Sustained release of plasmid DNA using lipid microtubules and agarose hydrogel.
Non-viral gene therapy typically results in low transfection efficiencies and transient gene expression. To address these limitations, two sustained delivery systems capable of releasing functional, compacted DNA for over 50 days were designed. A luciferase plasmid was compacted with a polylysine-polyethylene glycol conjugate and released from agarose hydrogel and lipid microtubule-hydrogel delivery systems for over 50 days. The released DNA was characterized structurally using sedimentation, electron microscopy, and serum stability, and functionally using in vitro transfections. The released DNA retained its physical compaction and nuclease resistance and was converted from supercoiled to nicked and linear forms. Released compacted DNA produced significant gene expression in vitro, although at lower levels than freshly compacted DNA. Thus, hydrogels and lipid microtubules successfully provided the slow release of bioactive, compacted DNA.
Meilander
Nancy J
NJ
Biomaterials, Cell and Tissue Engineering Laboratory, Department of Biomedical Engineering, Case Western Reserve University, Wickenden Bldg. #319, 10900 Euclid Ave., Cleveland, OH 44106-7207, USA.
Pasumarthy
Murali K
MK
Kowalczyk
Tomasz H
TH
Cooper
Mark J
MJ
Bellamkonda
Ravi V
RV
D004247:DNA / Q000008:administration & dosage* / Q000097:blood / Q000235:genetics*; D004278:DNA, Superhelical / Q000008:administration & dosage / Q000097:blood / Q000737:chemistry; D003692:Delayed-Action Preparations; D018014:Gene Transfer Techniques; D006801:Humans; D020100:Hydrogels / Q000737:chemistry*; D066298:In Vitro Techniques; D008055:Lipids / Q000737:chemistry*; D008854:Microscopy, Electron; D010957:Plasmids / Q000235:genetics*; D012685:Sepharose / Q000737:chemistry*; D014162:Transfection
D004278:DNA, Superhelical; D003692:Delayed-Action Preparations; D020100:Hydrogels; D008055:Lipids; D004247:DNA; D012685:Sepharose