Green synthesis of polyethylene glycol coated, ciprofloxacin loaded CuO nanoparticles and its antibacterial activity against Staphylococcus aureus

Abstract

Antibacterial resistance requires an advanced strategy to increase the efficacy of current therapeutics in addition to the synthesis of new generations of antibiotics. In this study, copper oxide nanoparticles (CuO-NPs) were green synthesized using Moringa oleifera root extract. CuO-NPs fabricated into a form of aspartic acid-ciprofloxacin-polyethylene glycol coated copper oxide-nanotherapeutics (CIP-PEG-CuO) to improve the antibacterial activity of NPs and the efficacy of the drug with controlled cytotoxicity. These NPs were charachterized by Fourier transform infrared spectroscopy (FTIR), x-rays diffraction spectroscopy (XRD), scanning electron microscopy (SEM) and energy-dispersive spectroscopy (EDS). Antibacterial screening and bacterial chemotaxis investigations demonstrated that CIP-PEG-CuO NPs show enhanced antibacterial potential against Gram-positive and Gram-negative clinically isolated pathogenic bacterial strains as compared to CuO-NPs. In ex-vivo cytotoxicity CIP-PEG-CuO-nano-formulates revealed 88% viability of Baby Hamster Kidney 21 cell lines and 90% RBCs remained intact with nano-formulations during hemolysis assay. An in-vivo studies on animal models show that Staphylococcus aureus were eradicated by this newly developed formulate from the infected skin and showed wound-healing properties. By using specially designed nanoparticles that are engineered to precisely transport antimicrobial agents, these efficient nano-drug delivery systems can target localized infections, ensure targeted delivery, enhance efficacy through increased drug penetration through physical barriers, and reduce systemic side effects for more effective treatment.

Keywords: Staphylococcus aureus; Antibacterial; CIP-PEG-CuO-nanotherapeutic; Green synthesis; Nano-drug delivery.

Fig. 1

XRD analysis of green synthesized CuO-NPs (red) and CIP-PEG-CuO-nano formulate (black).

Fig. 2

(a) SEM images of green synthesized CuO-NPs, (b) EDS of green-CuO-NPs for chemical composition showing the highest peak and percentage of copper.

Fig. 3

Zone of inhibition using CuO-NPs/Nano-formulations against bacterial strains. The mean values with standard deviation were noticed following the triplicate experiment. The significant difference (p < 0.05) calculated by one-way ANOVA between all tested nanomaterials.

Fig. 4

Sensitivity patterns of green synthesized CuO- NPs/nano-formulations against bacterial strains, (a) E. coli, (b) Enterococcus faecalis, (c) Klebsiella pneumonia, (d) MRSA, (e) Pseudomonas aeruginosa, (f) Salmonella typhi, (g) S. aureus.

Fig. 5

(A) Biofilm inhibition and (B) biofilm destruction of Gr-CIP-PEG.CuO-NPs against bacterial strains. The mean values with standard deviation were evaluated. The significant difference (p < 0.05) measured by one-way ANOVA between all tested nanomaterials.

Fig. 6

Antioxidant assay of CuO-NPs and their nano-formulations. The mean values with standard deviation were evaluated. The significant difference (p < 0.05) observed by one-way ANOVA between all tested nanomaterials.

Fig. 7

(A) DNA release study from bacteria with the treatment of CuO-NPs and their nanoformulations, (B) protein leakage study from bacteria with the treatment of CuO-NPs and their nanoformulations. The mean values with standard deviation were evaluated. The significant difference (p < 0.05) measured by one-way ANOVA between all tested nanomaterials.

Fig. 8

(A) The optical density of bacterial chemotaxis toward Gr-CIP-PEG-CuO-NPs/L-aspartic acid, (B) optical density without any bacterial attractant.

Fig. 9

Ex-vivo cytotoxicity of green synthesized CuO-NPs and Gr-CIP-PEG-CuO-NPs on BHK21; (A) (negative control), (B) (positive control); (C) (G-CIP-PEG-Cuo-NPs), (D) ( G-CuO-Nps) E (PEG), F (CIP). The mean values with standard deviation were evaluated. The significant difference (p < 0.05) measured by one-way ANOVA between all tested nanomaterials.

Fig. 10

(a) Wound healing progresses using Gr-CuO-NPs-based nano-therapeutics (b) Relationship between wound size and time using Gr-CuO-NPs-based nano-therapeutics ofr wound healing.

Fig. 11

(a) Wound healing infected by Staphylococcus aureus using Gr-CuO-NPs-based nano-therapeutics (b) Relationship between wound size or infection removal and time using Gr-CuO-NPs-based nano-therapeutics ofr wound healing in treatment-II.

Fig. 12

H & E microscopy of skin tissues (40X), (A) Wound healing after treatment with CIP-PEG-CuO-NPs at 40X, (B) Control model of wound healing without treatment, (C) Infection recovery experimental skin tissue after treatment with CIP-PEG-CuO-NPs, (D) Control model of infection recovery without treatment.