Delivery of non-viral gene carriers from sphere-templated fibrin scaffolds for sustained transgene expression.
Delivery of safe and controlled levels of biomimetic cues to govern host response and reorganization is a fundamental component in the design of tissue engineering scaffolds. Non-viral gene delivery is an approach that exploits the cell machinery to produce proteins while avoiding genomic DNA incorporation. We describe a method to integrate polymeric non-viral gene carriers (polyplexes) within a novel three-dimensional, sphere-templated fibrin scaffold suitable for soft tissue engineering applications. After seeding the scaffolds with NIH-3T3 fibroblasts, different transgene expression profiles were achieved based on the spatial distribution of polyplexes within the scaffold. Scaffolds with polyplexes coated onto the surface of inter-connected pores showed peak transfection at day 5 and linear expression through 15 days. Scaffolds with polyplexes embedded within the fibrils of the biopolymer showed peak expression at 7-9 days and showed linear expression for 21-29 days, depending on the polymer:DNA ratio. Surface-coated polyplexes achieved one order of magnitude greater expression than polyplexes embedded within the scaffold. The integrated material formulations developed in this work provide a useful technology for tissue engineering applications by demonstrating the ability to provide long-term biomimetic cues through non-viral gene delivery.
Department of Bioengineering, University of Washington, 1705 NE Pacific Street, W.H. Foege Building, Room N530P, Mailstop 355061, Seattle, WA 98195, USA.
Justin M
JM
Saul
Michael P
MP
Linnes
Buddy D
BD
Ratner
Cecilia M
CM
Giachelli
Suzie H
SH
Pun
D016475:3T3 Cells; D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry; D004247:DNA / Q000008:administration & dosage* / Q000737:chemistry*; D004337:Drug Carriers / Q000737:chemistry*; D005337:Fibrin / Q000737:chemistry*; D015870:Gene Expression / Q000502:physiology*; D008422:Materials Testing; D051379:Mice; D014162:Transfection / Q000379:methods*
D001672:Biocompatible Materials; D004337:Drug Carriers; D005337:Fibrin; D004247:DNA