Targeted Propolis-Loaded Poly (Butyl) Cyanoacrylate Nanoparticles: An Alternative Drug Delivery Tool for the Treatment of Cryptococcal Meningitis

Patcharin Thammasit; Chayada Sitthidet Tharinjaroen; Yingmanee Tragoolpua; Volker Rickerts; Radostina Georgieva; Hans Bäumler; Khajornsak Tragoolpua

doi:10.3389/fphar.2021.723727

Targeted Propolis-Loaded Poly (Butyl) Cyanoacrylate Nanoparticles: An Alternative Drug Delivery Tool for the Treatment of Cryptococcal Meningitis

Front Pharmacol. 2021 Aug 20:12:723727. doi: 10.3389/fphar.2021.723727. eCollection 2021.

Authors

Patcharin Thammasit^{1

2

3}, Chayada Sitthidet Tharinjaroen^{1

4}, Yingmanee Tragoolpua⁵, Volker Rickerts⁶, Radostina Georgieva^{2

7}, Hans Bäumler², Khajornsak Tragoolpua^{1

4}

Affiliations

¹ Division of Clinical Microbiology, Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai, Thailand.
² Charité-Universitätsmedizin Berlin, Institute of Transfusion Medicine, Berlin, Germany.
³ Department of Microbiology, Faculty of Medicine, Chiang Mai University, Chiang Mai, Thailand.
⁴ Infectious Disease Research Unit (IDRU), Division of Clinical Microbiology, Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai, Thailand.
⁵ Department of Biology, Faculty of Science, Chiang Mai University, Chiang Mai, Thailand.
⁶ Mycotic and Parasitic Agents and Mycobacteria, Department of Infectious Diseases, Robert Koch Institute, Berlin, Germany.
⁷ Department of Medical Physics, Biophysics and Radiology, Medical Faculty, Trakia University, Stara Zagora, Bulgaria.

Abstract

In this study, we describe a nano-carrier system for propolis that is able to cross an in vitro model of the blood-brain barrier (BBB) and effectively reduce the virulence of Cryptococcus neoformans in animal models. Antimicrobial properties of propolis have been widely studied. However, propolis applications are limited by its low water solubility and poor bioavailability. Therefore, we recently formulated novel poly (n-butyl cyanoacrylate) nanoparticles (PBCA-NP) containing propolis. PBCA-NP are biocompatible, biodegradable and have been shown to effectively cross the BBB using apolipoprotein E (ApoE) as a ligand. Prepared nanoparticles were characterized for particle size, zeta potential, propolis entrapment efficiency and in vitro release. Additionally, the PBCA-NP were functionalized with polysorbate 80, which then specifically adsorbs ApoE. Using an in vitro BBB model of human brain microvascular endothelial cells hCMEC/D3, it was shown that fluorescence labelled ApoE-functionalized PBCA-NP were internalized by the cells and translocated across the cell monolayer. Propolis-loaded PBCA-NP had in vitro, antifungal activity against C. neoformans, which causes meningitis. To utilize the invertebrate model, Galleria mellonella larvae were infected with C. neoformans and treated with propolis-loaded PBCA-NP. The larvae exhibited normal behavior in toxicity testing, and treatment with propolis-loaded PBCA-NP increased survival in the C. neoformans-infected larvae group. In addition, following cryptococcal infection and then 7 days of treatment, the tissue fungal burden of mice treated with propolis-loaded PBCA-NP was significantly lower than control groups. Therefore, our ApoE-functionalized propolis-loaded PBCA-NP can be deemed as a potential targeted nanoparticle in the therapeutic treatment of cerebral cryptococcosis.

Keywords: Cryptococcus neoformans; blood-brain barrier; in vivo model; poly (n-butyl cyanoacrylate) nanoparticles; propolis.