Localized delivery of DNA to the cells by viral collagen-loaded silica colloidal crystals.
Low-molecular-weight colloidal crystals with enhanced biocompatibility and ordered porous structure are used in drug-delivery systems. The objective of our study is to demonstrate the use of silica nanoscale colloid particles for localized recombinant DNA release. The colloids were coated with collagen-containing viral vector constructs of lentiviral green fluorescent protein (GFP), and solidified at 37 degrees C. The colloid-collagen-viral vector platform (CCP) was transferred to cell monolayer cultures of human lung fibroblasts. Results show specific infection of cells directly beneath the platform, as evidenced by positive GFP in their cytoplasm, while neighboring cells show no cytoplasmic GFP The infection of specific cells is probably due to the gradual release of viral particles from the collagen matrix by cell-secreted collagenase, which avoids overdosing the cells with viral particles, resulting from the cytopathic effect often seen with high-titer viral infection. Cells infected with the lentiviral-GFP or lentivirus alone, not incorporated into the colloid-collagen device, show caspase 3-associated apoptotic cell death. This suggests that colloidal crystal-coated collagen may be used as a powerful platform to deliver genes of choice to localized subgroups of specific cells of interest. This specificity in the delivery mode is beneficial for functional studies of gene-directed impact on a particular cell population of interest in a heterogeneous cell culture.
Sarojini
Harshini
H
University of Louisville School of Medicine, Louisville, KY, USA.
Medepalli
Krishnakiran
K
Terry
Derek A
DA
Alphenaar
Bruce W
BW
Wang
Eugenia
E
D002460:Cell Line; D003102:Colloids / Q000737:chemistry; D003460:Crystallization / Q000379:methods; D004247:DNA / Q000008:administration & dosage* / Q000235:genetics*; D004337:Drug Carriers / Q000737:chemistry*; D005347:Fibroblasts / Q000502:physiology*; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D012822:Silicon Dioxide / Q000737:chemistry