Generation of Hybrid Peptide-Silver Nanoparticles for Antibacterial and Antifouling Applications

Abstract

An alarming increase of antibiotic-resistant bacterial strains has made the demand for novel antibacterial agents, for example, more effective antibiotics, highly crucial. One of the oldest antimicrobial agents is elementary silver which has been used for thousands of years. Even in our days, elementary silver is used for medical purposes, such as for burns, wounds, and microbial infections. We have taken the effectiveness of elementary silver into consideration to generate novel antibacterial and antifouling agents. Our innovative antibacterial agents are hybrid peptide silver nanoparticles (CH-01-AgNPs) that are created de novo and in situ from a silver nitrate solution (AgNO3) in the presence of ultrashort self-assembling peptides compounds. The nucleation of CH-01-AgNPs is initiated by irradiating the peptide solution mixed with the AgNO3 solution using ultraviolet (UV) light at a wavelength of 254 nm, in the absence of any reducing or capping agents. Obviously, the peptide itself serves as the reducing agent. The ultrashort peptides are four amino acids in length with an innate ability to self-assemble into nanofibrous scaffolds. Using these ultrashort peptides CH-01 we were able to create hybrid peptide silver nanoparticles CH-01-AgNPs with a diameter of 4-6 nm. The synthesized CH-01-AgNPs were further characterized using ultraviolet-visible spectroscopy, transmission electron microscopy, dynamic light scattering, and X-ray photoelectron spectroscopy. The antibacterial and antifouling activity of CH-01-AgNPs were then investigated using either gram-negative bacteria, such as antibiotic-resistant Top10 Escherichia coli and Pseudomonas aeruginosa PDO300, or gram-positive bacteria, such as Staphylococcus aureus CECT 976. The hybrid nanoparticles demonstrated very promising antibacterial and antifouling activity with higher antibacterial and antifouling activity as commercial silver nanoparticles. Quantitative Polymerase Chain Reaction (qPCR) results showed upregulation of stress-related genes, e.g. osmB and bdm. Biocompatibility studies of CH-01-AgNPs, using concentrations of 0.06 mM and 0.125 mM, testing for the viability of human dermal fibroblast neonatal (HDFn) cells, showed no significant influence on cell viability. In summary, we consider hybrid peptide silver nanoparticles CH-01-AgNPs as promising biomaterials that can be utilized in various biomedical applications, in particular for wound healing and biofilm inhibition, but also for other applications, such as tissue engineering, drug delivery, regenerative medicine, and biosensing.