The impact of agarose immobilization on the activity of lytic Pseudomonas aeruginosa phages combined with chemicals

Abstract

The implementation of non-traditional antibacterials is currently one of the most intensively explored areas of modern medical and biological sciences. One of the most promising alternative strategies to combat bacterial infections is the application of lytic phages combined with established and new antibacterials. The presented study investigates the potential of agarose-based biocomposites containing lytic Pseudomonas phages (KT28, KTN4, and LUZ19), cupric ions (Cu2+), strawberry furanone (HDMF), and gentamicin (GE) as antibacterials and anti-virulent compounds for novel wound dressings. Phages (KT28, KTN4, LUZ19, and triple-phage cocktail) alone and in combination with a triple-chemical mixture (Cu + GE + HDMF) when applied as the liquid formulation caused a significant bacterial count reduction and biofilm production inhibition of clinical P. aeruginosa strains. The immobilization in the agarose scaffold significantly impaired the bioavailability and diffusion of phage particles, depending on virion morphology and targeted receptor specificity. The antibacterial potential of chemicals was also reduced by the agarose scaffold. Moreover, the Cu + GE + HDMF mixture impaired the lytic activity of phages depending on viral particles’ susceptibility to cupric ion toxicity. Therefore, three administration types were tested and the optimal turned out to be the one separating antibacterials both physically and temporally. Taken together, the additive effect of phages combined with chemicals makes biocomposite a good solution for designing new wound dressings. Nevertheless, the phage utilization should involve an application of aqueous cocktails directly onto the wound, followed by chemicals immobilized in hydrogel dressings which allow for taking advantage of the antibacterial and anti-virulent effects of all components. Key points • The immobilization in the agarose impairs the bioavailability of phage particles and the Cu + GE + HDMF mixture. • The cupric ions are toxic to phages and are sequestrated on phage particles and agarose matrix. • The elaborated TIME-SHIFT administration effectively separates antibacterials both physically and temporally.

Chitosan-based matrix as a carrier for bacteriophages

“…The laser interferometry technique combined with the standard colony count methods showed that the number of virions released from the matrix structure was strongly limited. Similar studies conducted by Dorotkiewicz-Jach et al ( 2023 ) confirmed the immobilization of the same phages on a matrix of another polysaccharide—agarose. The lack of significant diffusion of phage particles within the chitosan matrix might be explained by the difference in net charge between the capsid (negatively charged) and the tail (positively charged) causing directional immobility in the positively charged biopolymer matrix as demonstrated by Meyer et al ( 2017 ).…”

L version of the transformed Kedem–Katchalsky equations for membrane transport of electrolyte solutions and internal energy conversion

“…2 The study of membrane transport processes in such systems is important in many areas of human activity of a cognitive and utilitarian nature, e.g., in science, technology and biomedicine. 3,4 The controlled drug release systems, membrane dressings to support the healing of chronic wounds, bioreactors for testing strategies to combat bacterial infections using lytic phage application in combination with established and novel antibacterial agents, etc., 4,5 are the main examples of biomedical applications. In these systems, the membrane is a selective barrier that ensures the separation of the phases it separates.…”

“…This role is played by polymeric membranes of various structure and composition, made of polyvinyl chloride, bacterial cellulose, graphene cellulose acetate, etc. [3][4][5][6][7][8][9] The physical quantity that characterizes non-equilibrium systems is thermodynamic entropy (S-entropy). 10,11 It plays a fundamental role in the study of non-equilibrium processes, since it numerically characterizes the degree of irreversibility of physico-chemical processes, including biological processes.…”

The Role of the Gravitational Field in Generating Electric Potentials in a Double-Membrane System for Concentration Polarization Conditions

“…The gravitational field plays an important role in membrane transport by inducing or eliminating natural (gravitational) convection [3]. The study of membrane transport processes in such systems is important in many areas of science, technology and biomedicine [4][5][6][7]. Examples of terrestrial biomedical applications include controlled drug release systems, membrane dressings to promote healing 2 of 19 of chronic wounds, bioreactors for testing strategies to combat bacterial infections using lytic phage applications in combination with established and novel antimicrobial agents, etc.…”

“…The fluid behavior for large density differences is described by the Navier-Stokes equations in the Boussinesque approximation [18][19][20][21]. For large values of R c (R c ~10 6 ), turbulent Rayleigh-Benard convection or its variants occurring on horizontal surfaces is widely studied in the context of technological applications. Turbulent natural convection also occurs in the boundary layers of the atmosphere, the oceans and other large bodies of water, as well as in the Earth's interior [21].…”