Lyophilized Lipid Liquid Crystalline Nanoparticles as an Antimicrobial Delivery System

Lipid liquid crystalline nanoparticles (LCNPs) are unique nanocarriers that efficiently deliver antimicrobials through biological barriers. Yet, their wide application as an antimicrobial delivery system is hindered by their poor stability in aqueous dispersions. The production of dried LCNP powder via lyophilization is a promising approach to promote the stability of LCNPs. However, the impact of the process on the functionality of the loaded hydrophobic cargoes has not been reported yet. Herein, we investigated the potential of lyophilization to produce dispersible dry LCNPs loaded with a hydrophobic antimicrobial compound, gallium protoporphyrin (GaPP). The effect of lyophilization on the physicochemical characteristics and the antimicrobial activity of rehydrated GaPP-LCNPs was studied.

The rehydrated GaPP-LCNPs retained the liquid crystalline structure and were monodisperse (PDI: 0.27 ± 0.02), with no significant change in nanoparticle concentration despite the minor increase in hydrodynamic diameter (193 ± 6.5 compared to 173 ± 4.2 prior to freeze-drying). Most importantly, the efficacy of the loaded GaPP as an antimicrobial agent and a photosensitizer was not affected as similar MIC values were obtained against S. aureus (0.125 µg/mL), with a singlet oxygen quantum yield of 0.72. These findings indicate the suitability of lyophilization to produce a dry form of LCNPs and pave the way for future studies to promote the application of LCNPs as an antimicrobial delivery system.

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Materials

Glycerol monooleate (Myverol 18–92 K, Kerry ingredients, product number: 4552180, composed of 95% unsaturated monoglycerides) was kindly donated by DKSH Performance Materials Australia. Gallium protoporphyrin was purchased from Frontier Scientific (Logan, UT, USA). Trehalose, Pluronic F127, propylene glycol, uric acid and methanol with HPLC gradient grade ≥ 99.9% were purchased from Sigma Aldrich (St. Louis, MO, USA). Tryptic soy broth (TSB) and agar were purchased from Oxoid Limited; cation-adjusted Muller–Hinton broth was obtained from BD Difco™ (Thermo Fisher Scientific Australia Pty Ltd., Scoresby, VIC, Australia). All the reagents used were of analytical reagent grade, and double-distilled MQ water was used for all experiments conducted. Staphylococcus aureus Xen 29 bioluminescent strain derived from a parental strain from the American Type Culture Collection (ATCC) (part number: 119240) was kindly gifted by Prof. Allison Cowen. S.

Awad, M.; Barnes, T.J.; Prestidge, C.A. Lyophilized Lipid Liquid Crystalline Nanoparticles as an Antimicrobial Delivery System. Antibiotics 2023, 12, 1405. https://doi.org/10.3390/antibiotics12091405