Facile Biofilm Penetration of Cationic Liposomes Loaded with DNase I/Proteinase K to Eradicate Cutibacterium acnes for Treating Cutaneous and Catheter Infections

Abstract

Background The biofilm produced by Cutibacterium acnes is a major infection threat for skin and implanted catheters. Nanoparticles provide a new approach to eradicate biofilms. The present study evaluated the capability of cationic liposomes loaded with DNase I (DNS) and proteinase K (PK) to remove preformed C. acnes biofilms. Methods DNS and PK were able to target and disassemble the biofilm by degrading extracellular polymer substances (EPS). Soyaethyl morpholinium ethosulfate (SME) was used to render a positive charge and enhance the antibacterial activity of the liposomes. Results The cationic liposomes containing enzymes yielded monodisperse nanovesicles ranging between 95 and 150 nm. The entrapment efficiency of the enzymes in the liposomes achieved a value of 67–83%. All liposomal formulations suppressed planktonic C. acnes growth at a minimum inhibitory concentration (MIC) equal to the free SME in the solution. The enzyme in the liposomal form inhibited biofilm growth much better than that in the free form, with the dual enzyme-loaded liposomes demonstrating the greatest inhibition of 54% based on a crystal violet assay. The biofilm-related virulence genes PA380 and PA1035 were downregulated by the combined enzymes in the liposomes but not the individual DNS or PK. Scanning electron microscopy (SEM) and confocal microscopy displayed reduced C. acnes aggregates and biofilm thickness by the liposomal system. The liposomes could penetrate through about 85% of the biofilm thickness. The in vitro pig skin permeation also showed a facile delivery of liposomes into the epidermis, deeper skin strata, and hair follicles. The liposomes exhibited potent activity to eliminate C. acnes colonization in mouse skin and catheters in vivo. The colony-forming units (CFUs) in the catheter treated with the liposomes were reduced by 2 logs compared to the untreated control. Conclusion The data suggested a safe application of the enzyme-loaded cationic liposomes as antibacterial and antibiofilm agents.

Bactericidal and anti-biofilm effects of uncharged and cationic ultrasound-responsive nitric oxide microbubbles on Pseudomonas aeruginosa biofilms

“…It has been shown that cationic surfactants possess innate bactericidal and anti-biofilm effects, which are exerted through electrostatic interactions with the biofilm extracellular matrix or bacterial cell membranes directly ( Zhang et al., 2017 ). Specifically, in planktonic cells, cationic particles have been shown to disrupt the integrity of bacterial cell membranes ( Fang et al., 2021 ); resulting in lysis of the cell. Whilst in biofilms, positively charged agents have been shown to disrupt the electrostatic interaction of negatively charged biofilm components, facilitating enhanced penetration of products into the biofilm and reducing overall biofilm structural integrity ( Pinto et al., 2020 ).…”

Advances in Nanotechnology for Biofilm Inhibition

“…(2021) reported that chitosan–liposome hydrogels demonstrated a high capability for eradicating S. aureus and P. aeruginosa biofilms when combined with chlorhexidine . Additionally, Fang et al (2021) observed that cationic liposomes containing DNase I/proteinase K were able to penetrate up to 85% of the biofilm matrix and exhibited the potential to eradicate Cutibacterium acnes biofilms . Moreover, Wang et al (2022) demonstrated that drug-loaded liposomes facilitated the significant degradation of S. aureus biofilms through proton-mediated burst action .…”

Plasma‐activated liquids for mitigating biofilms on food and food contact surfaces

“…Deng Li et al, 2022 Proteinases Breaking down peptide bonds within proteins causing substrates to break into shorter fragments, and eventually into amino acids. Kim & Kim, 2022;Mahdi & Hasan, 2022 DNAses Extracellular DNA disruption Fang et al, 2021;Hu et al, 2022 Biosurfactants (fengycin, viscosin, arthrofactin, surfactin, sophorolipids, and iturin) Acting as cell envelope-modifying or anti-matrix molecules. reactive species when plasma interacts with water in the gas phase (Bourke et al, 2017;2022d).…”

“…Extracellular DNA (eDNA) plays a vital role in the initial attachment and later aggregation of planktonic cells, both of which are critical steps for EPS production. Enzymes such as protease and DNase have been employed for inhibiting EPS production, both of them can be implemented alone or in combination with other sanitation strategies for achieving higher inactivation efficiency on microbial cells within biofilms (Fang et al., 2021; Kim & Kim, 2022). Studies have shown that enzymes such as proteinase K (Nguyen & Burrows, 2014), lipases (Seghal Kiran et al., 2014), and carbohydrate‐degrading enzymes (such as β‐glucans and α‐amylase) (Araújo et al., 2017) are effective in biofilm removal by attacking the main components of the biofilm matrix.…”