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In Vitro Characterization of Inhalable Cationic Hybrid Nanoparticles as Potential Vaccine Carriers

Research Authors
Iman M. Alfagih, Kan Kaneko, Nitesh K. Kunda, Fars Alanazi, Sarah R. Dennison, Hesham M. Tawfeek and Imran Y. Saleem
Research Department
Research Journal
Pharmaceuticals
Research Publisher
MDPI
Research Rank
1
Research Vol
14 (164)
Research Website
https://doi.org/10.3390/ph14020164
Research Year
2021
Research Abstract

In this study, PGA-co-PDL nanoparticles (NPs) encapsulating model antigen, bovine
serum albumin (BSA), were prepared via double emulsion solvent evaporation. In addition, chitosan
hydrochloride (CHL) was incorporated into the external phase of the emulsion solvent method, which
resulted in surface adsorption onto the NPs to form hybrid cationic CHL NPs. The BSA encapsulated
CHL NPs were encompassed into nanocomposite microcarriers (NCMPs) composed of L-leucine to
produce CHL NPs/NCMPs via spray drying. The CHL NPs/NCMPs were investigated for in vitro
aerosolization, release study, cell viability and uptake, and stability of protein structure. Hybrid
cationic CHL NPs (CHL: 10 mg/mL) of particle size (480.2 ± 32.2 nm), charge (+14.2 ± 0.72 mV), and
BSA loading (7.28 ± 1.3 µg/mg) were produced. The adsorption pattern was determined to follow
the Freundlich model. Aerosolization of CHL NPs/NCMPs indicated fine particle fraction (FPF:
46.79 ± 11.21%) and mass median aerodynamic diameter (MMAD: 1.49 ± 0.29 µm). The BSA α-helical
structure was maintained, after release from the CHL NPs/NCMPs, as indicated by circular dichroism.
Furthermore, dendritic cells (DCs) and A549 cells showed good viability (≥70% at 2.5 mg/mL after
4–24 h exposure, respectively). Confocal microscopy and flow cytometry data showed hybrid cationic
CHL NPs were successfully taken up by DCs within 1 h of incubation. The upregulation of CD40,
CD86, and MHC-II cell surface markers indicated that the DCs were successfully activated by the
hybrid cationic CHL NPs. These results suggest that the CHL NPs/NCMPs technology platform
could potentially be used for the delivery of proteins to the lungs for immunostimulatory applications
such as vaccines.