Effects of Zinc Oxide Nanoparticles Synthesized Using Aspergillus niger on Carbapenem-Resistant Klebsiella pneumonia In Vitro and In Vivo

Abstract

Currently, the mortality rate in Saudi Arabia’s ICUs is increasing due to the spread of Klebsiella pneumoniae carbapenemase (KPC)-producing bacteria. This study was carried out to evaluate the ability of biologically synthesized zinc oxide nanoparticles (ZnO-NPs) using Aspergillus niger to overcome carbapenem-resistant K. pneumoniae (KPC) in vitro and in vivo. ZnO-NPs were synthesized via a biological method and characterized using UV–Vis spectroscopy, Zetasizer and zeta potential analyses, x-ray diffraction spectroscopy, Fourier transform infrared spectroscopy, scanning electron microscopy (SEM), and energy-dispersive x-ray spectroscopy (EDX). In vitro sensitivity of KPC to ZnO-NPs was identified using the well diffusion method. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined by a macro-dilution method. The morphological alteration of KPC cells after ZnO-NPs treatment was observed by SEM. The in vivo susceptibility of KPC cells to ZnO-NPs ointment was evaluated using wound healing in experimental rats. The chemical characterization findings showed the formation, stability, shape, and size of the synthesized nanoparticles. The MIC and MBC were 0.7 and 1.8 mg/ml, respectively. The in vivo results displayed reduced inflammation and wound re-epithelialization of KPC-infected rats. These findings demonstrated that ZnO-NPs have great potential to be developed as antibacterial agents.

Molecular Dynamic Analysis of Carbapenem-Resistant Klebsiella pneumonia’s Porin Proteins with Beta Lactam Antibiotics and Zinc Oxide Nanoparticles

“…ZnO-NPs’ MIC presented the lowest MIC and MBC when compared with the tested antibiotics: 0.2 mg/mL, 0.5 mg/mL MIC and MBC, respectively, against KP and KPC; >64 mg/mL MIC and MBC for cefepime against KPC; >16 mg/mL for meropenem’s MIC and MBC against both tested bacteria; and >4 mg/mL MIC and MBC for imipenem against KPC and KP ( Table 4 ). The MIC and MBC results for ZnO-NPs observed in this investigation are far below those observed by our previous study [ 6 , 11 ]. The final in vitro test was the study of bacterial surface alteration before and after treatment by ZnO-NPs and antibiotics using SEM ( Figure 10 A–C).…”

“…ZnO-NPs’ absorbance spectrum revealed a significant absorbance peak at 367.5 nm ( Figure 1 A). This finding is in agreement with previous research findings that zinc oxide nanoparticle formation occurs at sizes between 300 and 390 nm [ 11 , 16 , 17 ]. The mean Z-average diameter (nm) was used as a preliminary screening to evaluate the size of ZnO-NPs.…”

“…Most of these particles had a smooth surface and appeared to be free of fractures. These results reflect those of Elsayim and others, who also found that their synthesized ZnO-NPs had aggregation features and a quaternary shape [ 11 ]. Although Elsayim and others synthesized their nanoparticles by Acacia nilotica , their nanoparticles presented a hexagonal shape, an outcome that was contrary to our result [ 6 ].…”

“…Additionally, neither meropenem nor imipenem exhibited any activity against KP and KPC. This finding was also reported by many researchers who tested ZnO-NPs against carbapenem-resistant Klebsiella pneumoniae [ 6 , 11 , 33 ]. The MIC of an antibacterial agent is measured in mg/L (g/mL), the lowest concentration at which the test strain of an organism cannot grow in any way that can be observed [ 34 ].…”

“…Recently, there has been extensive research conducted on the biological synthesis of nanoparticles to overcome resistant microbes. Zinc oxide nanoparticles are one of the most potent nanoparticles, reported by Elsayim and others as promising antimicrobial compounds [ 11 , 12 , 13 ]. In this research, we intended to evaluate the activity of biosynthesized ZnONPs against KPC in vitro and in silico.…”