The impact of surface charge on the interaction of cholesterol-free fusogenic liposomes with planktonic microbial cells and biofilms

Abstract

This study focused on the development of cholesterol-free fusogenic liposomes with different surface charge with the aim of improving biofilm penetration. In vitro assessments of the liposomes included physical stability, biocompatibility, fusion with microbial cells, and the ability to penetrate established biofilms. Using dynamic light scattering, cholesterol-free, fusogenic liposomes were found to be < 200 nm in size with small size distribution (PDI < 0.1) and physically stable for a year when stored at 4◦C. Transmission electron microscopy (TEM) images confirmed vesicular sizes for selected liposomal formulations. Liposomal ability to fuse with microbial cells was assessed using lipid mixing and flow cytometer assays. Fusion levels were found to be higher with Escherichia coli compared to Staphylococcus aureus and Candida albicans regardless of the liposomal charge. Neutral liposomes exhibited highest fusion, followed by cationic and anionic liposomes, respectively. Our investigations demonstrated that fusion is a multifactorial process influenced by the chemical composition of the liposomes, the liposomal surface charge, and components of the microbial cell envelope. Penetration and retention within preformed S. aureus biofilms were assessed for liposomes with various surface charges. All liposomes, regardless of surface charge, were capable of penetrating and diffusing through the biofilm matrix. However, cationic liposomes displayed greatest interaction and retention. Biocompatibility was confirmed through haemolysis and cytotoxicity studies. The cholesterol-free fusogenic liposomes developed in this study demonstrated promising potential as drug delivery systems for incorporating antimicrobial agents for biofilm treatment.