Journal article
Long-term in vivo gene expression via delivery of PEI-DNA condensates from porous polymer scaffolds
Human gene therapy, Vol.16(5), pp.609-617
2005
DOI: 10.1089/hum.2005.16.609
PMID: 15916485
Abstract
Nonviral delivery vectors are attractive for gene therapy approaches in tissue engineering, but suffer from low transfection efficiency and short-term gene expression. We hypothesized that the sustained delivery of poly(ethylenimine) (PEI)-condensed DNA from three-dimensional biodegradable scaffolds that encourage cell infiltration could greatly enhance gene expression. To test this hypothesis, a PEI-condensed plasmid encoding beta-galactosidase was incorporated into porous poly(lactide-co-glycolide) (PLG) scaffolds, using a gas foaming process. Four conditions were examined: condensed DNA and uncondensed DNA encapsulated into PLG scaffolds, blank scaffolds, and bolus delivery of condensed DNA in combination with implantation of PLG scaffolds. Implantation of scaffolds incorporating condensed beta-galactosidase plasmid into the subcutaneous tissue of rats resulted in a high level of gene expression for the entire 15-week duration of the experiment, as exemplified by extensive positive staining for beta-galactosidase gene expression observed on the exterior surface and throughout the cross-sections of the explanted scaffolds. No positive staining could be observed for the control conditions either on the exterior surface or in the cross-section at 8- and 15-week time points. In addition, a high percentage (55-60%) of cells within scaffolds incorporating condensed DNA at 15 weeks demonstrated expression of the DNA, confirming the sustained uptake and expression of the encapsulated plasmid DNA. Quantitative analysis of beta-galactosidase gene expression revealed that expression levels in scaffolds incorporating condensed DNA were one order of magnitude higher than those of other conditions at the 2- week time point and nearly two orders of magnitude higher than those of the control conditions at the 8- and 15-week time points. This study demonstrated that the sustained delivery of PEI-condensed plasmid DNA from PLG scaffolds led to an in vivo long-term and high level of gene expression, and this system may find application in areas such as bone tissue engineering.
Details
- Title: Subtitle
- Long-term in vivo gene expression via delivery of PEI-DNA condensates from porous polymer scaffolds
- Creators
- Yen-Chen HUANG - Department of Biomedical Engineering, University of Michigan, Ann Arbor, MI 48109, United StatesKathryn RIDDLE - Department of Chemical Engineering, University of Michigan, Ann Arbor, MI 48109, United StatesKevin G RICE - College of Pharmacy, University of Iowa, Iowa City, IA 52242, United StatesDavid J MOONEY - Division of Engineering and Applied Sciences, Harvard University, Cambridge, MA 02138, United States
- Resource Type
- Journal article
- Publication Details
- Human gene therapy, Vol.16(5), pp.609-617
- DOI
- 10.1089/hum.2005.16.609
- PMID
- 15916485
- NLM abbreviation
- Hum Gene Ther
- ISSN
- 1043-0342
- eISSN
- 1557-7422
- Publisher
- Liebert; Larchmont, NY
- Language
- English
- Date published
- 2005
- Academic Unit
- Pharmaceutical Sciences and Experimental Therapeutics; Craniofacial Anomalies Research Center; Nephrology; Medicinal and Natural Products Chemistry; Internal Medicine
- Record Identifier
- 9984065691302771
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