Phosphatase-degradable nanoparticles: A game-changing approach for the delivery of antifungal proteins

Zeynep Burcu Akkuş-Dağdeviren; Ahmad Saleh; Cristina Schöpf; Martyna Truszkowska; Doris Bratschun-Khan; Andrea Fürst; Anna Seybold; Martin Offterdinger; Florentine Marx; Andreas Bernkop-Schnürch

doi:10.1016/j.jcis.2023.05.051

Phosphatase-degradable nanoparticles: A game-changing approach for the delivery of antifungal proteins

J Colloid Interface Sci. 2023 Sep 15:646:290-300. doi: 10.1016/j.jcis.2023.05.051. Epub 2023 May 12.

Affiliations

¹ Center for Chemistry and Biomedicine, Department of Pharmaceutical Technology, Institute of Pharmacy, University of Innsbruck, Innrain 80/82, 6020 Innsbruck, Austria.
² Center for Chemistry and Biomedicine, Department of Pharmaceutical Technology, Institute of Pharmacy, University of Innsbruck, Innrain 80/82, 6020 Innsbruck, Austria; Department of Pharmacy, Universitas Mandala Waluya, A.H.Nasution, Kendari 93231, Southeast Sulawesi, Indonesia.
³ Biocenter, Institute of Molecular Biology, Medical University of Innsbruck, Innrain 80/82, 6020 Innsbruck, Austria.
⁴ Department of Zoology, University of Innsbruck, 6020 Innsbruck, Austria.
⁵ Division of Neurobiochemistry, Biooptics, Medical University of Innsbruck, Innrain 80/82, 6020 Innsbruck, Austria.
⁶ Center for Chemistry and Biomedicine, Department of Pharmaceutical Technology, Institute of Pharmacy, University of Innsbruck, Innrain 80/82, 6020 Innsbruck, Austria. Electronic address: andreas.bernkop@uibk.ac.at.

PMID: 37196502
DOI: 10.1016/j.jcis.2023.05.051

Abstract

Hypothesis: Polyphosphate nanoparticles as phosphatase-degradable carriers for Penicillium chrysogenum antifungal protein (PAF) can enhance the antifungal activity of the protein against Candida albicans biofilm.

Experiments: PAF-polyphosphate (PP) nanoparticles (PAF-PP NPs) were obtained through ionic gelation. The resulting NPs were characterized in terms of their particle size, size distribution and zeta potential. Cell viability and hemolysis studies were carried out in vitro on human foreskin fibroblasts (Hs 68 cells) and human erythrocytes, respectively. Enzymatic degradation of NPs was investigated by monitoring release of free monophosphates in the presence of isolated as well as C. albicans-derived phosphatases. In parallel, shift in zeta potential of PAF-PP NPs as a response to phosphatase stimuli was determined. Diffusion of PAF and PAF-PP NPs through C. albicans biofilm matrix was analysed by fluorescence correlation spectroscopy (FCS). Antifungal synergy was evaluated on C. albicans biofilm by determining the colony forming units (CFU).

Findings: PAF-PP NPs were obtained with a mean size of 300.9 ± 4.6 nm and a zeta potential of -11.2 ± 2.8 mV. In vitro toxicity assessments revealed that PAF-PP NPs were highly tolerable by Hs 68 cells and human erythrocytes similar to PAF. Within 24 h, 21.9 ± 0.4 μM of monophosphate was released upon incubation of PAF-PP NPs having final PAF concentration of 156 μg/ml with isolated phosphatase (2 U/ml) leading to a shift in zeta potential up to -0.7 ± 0.3 mV. This monophosphate release from PAF-PP NPs was also observed in the presence of C. albicans-derived extracellular phosphatases. The diffusivity of PAF-PP NPs within 48 h old C. albicans biofilm matrix was similar to that of PAF. PAF-PP NPs enhanced antifungal activity of PAF against C. albicans biofilm decreasing the survival of the pathogen up to 7-fold in comparison to naked PAF. In conclusion, phosphatase-degradable PAF-PP NPs hold promise as nanocarriers to augment the antifungal activity of PAF and enable its efficient delivery to C. albicans cells for the potential treatment of Candida infections.

Keywords: Antifungal protein; Biofilm; Candida albicans; Enzyme-responsive nanocarriers; Phosphatase; Polymeric nanoparticles; Polyphosphate.

MeSH terms

Antifungal Agents / chemistry
Antifungal Agents / pharmacology
Biofilms
Candida albicans
Candidiasis*
Humans
Microbial Sensitivity Tests
Nanoparticles* / chemistry
Polyphosphates

Substances

Antifungal Agents
Polyphosphates