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Spontaneous gene transfection of human bone cells using 3D mineralized alginate-chitosan macrocapsules

Authors
 David W. Green ; Eun-Jung Kim ; Han-Sung Jung 
Citation
 Journal of Biomedical Materials Research Part A, Vol.103(9) : 2855~2863, 2015 
Journal Title
 Journal of Biomedical Materials Research Part A 
ISSN
 1549-3296 
Issue Date
2015
Abstract
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.
URI
http://ir.ymlib.yonsei.ac.kr/handle/22282913/140750
DOI
10.1002/jbm.a.35414
Appears in Collections:
1. 연구논문 > 2. College of Dentistry > Dept. of Oral Biology
Yonsei Authors
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Link
 http://onlinelibrary.wiley.com/doi/10.1002/jbm.a.35414/abstract
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