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Effect of Cholesterol on the Properties of Spray-Dried Lysozyme-Loaded Liposomal Powders

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Abstract

The influence of cholesterol (Chol) in the liposomal bilayer on the properties of inhalable protein-loaded liposomal powders prepared by spray-drying technique was investigated. Lysozyme (LSZ) was used as a model protein. Feed solution for spray drying was prepared by direct mixing of aqueous solution of LSZ with mannitol solution and empty liposome dispersions composed of hydrogenated phosphatidylcholine and Chol at various molar ratios. The spray-dried powders were characterized with respect to morphology, thermal property, and crystallinity using scanning electron microscopy, differential scanning calorimetry, and X-ray diffraction, respectively. Most formulations gave slightly aggregated, spherical particles, and percentage yields of the spray-dried powders decreased with increasing Chol content. Degree of particle aggregation depended on the powder composition. The powders spontaneously formed liposomes which efficiently entrapped LSZ after reconstitution with HEPES buffered saline (HBS) at 37°C. Lysozyme entrapment efficiency and size distribution of the reconstituted liposomes were evaluated after the powders were reconstituted with HBS. Increasing Chol content resulted in a decrease in size of the reconstituted liposomes and an increase in entrapment efficiency of LSZ. These results correlated with thermal behaviors of the reconstituted liposomes. Biological activity of LSZ was not affected by the spray-drying process. It was also demonstrated that LSZ-loaded liposomal powders could be produced without the need to preload the LSZ into liposomes prior to spray-drying process.

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Abbreviations

Chol:

Cholesterol

DPPC:

Dipalmitoylphosphatidylcholine

DRV:

Dehydrated-rehydrated vesicles

DSC:

Differential scanning calorimetry

EE:

Entrapment efficiency

HBS:

HEPES buffered saline

HPC:

Hydrogenated soybean phosphatidylcholine

LSZ:

Lysozyme

M:

Mannitol

MMD:

Mass median diameter

PC:

Phosphatidylcholine

QPBCA:

QuantiPro bicinchoninic acid

SEM:

Scanning electron microscopy

T d :

Denaturation melting temperature

T m :

Phase transition temperature

XRPD:

X-ray powder diffraction

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Acknowledgments

The authors are thankful to grants from the Graduate School of Chulalongkorn University, Faculty of Pharmaceutical Sciences and from the Commission on Higher Education of Thailand.

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  1. Department of Pharmaceutics and Industrial Pharmacy, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok, 10330, Thailand

    Dusadee Charnvanich, Nontima Vardhanabhuti & Poj Kulvanich

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Correspondence to Poj Kulvanich.

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Charnvanich, D., Vardhanabhuti, N. & Kulvanich, P. Effect of Cholesterol on the Properties of Spray-Dried Lysozyme-Loaded Liposomal Powders. AAPS PharmSciTech 11, 832–842 (2010). https://doi.org/10.1208/s12249-010-9442-8

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