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Enhanced cellular uptake of albumin-based lyophilisomes when functionalized with cell-penetrating peptide TAT in HeLa cells.
- Source :
-
PloS one [PLoS One] 2014 Nov 04; Vol. 9 (11), pp. e110813. Date of Electronic Publication: 2014 Nov 04 (Print Publication: 2014). - Publication Year :
- 2014
-
Abstract
- Lyophilisomes are a novel class of biodegradable proteinaceous nano/micrometer capsules with potential use as drug delivery carrier. Cell-penetrating peptides (CPPs) including the TAT peptide have been successfully implemented for intracellular delivery of a broad variety of cargos including various nanoparticulate pharmaceutical carriers. In the present study, lyophilisomes were modified using CPPs in order to achieve enhanced cellular uptake. Lyophilisomes were prepared by a freezing, annealing, and lyophilization method and a cystein-elongated TAT peptide was conjugated to the lyophilisomes using a heterobifunctional linker. Fluorescent-activated cell sorting (FACS) was utilized to acquire a lyophilisome population with a particle diameter smaller than 1000 nm. Cultured HeLa, OVCAR-3, Caco-2 and SKOV-3 cells were exposed to unmodified lyophilisomes and TAT-conjugated lyophilisomes and examined with FACS. HeLa cells were investigated in more detail using a trypan blue quenching assay, confocal microscopy, and transmission electron microscopy. TAT-conjugation strongly increased binding and cellular uptake of lyophilisomes in a time-dependent manner in vitro, as assessed by FACS. These results were confirmed by confocal microscopy. Transmission electron microscopy indicated rapid cellular uptake of TAT-conjugated lyophilisomes via phagocytosis and/or macropinocytosis. In conclusion, TAT-peptides conjugated to albumin-based lyophilisomes are able to enhance cellular uptake of lyophilisomes in HeLa cells.
- Subjects :
- Amino Acid Sequence
Animals
Caco-2 Cells
Cattle
Cell Line, Tumor
Cell-Penetrating Peptides metabolism
Flow Cytometry
Freeze Drying
HeLa Cells
Humans
Microscopy, Confocal
Microscopy, Electron, Transmission
Particle Size
Serum Albumin, Bovine metabolism
Cell-Penetrating Peptides chemistry
Endocytosis
Serum Albumin, Bovine chemistry
Subjects
Details
- Language :
- English
- ISSN :
- 1932-6203
- Volume :
- 9
- Issue :
- 11
- Database :
- MEDLINE
- Journal :
- PloS one
- Publication Type :
- Academic Journal
- Accession number :
- 25369131
- Full Text :
- https://doi.org/10.1371/journal.pone.0110813