Targeted cytosolic delivery of hydrogel nanoparticles into HepG2 cells through engineered Sendai viral envelopes

FEBS Letters

Gupta, Chhitar M¹  Jana, Siddhartha S³  Maitra, Amarnath²  Sarkar, Debi P³  Mani, Prashant³  Bharali, Dhruba J² 

[1] Division of Molecular and Structural Biology, Central Drug Research Institute, P.O. Box 173, Lucknow 226001, India;Department of Chemistry, University of Delhi, Delhi 110007, India;Department of Biochemistry, University of Delhi South Campus, Benito Juarez Road, New Delhi 110021, India

关键词: Hydrogel nanoparticle;    Sendai virus;    Virosome;    Drug delivery;    Targeted cytosolic delivery;    Controlled release;    ASGP-R;    asialoglycoprotein receptor;    CHO;    Chinese hamster ovary;    DMEM;    Dulbecco's modified Eagle's medium;    DPBS;    Dulbecco's phosphate-buffered saline;    EDTA;    ethylenediaminetetraacetate (disodium salt);    F;    fusion protein;    FCS;    fetal calf serum;    FITC-Dx;    fluorescein isothiocyanate dextran (mol wt. 19.3 kDa);    PL;    polylysine;    PVPf-NP;    FITC-Dx-loaded polyvinylpyrrolidone nanoparticle;    TBS;    Tris-buffered saline;   

DOI  :  10.1016/S0014-5793(02)02467-5

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Hydrogel nanoparticles of cross-linked polyvinylpyrrolidone (PVP-NP) (35–50 nm in diameter) containing fluoresceinated dextran (FITC-Dx) were encapsulated in reconstituted Sendai viral envelopes containing only the fusion (F) protein (F-virosomes¹). Incubation of these loaded F-virosomes with human hepatoblastoma cells (HepG2) in culture resulted in membrane-fusion-mediated delivery of NPs to the cell cytoplasm, as inferred from the ability of cells to internalize FITC-Dx loaded PVP-NP (PVP^f-NP) in the presence of azide (an inhibitor of the endocytotic process). Introduction of PVP^f-NP into the HepG2 cells was assured by selective accumulation of FITC fluorescence in the cytosolic compartment. The structural integrity of the internalized PVP^f-NP was also confirmed by fluorescence microscopy and ultracentrifugation analysis. The potential usefulness of PVP-NP-mediated cytosolic release of water soluble drugs both in vitro and in vivo has been established for the first time.

【 授权许可】

Unknown