Green synthesis of chitosan nanoparticles, optimization, characterization and antibacterial efficacy against multi drug resistant biofilm-forming Acinetobacter baumannii

Chitosan nanoparticles (CNPs) are promising versatile cationic polymeric nanoparticles, which have received growing interest over last few decades. The biocompatibility, biodegradability, environmental safety and non-toxicity of the chitosan nanoparticles makes it preferred for a wide range of biological applications including agriculture, medical and pharmaceutical fields. In this study, CNPs were biosynthesized by aqueous extract of Eucalyptus globulus Labill fresh leaves as bio-reductant. Box-Behnken design in 29 experimental runs was used for optimization of different factors affecting the production of CNPs. The maximum yield of CNPs was 9.91 mg/mL at pH of 4.5, chitosan concentration of 1%, incubation time of 60 min and temperature of 50 °C. The crystallinity, particle size and morphology of the biosynthesized CNPs were characterized. The CNPs possess a positively charged surface of 31.1 mV. The SEM images of the CNPs confirms the formation of spherical form with smooth surface. The TEM images show CNPs were spherical in shape and their size range was between 6.92 and 10.10 nm. X-ray diffraction indicates the high degree of CNPs crystallinity. FTIR analysis revealed various functional groups of organic compounds including NH, NH2, C-H, C-O, C-N, O-H, C-C, C-OH and C-O-C. The thermogravimetric analysis results revealed that CNPs are thermally stable. The antibacterial activity of CNPs was determined against pathogenic multidrug-resistant bacteria, Acinetobacter baumannii. The diameters of the inhibition zones were 12, 16 and 30 mm using the concentrations of 12.5, 25 and 50 mg/mL; respectively. When compared to previous studies, the biosynthesized CNPs produced using an aqueous extract of fresh Eucalyptus globulus Labill leaves have the smallest particle sizes (with a size range between 6.92 and 10.10 nm). Consequently, it is a promising candidate for a diverse range of medical applications and pharmaceutical industries.