Fabrication of liposome-chitosan-zno nanohybrid integrated with Carissa Spinarum extract for antibacterial application

Abstract

Innovative biomaterials provide a stimulating and adaptable platform for the implementation of new and more effective methods to prevent bacterial infection. Built on biomimetic- inorganic hybrid material, Dual Nanohybrid Delivery System (DN-DS) has advantageous properties for biomedical applications, such as the delivery of herbal formulations for the treatment of bacterial infections. Using microwave assisted extraction (MAE), the polyphenols of Carissa spinarum were extracted. The Dual Nanohybrid Delivery System (LipCsP-ZnONPs)-CT was formed by combining LipCsP-Chitosan and ZnO-Chitosan, which were both generated using different methods of co-precipitation and ion gelation, respectively. A Zetasizer was used to characterize the nanosystems' size, zeta potential, and polydispersity index (PDI). A UV-visible spectrophotometer was utilized for the optical study, and a scanning electron microscope was employed to investigate at the surface morphology. The interaction of coated chitosan with liposomes and ZnONPs was evaluated using Fourier Transformation Infrared (FTIR) spectroscopy. Different kinetic models were fitted to the results of the encapsulation and release profiles of polyphenols in the liposome nanosystems to determine the mechanism of release. Antibacterial activity of the nanoformulations was assessed by an agar diffusion assayand the micro plate blue assay (MABA). The Zeta potential of LipCsP changed from -45.3 ± 0.78 to +34.43 ±1.36 due to chitosan coatings. Polyphenol-encapsulation efficiency was higher in LipCsP-Chitosan (81 ± 2.5%) than in LipCsP (66.11 ± 1.11%). Conversely, the size of LipCsP (176.17 ± 1.05 nm) increasedto 365.2 ± 0.70 nm. FTIR analysis revealed the interaction of the liposome with chitosan due to the disappearance of N-H primary amine. Interaction between chitosan and zinc oxide was revealed by the formation of new absorption peaks at 670 cm-1 and 465 cm-1 as observed in the FTIR analysis. (LipCsP-ZnONPs)-CT presented high bioaccessibility of polyphenols in the simulated gastric phase (82.14 ± 0.80%) than in simulated intestinal phase (71.60 ± 0.86%), a stable system for sustained release of polyphenols, and prominent antibacterial activity. (LipCsP-ZnONPs)-CT exhibited a relative inhibition zone diameter (RIZD) of 89.60 ± 1.32, significant high viability reduction (P˂0.05) against Klebsiella pneumoniae as compared to LipCsP-Chitosan and ZnO-chitosan. The nanohybrid systems (LipCsP-Chitosan and ZnO- chitosan) exhibited synergistic effect against Klebsiella pneumoniae. This study successfully demonstrated the utility of the nanohybrid as a potential antibacterial agent against K.pneumoniae, therefore, the fabricated dual nano delivery system is an efficacy material for treatment of pneumococcal infections.