title: Polyelectrolyte multilayer coating of 3D scaffolds enhances tissue growth and gene delivery: non-invasive and label-free assessment.
abstract: Layer-by-layer (LbL) deposition is a versatile technique which is beginning to be be explored for inductive tissue engineering applications. Here, it is demonstrated that a polyelectrolyte multilayer film system composed of glycol-chitosan (Glyc-CHI) and hyaluronic acid (HA) can be used to coat 3D micro-fabricated polymeric tissue engineering scaffolds. In order to overcome many of the limitations associated with conventional techniques for assessing cell growth and viability within 3D scaffolds, two novel, real-time, label-free techniques are introduced: impedance monitoring and optical coherence phase microscopy. Using these methods, it is shown that LbL-coated scaffolds support in vitro cell growth and viability for a period of at least two weeks at levels higher than uncoated controls. These polyelectrolyte multilayer coatings are then further adapted for non-viral gene delivery applications via incorporation of DNA carrier lipoplexes. Scaffold-based delivery of the enhanced green fluorescent protein (EGFP) marker gene from these coatings is successfully demonstrated in vitro, achieving a two-fold increase in transfection efficiency compared with control scaffolds. These results show the great potential of Glyc-CHI/HA polyelectrolyte multilayer films for a variety of gene delivery and inductive tissue engineering applications.
authors.affiliation: Department of Biomedical Engineering, McGill University, Montreal, H3A 2B4, Canada.
authors.forename: Christina
authors.initials: C
authors.lastname: Holmes
authors.forename: Jamal
authors.initials: J
authors.lastname: Daoud
authors.forename: Pierre O
authors.initials: PO
authors.lastname: Bagnaninchi
authors.forename: Maryam
authors.initials: M
authors.lastname: Tabrizian
mesh_terms: D000818:Animals; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D048271:Chitosan / Q000737:chemistry / Q000494:pharmacology*; D018014:Gene Transfer Techniques; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D006820:Hyaluronic Acid / Q000737:chemistry / Q000494:pharmacology*; D051379:Mice; D023822:Tissue Engineering / Q000295:instrumentation* / Q000379:methods; D054457:Tissue Scaffolds
chemical_list: C118638:glycol-chitosan; D049452:Green Fluorescent Proteins; D006820:Hyaluronic Acid; D048271:Chitosan
keywords: dielectric impedance spectroscopy; gene delivery; layer-by-layer deposition; phase microscopy; tissue engineering