Protein-loaded soluble and nanoparticulate formulations of ionic polyphosphazenes and their interactions on molecular and cellular levels

被引:14
作者
Andrianov, Alexander K. [1 ]
Marin, Alexander [1 ]
Deng, Joseph [2 ]
Fuerst, Thomas R. [1 ,3 ]
机构
[1] Univ Maryland, Inst Biosci & Biotechnol Res, 9600 Gudelsky Dr, Rockville, MD 20850 USA
[2] Coll Comp Math & Nat Sci, Dept Biol, 1210 Biol Psychol Bldg,4094 Campus Dr, College Pk, MD 20742 USA
[3] Univ Maryland, Dept Cell Biol & Mol Genet, 1109 Microbiol Bldg, College Pk, MD 20742 USA
来源
MATERIALS SCIENCE AND ENGINEERING C-MATERIALS FOR BIOLOGICAL APPLICATIONS | 2020年 / 106卷
基金
美国国家科学基金会; 美国国家卫生研究院;
关键词
Self-assembly; Polymer-protein interactions; Nanoparticles; Polyphosphazene; Bacterial cells; PEGylation; NONCOVALENT PEGYLATION; DELIVERY; COMPLEXES; LYSOZYME; POLYMER; SURFACE; HYDROXYAPATITE; POLYELECTROLYTES; IMMUNOGENICITY; MICROSPHERES;
D O I
10.1016/j.msec.2019.110179
中图分类号
TB3 [工程材料学]; R318.08 [生物材料学];
学科分类号
0805 ; 080501 ; 080502 ;
摘要
Nanoparticulate and water-soluble formulations of ionic polyphosphazenes and protein cargo - lysozyme (LYZ) were prepared by their self-assembly in aqueous solutions at near physiological pH (pH 7.4) in the presence and absence of an ionic cross-linker - spermine tetrahydrochloride. Efficiency of LYZ encapsulation, physicochemical characteristics of formulations, and the effect of reaction parameters were investigated using asymmetric flow field flow fractionation (AF4) and dynamic light scattering (DLS) methods. The effect of both polymer formulations on encapsulated LYZ was evaluated using soluble oligosaccharide substrate, whereas their ability to present the protein to cellular surfaces was assessed by measuring enzymatic activity of encapsulated LYZ against Micrococcus lysodeikticus cells. It was found that both soluble and cross-linked polymer matrices reduce lysis of bacterial cells by LYZ, whereas activity of encapsulated protein against oligosaccharide substrate remained practically unchanged indicating no adverse effect of polyphosphazene on protein integrity. Moreover, nanoparticulate formulations display distinctly different behavior in cellular assays when compared to their soluble counterparts. LYZ encapsulated in polyphosphazene nanoparticles shows approximately 2.5-fold higher activity in its ability to lyse cells as compared with water-soluble LYZ-PCPP formulations. A new approach to PEGylation of polyphosphazene nanoparticles was also developed. The method utilizes a new ionic polyphosphazene derivative, which contains graft (polyethylene glycol) chains. PEGylation allows for an improved control over the size of nanoparticles and broader modulation of their cross-linking density, while still permitting for protein presentation to cellular substrates.
引用
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页数:8
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