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Thermosensitive hydrogel for prolonged delivery of lentiviral vector expressing neurotrophin‐3 in vitro
Author(s) -
McMahon Siobhan S,
Nikolskaya Natalia,
Choileáin Siobhan Ní,
Hennessy Niamh,
O'Brien Timothy,
Strappe Padraig M,
Gorelov Alexander,
Rochev Yury
Publication year - 2011
Publication title -
the journal of gene medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.689
H-Index - 91
eISSN - 1521-2254
pISSN - 1099-498X
DOI - 10.1002/jgm.1613
Subject(s) - chitosan , viral vector , gene delivery , green fluorescent protein , chemistry , cell sorting , neurite , in vitro , microbiology and biotechnology , neurotrophin 3 , genetic enhancement , biochemistry , biology , recombinant dna , gene , neurotrophic factors , brain derived neurotrophic factor , receptor
Background The development of tissue engineering scaffolds for gene delivery has the potential to enhance gene transfer efficiency and safety via controlled temporal and spatial delivery. Lentiviral delivery can be carried out using the natural biopolymer thermoresponsive gel, chitosan/β‐glycerol phosphate (β‐GP) as a carrier. Methods Three chitosan/β‐GP scaffolds were prepared with varying concentrations of chitosan and β‐GP to obtain a pH and gelation temperature suitable for in situ delivery. A lentiviral vector expressing either green fluorescent protein (Lenti GFP) or neurotrophin‐3 (Lenti NT‐3) was incorporated into the chitosan/β‐GP scaffolds and also into collagen 0.1% w/v (control). Viral elution medium was removed at various timepoints and added to the culture medium of pre‐seeded HeLa or primary dorsal root ganglia (DRG) cells, respectively. GFP gene expression was quantified using fluorescence‐activated cell sorting analysis. The effect of Lenti NT‐3 was analyzed by measuring DRG neurite outgrowth. Results Collagen displayed its most significant elution of virus on day 1 and chitosan/β‐GP (with a final concentration of 2.17% chitosan) on day 3. Conclusions The system shows promise for the in situ , thermoresponsive delivery of lentiviral vectors providing long‐term gene expression for therapeutic factors to treat conditions such as injury to the nervous system. Copyright © 2011 John Wiley & Sons, Ltd.