Roles of chitosan in synthesis, antibacterial and anti-biofilm properties of bionano silver and gold.

Antibiotic-resistance and bacterial bioburden on wound surfaces are the significant challenges to wound healing. Silver and gold nanoparticles (are termed as AgNPs and AuNPs) have been investigated as alternative antimicrobial agents to combat antibiotic-resistant bacterial infections owing to their antibacterial and anti-biofilm activities. Chitosan (CS) has largely been used in nanoparticle synthesis as a stabilizing or capping agent. In this study, AgNPs and AuNPs were synthesized using different concentrations of aqueous extract of tiger milk mushroom (Lignosus rhinocerotis) (WETMM) and CS as reducing and stabilizing agent, respectively. Particle size and morphology of both were determined by dynamic light scattering (DLS) method and transmission electron microscopic analysis (TEM). FTIR analysis was conducted to determine the interactions between nanoparticle precursors. The observed peaks at 450 nm and 534-565 nm using a spectrophotometer were corresponded to the surface Plasmon resonance of AgNPs and AuNPs respectively, indicating the formation of respective nanoparticles. FTIR analysis confirmed the role of WETMM as a reducing agent and CS as a stabilizer of AgNPs and AuNPs. Faster formation of nanoparticles was observed besides an increase in particle size when higher CS concentrations were used. TEM micrographs revealed the spherical shape of most nanoparticles with particle sizes in the range of 4 to 58 nm and 18 to 28 nm for AgNPs and AuNPs, respectively. Both nanoparticles exhibited antimicrobial activity against Gram-positive and -negative bacteria, with AgNPs showing a superior antibacterial efficacy than AuNPs. Both microbroth dilution and agar well diffusion assays indicated that CS was an important component to facilitate antibacterial activity for AuNPs. Contrarily, CS stabilization did not enhance the antibacterial efficacy of AgNPs. CS-stabilized AgNPs and AuNPs achieved biofilm inhibition of 53.21% and 79.39% for Pseudomonas aeruginosa and 48.71% and 48.16% for Staphylococcus aureus, respectively. Similarly, CS stabilization enhanced the anti-biofilm activity of AuNPs but no such effect was seen for AgNPs. In conclusion, CS-stabilized AgNPs and AuNPs possess both antimicrobial and anti-biofilm activities. However, CS acted differently when combined with AgNPs and AuNPs, needing further investigation and optimization to improve the antimicrobial activity of both nanoparticles.

Gelatin Hydrogels Loaded with Lactoferrin-Functionalized Bio-Nanosilver as a Potential Antibacterial and Anti-Biofilm Dressing for Infected Wounds: Synthesis, Characterization, and Deciphering of Cytotoxicity.

Gelatin hydrogels are attractive for wound applications owing to their well-defined structural, physical, and chemical properties as well as good cell adhesion and biocompatibility. This study aimed to develop gelatin hydrogels incorporated with bio-nanosilver functionalized with lactoferrin (Ag-LTF) as a dual-antimicrobial action dressing, to be used in treating infected wounds. The hydrogels were cross-linked using genipin prior to loading with Ag-LTF and characterized for their physical and swelling properties, rheology, polymer and actives interactions, and in vitro release of the actives. The hydrogel's anti-biofilm and antibacterial performances against S. aureus and P. aeruginosa as well as their cytotoxicity effects were assessed in vitro, including primary wound healing gene expression of human dermal fibroblasts (HDFs). The formulated hydrogels showed adequate release of AgNPs and LTF, with promising antimicrobial effects against both bacterial strains. The Ag-LTF-loaded hydrogel did not significantly interfere with the normal cellular functions as no alteration was detected for cell viability, migration rate, and expression of the target genes, suggesting the nontoxicity of Ag-LTF as well as the hydrogels. In conclusion, Ag-LTF-loaded genipin-cross-linked gelatin hydrogel was successfully synthesized as a new approach for fighting biofilms in infected wounds, which may be applied to accelerate healing of chronic wounds.

Antimicrobial activity of multifaceted lactoferrin or graphene oxide functionalized silver nanocomposites biosynthesized using mushroom waste and chitosan.

Hybrid nanoparticles designed to exert multiple mechanisms of antibacterial action offer a new approach to the fight against pathogenic resistant bacteria. In this study, nanomaterials with the dual actions of antibacterial and anti-biofilm activities were developed using silver nanoparticles (AgNPs) functionalized with either lactoferrin (LTF) or graphene oxide (GO). AgNPs were synthesized using mushroom waste as a reducing agent and chitosan (CS) as a stabilizing agent, prior to their surface functionalization with either GO (AgGO) or LTF (Ag-LTF). The AgNPs exhibited a surface plasmon resonance (SPR) band at 430 nm, as determined by UV-vis spectroscopy, whereas the absorption of AgGO and Ag-LTF occurred at 402 and 441 nm, respectively. Particle size analysis of AgNPs, AgGO, and Ag-LTF revealed sizes of 121.5 ± 10.5, 354.0 ± 1.6, and 130.8 ± 1.2 nm, respectively. All AgNPs, Ag-LTF, and AgGO inhibited selected Gram-positive bacteria and Gram-negative bacteria with comparable antibacterial performance, as determined by the agar diffusion method. Despite the absence of antibacterial activity by GO and LTF, a synergistic effect of AgGO and Ag-LTF was observed as they had a greater activity against P. aeruginosa. Moreover, Ag-LTF did not affect cell viability and migration rate of cells, suggesting the non-toxicity of Ag-LTF. In conclusion, AgNPs, Ag-LTF, and AgGO possess antibacterial activity, which may offer an alternative for future antibacterial agents.