title: A matrix reservoir for improved control of non-viral gene delivery.
abstract: Non-viral gene delivery suffers from a number of limitations including short transgene expression times and low transfection efficiency. Collagen scaffolds have previously been investigated as in vitro DNA reservoirs, which allow sustained release of genetic information. Efficient viral gene-transfer from these scaffolds has previously been demonstrated. However, due to concerns about the safety of viral gene therapy, the use of non-viral vectors may be preferable. In this study a DNA-dendrimer complex embedded in a cross-linked collagen scaffold was investigated as a reservoir for non-viral delivery. Elution from the scaffolds and transfection of seeded rat mesenchymal stem cells were used to evaluate the scaffold's ability to act as a reservoir for the complexes. Elution from the scaffolds was minimal after 2 days with a total of 25% of the complexes released after 7 days. Extended transgene expression after DNA-dendrimer complex delivery from the scaffolds in comparison to direct delivery to cells was observed. The elongated transfection period and relatively high levels of reporter gene expression are significant advantages over other non-viral gene therapy techniques. This platform has the potential to be an effective method of scaffold-mediated gene delivery suitable for in vitro and in vivo applications.
authors.affiliation: Network of Excellence in Functional Biomaterials, National Centre for Biomedical Engineering Science, National University of Ireland, Galway, Ireland.
authors.forename: Carolyn
authors.initials: C
authors.lastname: Holladay
authors.forename: Michael
authors.initials: M
authors.lastname: Keeney
authors.forename: Udo
authors.initials: U
authors.lastname: Greiser
authors.forename: Mary
authors.initials: M
authors.lastname: Murphy
authors.forename: Timothy
authors.initials: T
authors.lastname: O'Brien
authors.forename: Abhay
authors.initials: A
authors.lastname: Pandit
mesh_terms: D000818:Animals; D002460:Cell Line; D049109:Cell Proliferation; D002478:Cells, Cultured; D003094:Collagen / Q000737:chemistry*; D003432:Cross-Linking Reagents; D004247:DNA / Q000008:administration & dosage* / Q000235:genetics*; D050091:Dendrimers / Q000737:chemistry; D015870:Gene Expression; D017930:Genes, Reporter; D006801:Humans; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism; D011073:Polyamines / Q000737:chemistry; D011817:Rabbits; D051381:Rats; D013997:Time Factors; D054457:Tissue Scaffolds / Q000737:chemistry*; D014162:Transfection / Q000379:methods*
chemical_list: D003432:Cross-Linking Reagents; D050091:Dendrimers; C104700:PAMAM Starburst; D011073:Polyamines; D003094:Collagen; D004247:DNA; D008156:Luciferases