Fabrication and in vitro testing of polymeric delivery system for condensed DNA.
Polyethylenimine (PEI) was combined with plasmid DNA and freeze dried following the addition of sucrose as a lyoprotectant and pore-forming agent. Freeze-dried PEI DNA condensates were dry mixed with granular polylactideglycolic acid (PLGA) then compression molded and sponged to encapsulated PEI DNA. A measurement of the elastic modulus indicated that 91 wt% sucrose substituted for 95 wt% sodium chloride as a porogen, resulting in PLGA sponges with a mechanical modulus of 100 kPa. The PEI DNA was retained (80%) within PLGA sponges prepared with sucrose during the leaching and subsequent 2-week release studies, whereas sodium chloride PLGA sponges caused the premature release (100%) of PEI DNA within 2 days. In vitro gene transfer studies with PEI DNA PLGA sponges established that adherent and infiltrating fibroblasts expressed reporter gene for 15 days compared with the short, 3-day expression mediated by direct gene of PEI DNA on cells in culture. The results demonstrate an approach to encapsulate condensed DNA in a PLGA sponge for the purpose of retaining DNA within the matrices and creating efficient gene transfer during tissue engineering.
Huang
Yen-Chen
YC
College of Pharmacy, University of Michigan, Ann Arbor, Michigan 48109-1065, USA.
Connell
Maureen
M
Park
Youmie
Y
Mooney
David J
DJ
Rice
Kevin G
KG
D000818:Animals; D004270:DNA, Circular / Q000737:chemistry* / Q000378:metabolism; D004548:Elasticity; D005347:Fibroblasts / Q000166:cytology / Q000502:physiology; D005786:Gene Expression Regulation; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D019344:Lactic Acid / Q000737:chemistry*; D008422:Materials Testing; D051379:Mice; D041681:NIH 3T3 Cells; D010316:Particle Size; D010957:Plasmids / Q000737:chemistry* / Q000378:metabolism; D011094:Polyethyleneimine / Q000737:chemistry*; D011100:Polyglycolic Acid / Q000737:chemistry*; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011108:Polymers / Q000737:chemistry*; D013395:Sucrose / Q000737:chemistry; D013499:Surface Properties
D004270:DNA, Circular; D011108:Polymers; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011100:Polyglycolic Acid; D019344:Lactic Acid; D013395:Sucrose; D011094:Polyethyleneimine