Plasmid delivery in vivo from porous tissue-engineering scaffolds: transgene expression and cellular transfection.
Tissue engineering scaffolds capable of sustained plasmid release can promote gene transfer locally and stimulate new tissue formation. We have investigated the scaffold design parameters that influence the extent and duration of transgene expression and have characterized the distribution of transfected cells. Porous scaffolds with encapsulated plasmid were fabricated from poly(lactide-co-glycolide) with a gas foaming procedure, with wet granulation employed to mix the components homogeneously prior to foaming. Wet granulation enhanced plasmid incorporation relative to standard procedures and also enhanced in vivo transgene expression, possibly through the increased loading and maintenance of the scaffold pore structure. The plasmid loading regulated the quantity and duration of transgene expression, with expression for 105 days achieved at the highest dosage. Expression was localized to the implantation site, though the distribution of transfected cells varied with time. Transfected cells were initially observed at the scaffold periphery (day 3), then within the pores and adjacent to the polymer (day 17), and finally throughout the scaffold interior (day 126). Delivery of a plasmid encoding VEGF increased the blood vessel density relative to control. Correlating scaffold design with gene transfer efficiency and tissue formation will facilitate application of plasmid-releasing scaffolds to multiple tissues.
Jang
Jae-Hyung
JH
Department of Chemical and Biological Engineering, Northwestern University, 2145 Sheridan Road E156, Evanston, IL 60208-3120, USA.
Rives
Christopher B
CB
Shea
Lonnie D
LD
D000818:Animals; D001672:Biocompatible Materials; D004247:DNA / Q000378:metabolism; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics*; D007150:Immunohistochemistry; D008156:Luciferases / Q000378:metabolism; D008297:Male; D051379:Mice; D009389:Neovascularization, Pathologic; D010957:Plasmids / Q000378:metabolism; D013997:Time Factors; D023822:Tissue Engineering / Q000379:methods*; D014162:Transfection; D019076:Transgenes
D001672:Biocompatible Materials; D004247:DNA; D008156:Luciferases