovesicles (OLA-LPHVs) For Delivery Of Vancomycin Against Methicillin-resistant Staphylococcus Aureus Infections.

Daniel Zumerkorn Hassan, Calvin A. Omolo, Victoria Oluwaseun Fasiku, Chunderika Mocktar, Thirumala Govender
Published 2020 · Chemistry, Medicine
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The development of novel materials is necessary for adequate delivery of drugs to combat the Methicillin-resistant Staphylococcus aureus (MRSA) burden due to the limitations of conventional methods and challenges associated with antimicrobial resistance. Hence, this study aimed to synthesise a novel oleylamine based zwitterionic lipid (OLA) and explore its potential to formulate chitosan-based pH-responsive lipid-polymer hybrid nanovesicles (VM-OLA-LPHVs1) to deliver VM against MRSA. The OLA was synthesised, and the structure characterised by 1H NMR, 13C NMR, FT-IR and HR-MS. The preliminary biocompatibility of OLA and VM-OLA-LPHVs1 was evaluated on HEK-293, A-549, MCF-7 and HepG-2 cell lines using in vitro cytotoxicity assay. The VM-OLA-LPHVs1 were formulated by ionic gelation method and characterised in order to determine the hydrodynamic diameter (DH), morphology in vitro and in vivo antibacterial efficacy. The result of the in vitro cytotoxicity study revealed cell viability of above 75% in all cell lines when exposed to OLA and VM-OLA-LPHVs1, thus indicating their biosafety. The VM-OLA-LPHVs1 had a DH, polydispersity index (PDI), and EE% of 198.0 ± 14.04 nm, 0.137 ± 0.02, and 45.61 ± 0.54% respectively at physiological pH, with surface-charge (ζ) switching from negative at pH 7.4 to positive at pH 6.0. The VM release from the VM-OLA-LPHVs1 was faster at pH 6.0 compared to physiological pH, with 97% release after 72-h. The VM-OLA-LPHVs1 had a lower minimum inhibitory concentration (MIC) value of 0.59 μg/mL at pH 6.0 compared to 2.39 μg/mL at pH 7.4, against MRSA with 52.9-fold antibacterial enhancement. The flow cytometry study revealed that VM-OLA-LPHVs1 had similar bactericidal efficacy on MRSA compared to bare VM, despite an 8-fold lower VM concentration in the nanovesicles. Additionally, fluorescence microscopy study showed the ability of the VM-OLA-LPHVs1 to eliminate biofilms. The electrical conductivity, and protein/DNA concentration, increased and decreased respectively, as compared to bare VM which indicated greater MRSA membrane damage. The in vivo studies in a BALB/c mouse-infected skin model treated with VM-OLA-LPHVs1 revealed 95-fold lower MRSA burden compared to the group treated with bare VM. These findings suggest that OLA can be used as an effective novel material for complexation with biodegradable polymer chitosan (CHs) to form pH-responsive VM-OLA-LPHVs1 nanovesicles which show greater potential for enhancement and improvement of treatment of bacterial infections.
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