Linked Life Data

15761942

Source:http://linkedlifedata.com/resource/pubmed/id/15761942

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
1
pubmed:dateCreated
2005-3-11
pubmed:abstractText
In order to develop a systemically administered safe and effective nonviral gene delivery system, cellular interactions and plasmid DNA transfection with poly(ethylene glycol)-modified (PEGylated) gelatin nanoparticles were examined. The DNA-containing nanoparticles were prepared by a controlled water-ethanol solvent displacement method. The nanoparticles were characterized for particle size, surface charge, and DNA loading, release, and stability. For cellular interaction studies, the control and PEGylated gelatin nanoparticles, complexed either with colloidal gold for transmission electron microscopy or loaded with rhodamine-dextran for fluorescence confocal microscopy, were incubated with NIH-3T3 fibroblast cells. At different time points, the location of the nanoparticles in the cellular environment was investigated. Furthermore, a reporter plasmid expressing the enhanced green fluorescent protein was encapsulated in the control gelatin and PEGylated gelatin nanoparticles for in vitro transfection studies. DNA-containing nanoparticles were prepared in the size range of 100-500 nm, with an average of 200 nm. PEGylated gelatin nanoparticles, with a slight negative surface charge, could stably and efficiently encapsulate plasmid DNA. Both transmission electron microscopy and confocal microscopy images showed that the gelatin and PEGylated gelatin nanoparticles rapidly entered the cell through nonspecific endocytosis followed by vesicular transport in the cytosol. Almost 100% of the administered gelatin and PEGylated gelatin nanoparticles were internalized in NIH-3T3 cells within the first 6 h of incubation. A large fraction of the administered nanoparticles was found to be concentrated in the perinuclear region of the cells after 12 h. Green fluorescent protein expression was observed after 12 h of nanoparticle incubation and remained stable for up to 96 h. Flow cytometry results showed that the DNA transfection efficiency with gelatin and PEGylated gelatin nanoparticles was 43% and 61%, respectively, after 96 h. The results of this study illustrate that PEGylated gelatin nanoparticles were rapidly internalized by the cells through nonspecific endocytosis and remained intact in the cytosol for up to 12 h. In addition, the DNA-encapsulated PEGylated gelatin nanoparticles were found to efficiently transfect cells in culture.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Jan
pubmed:issn
0022-3549
pubmed:author
pubmed:issnType
Print
pubmed:volume
94
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
184-98
pubmed:dateRevised
2007-11-14
pubmed:meshHeading
pubmed:year
2005
pubmed:articleTitle
Cellular interactions and in vitro DNA transfection studies with poly(ethylene glycol)-modified gelatin nanoparticles.
pubmed:affiliation
Department of Pharmaceutical Sciences, School of Pharmacy, Northeastern University, 110 Mugar Life Sciences Building, Boston, Massachusetts 02115, USA.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S.