article · Scientific Reports
Chitosan nanoparticles offer useful antimicrobial and drug delivery features. An alternative, biological synthesis method was developed using olive leaf extract, optimised via experimental design and artificial neural networks. The highest yield reached 21.15 milligrams per millilitre under specific conditions of temperature, acidity, incubation time, and precursor concentrations. Characterisation showed thermally stable, positively charged, crystalline nanoparticles with spherical shapes and sizes ranging between 6.91 and 11.14 nanometres. Laboratory testing revealed that these biosynthesised nanoparticles suppressed biofilm formation by common pathogens, specifically Pseudomonas aeruginosa, Staphylococcus aureus, and Candida albicans, in a dose-dependent manner. This biofilm inhibition corresponded with reductions in cellular metabolic activity, cell hydrophobicity, and protective structural components. The resulting nanoparticles show potential for medical and pharmaceutical uses against drug-resistant microbes.
Biofilms formed by bacteria and fungi shield pathogens from conventional antimicrobials, complicating healthcare management for vulnerable individuals. Using plant extracts to produce ultra-small chitosan nanoparticles offers an eco-friendly manufacturing route. Because these nanoparticles disrupt both the protective matrix and the metabolic activity of problematic microbes in laboratory settings, they provide a foundation for designing alternative therapies to tackle persistent, drug-resistant infections.
This work demonstrates an early-stage, laboratory-tested synthesis process relevant to pharmaceutical and medical technology developers. The primary application is targeting multidrug-resistant pathogens and microbial biofilms, including those associated with secondary pneumonia. Because the evaluation is limited to in vitro assays and predictive yield optimisation, substantial translational research, including formulation scale-up, safety profiling, and in vivo efficacy testing, remains necessary before the technology can approach commercial adoption.
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Abstract Chitosan nanoparticles (CNPs) are promising biopolymeric nanoparticles with excellent physicochemical, antimicrobial, and biological properties. CNPs have a wide range of applications due to their unique characteristics, including plant growth promotion and protection, drug delivery, antimicrobials, and encapsulation. The current study describes an alternative, biologically-based strategy for CNPs biosynthesis using Olea europaea leaves extract. Face centered central composite design (FCCCD), with 50 experiments was used for optimization of CNPs biosynthesis. The artificial neural network (ANN) was employed for analyzing, validating, and predicting CNPs biosynthesis using Olea europaea leaves extract. Using the desirability function, the optimum conditions for maximum CNPs biosynthesis were determined theoretically and verified experimentally. The highest experimental yield of CNPs (21.15 mg CNPs/mL) was obtained using chitosan solution of 1%, leaves extract solution of 100%, initial pH 4.47, and incubation time of 60 min at 53.83°C. The SEM and TEM images revealed that CNPs had a spherical form and varied in size between 6.91 and 11.14 nm. X-ray diffraction demonstrates the crystalline nature of CNPs. The surface of the CNPs is positively charged, having a Zeta potential of 33.1 mV. FTIR analysis revealed various functional groups including C–H, C–O, CONH 2 , NH 2 , C–OH and C–O–C. The thermogravimetric investigation indicated that CNPs are thermally stable. The CNPs were able to suppress biofilm formation by P. aeruginosa, S. aureus and C. albicans at concentrations ranging from 10 to 1500 µg/mL in a dose-dependent manner. Inhibition of biofilm formation was associated with suppression of metabolic activity, protein/exopolysaccharide moieties, and hydrophobicity of biofilm encased cells (r ˃ 0.9, P = 0.00). Due to their small size, in the range of 6.91 to 11.14 nm, CNPs produced using Olea europaea leaves extract are promising for applications in the medical and pharmaceutical industries, in addition to their potential application in controlling multidrug-resistant microorganisms, especially those associated with post COVID-19 pneumonia in immunosuppressed patients.
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DOI: 10.1038/s41598-023-30911-6
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