Silver nanoparticles (AgNPs) are excellent antimicrobial agents and promising candidates for preventing or treating bacterial infections caused by antibiotic resistant strains. However, their increasing use in commercial products raises concerns about their environmental impact. In addition, traditional physicochemical approaches often involve harmful agents and excessive energy consumption, resulting in AgNPs with short-term colloidal stability and silver ion leaching. To address these issues, we designed stable hybrid lignin-silver nanoparticles (AgLigNPs) intended to effectively hit bacterial envelopes as a main antimicrobial target. The lignin nanoparticles (LigNPs), serving as a reducing and stabilizing agent for AgNPs, have a median size of 256 nm and a circularity of 0.985. These LigNPs were prepared using the dialysis solvent exchange method, producing spherical particles stable under alkaline conditions and featuring reducing groups oriented toward a wrinkled surface, facilitating AgNPs synthesis and attachment. Maximum accumulation of silver on the LigNP surface was observed at a mass reaction ratio mAg:mLig of 0.25, at pH 11. The AgLigNPs completely inhibited suspension growth and reduced biofilm development by 50% in three tested strains of Pseudomonas aeruginosa at a concentration of 80/9.5 (lignin/silver) mg L-1. Compared to unattached AgNPs, AgLigNPs required two to eight times lower silver concentrations to achieve complete inhibition. Additionally, our silver-containing nanosystems were effective against bacteria at safe concentrations in HEK-293 and HaCaT tissue cultures. Stability experiments revealed that the nanosystems tend to aggregate in media used for bacterial cell cultures but remain stable in media used for tissue cultures. In all tested media, the nanoparticles retained their integrity, and the presence of lignin facilitated the prevention of silver ions from leaching. Overall, our data demonstrate the suitability of AgLigNPs for further valorization in the biomedical sector.

• Publication type
• Journal Article MeSH

The escalating antibiotic resistance observed in bacteria poses a significant threat to society, with the global prevalence of resistant strains of Pseudomonas aeruginosa on the rise. Addressing this challenge necessitates exploring strategies that would complement existing antimicrobial agents, e.g. by substances mitigating bacterial virulence without eliciting selective pressure for resistance emergence. In this respect, free-form chitosan has demonstrated promising efficacy, prompting our investigation into reinforcing its effects through nanoparticle formulations. Our study focuses on the preparation of chitosan nanoparticles under suitable conditions while emphasizing the challenges associated with stability that can affect biological activity. These challenges are mitigated by introducing quaternized chitosan, which ensures colloidal stability in the culture media. Our approach led to the production of trimethylchitosan nanoparticles with a median size of 103 nm, circularity of 0.967, and a charge of 14.9 ± 3.1 mV, stable within a one-month period in a water stock solution, showing promising attributes for further valorization. Furthermore, the study delves into the antimicrobial activity of trimethylchitosan nanoparticles on Pseudomonas aeruginosa and confirms the benefits of both nanoformulation and modification of chitosan, as our prepared nanoparticles inhibit 50% of the bacterial population at concentration ≥160 mg L-1 within tested strains. Additionally, we identified a concentration of 5 mg L-1 that no longer impedes bacterial growth, allowing reliable verification of the effect of the prepared nanoparticles on Pseudomonas aeruginosa virulence factors, including motility, protease activity, hemolytic activity, rhamnolipids, pyocyanin, and biofilm production. Although trimethylchitosan nanoparticles exhibit promise as an effective antibiofilm agent (reducing biofilm development by 50% at concentrations ranging from 80 to 160 mg L-1) their impact on virulence manifestation is likely not directly associated with quorum sensing. Instead, it can probably be attributed to non-specific interactions with the bacterial surface. This exploration provides valuable insights into the potential of quaternized chitosan nanoparticles in addressing Pseudomonas aeruginosa infections and underscores the multifaceted nature of their antimicrobial effects.

• Publication type
• Journal Article MeSH

The utilization of waste materials for the synthesis of nanoparticles has gained significant attention due to its potential for waste valorization and contribution to circular economy. In this study, bimetallic nanoparticles were produced using extracts derived from Cannabis sativa and Vitis vinifera waste, focusing on their green synthesis and antimicrobial activity against Gram-negative bacteria, specifically several strains of Pseudomonas aeruginosa. The Vitis vinifera canes and post-extraction waste from Cannabis sativa were processed using an ethanol extraction method. The extract was then mixed with silver nitrate and tetrachloroauric acid solution at different reagent ratios to optimize the synthesis process. The resulting bimetallic nanoparticles (AgAuNPs) were characterized using UV-vis spectrophotometry, transmission electron microscopy, atomic absorption spectrometry, X-ray diffraction, X-ray photoelectron spectroscopy and Fourier transform infrared spectroscopy. The antimicrobial activity of the biosynthesized AgAuNPs was evaluated against various strains of Pseudomonas aeruginosa. The minimal inhibitory concentration (MIC) was determined using a microcultivation device, and the minimal bactericidal concentration (MBC) was determined through subsequent solid medium cultivation. Additionally, the minimal biofilm inhibitory concentration (MBIC) was assessed using a polystyrene microtiter plate as biofilm carrier and measured through an assay determining the metabolic activity of biofilm cells. The results demonstrated successful synthesis of bimetallic nanoparticles using the extracts from Cannabis sativa and Vitis vinifera waste. The AgAuNPs exhibited significant antimicrobial activity against the tested Pseudomonas aeruginosa strains, inhibiting their growth and biofilm formation. These findings highlight the potential of waste valorization and circular economy in nanoparticle production and their application as effective antimicrobial agents. This study contributes to the growing field of sustainable nanotechnology and provides insights into the utilization of plant waste extracts for the synthesis of bimetallic nanoparticles with antimicrobial properties. The findings support the development of eco-friendly and cost-effective approaches for nanoparticle production while addressing the challenges of waste management and combating microbial infections.

• Publication type
• Journal Article MeSH