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Spontaneous gene transfection of human bone cells using 3D mineralized alginate–chitosan macrocapsules
Author(s) -
Green David W.,
Kim EunJung,
Jung HanSung
Publication year - 2015
Publication title -
journal of biomedical materials research part a
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.849
H-Index - 150
eISSN - 1552-4965
pISSN - 1549-3296
DOI - 10.1002/jbm.a.35414
Subject(s) - transfection , gene delivery , genetic enhancement , stromal cell , chitosan , cell culture , bone marrow , microbiology and biotechnology , reporter gene , materials science , biology , gene , cancer research , biochemistry , immunology , gene expression , genetics
The effectiveness of nonviral gene therapy remains uncertain because of low transfection efficiencies and high toxicities compared with viral‐based strategies. We describe a simple system for transient transfection of continuous human cell lines, with low toxicity, using mineral‐coated chitosan and alginate capsules. As proof‐of‐concept, we demonstrate transfection of Saos‐2 and MG63 human osteosarcoma continuous cell lines with gfp, LacZ reporter genes, and a Sox‐9 carrying plasmid, to illustrate expression of a functional gene with therapeutic relevance. We show that continuous cell lines transfect with significant efficiency of up to 65% possibly through the interplay between chitosan and DNA complexation and calcium/phosphate‐induced translocation into cells entrapped within the 3D polysaccharide based environment, as evidenced by an absence of transfection in unmineralized and chitosan‐free capsules. We demonstrated that our transfection system was equally effective at transfection of primary human bone marrow stromal cells. To illustrate, the Sox‐9, DNA plasmid was spontaneously expressed in primary human bone marrow stromal cells at 7 days with up to 90% efficiency in two repeats. Mineralized polysaccharide macrocapsules are gene delivery vehicles with a number of biological and practical advantages. They are highly efficient at self‐transfecting primary bone cells, with programmable spatial and temporal delivery prospects, premineralized bone‐like environments, and have no cytotoxic effects, as compared with many other nonviral systems. © 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 103A: 2855–2863, 2015.