title: Spontaneous gene transfection of human bone cells using 3D mineralized alginate-chitosan macrocapsules.
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.
authors: Oral Biosciences, Faculty of Dentistry, The University of Hong Kong, Hong Kong, Hong Kong SAR.
authors: David W
authors: DW
authors: Green
authors: Division in Anatomy and Developmental Biology, Department of Oral Biology, Oral Science Research Center, BK21 PLUS Project, Yonsei University College of Dentistry, Seoul, Korea.
authors: Eun-Jung
authors: EJ
authors: Kim
authors: Oral Biosciences, Faculty of Dentistry, The University of Hong Kong, Hong Kong, Hong Kong SAR.|Division in Anatomy and Developmental Biology, Department of Oral Biology, Oral Science Research Center, BK21 PLUS Project, Yonsei University College of Dentistry, Seoul, Korea.
authors: Han-Sung
authors: HS
authors: Jung
mesh_terms: D000464:Alginates / Q000737:chemistry*; D001672:Biocompatible Materials / Q000737:chemistry; D001861:Bone Regeneration / Q000235:genetics; D018786:Bone Substitutes / Q000737:chemistry; D002460:Cell Line; D002478:Cells, Cultured; D048271:Chitosan / Q000737:chemistry*; D017930:Genes, Reporter; D020723:Glucuronic Acid / Q000737:chemistry; D006603:Hexuronic Acids / Q000737:chemistry; D006801:Humans; D008422:Materials Testing; D045164:Mesenchymal Stem Cell Transplantation; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism*; D008903:Minerals / Q000737:chemistry; D055758:SOX9 Transcription Factor / Q000235:genetics; D014162:Transfection / Q000379:methods*
chemical_list: D000464:Alginates; D001672:Biocompatible Materials; D018786:Bone Substitutes; D006603:Hexuronic Acids; D008903:Minerals; D055758:SOX9 Transcription Factor; C527932:SOX9 protein, human; D020723:Glucuronic Acid; D048271:Chitosan
keywords: alginate; calcium phosphate; chitosan; gene transfection; nonviral