title: A novel collagen-nanohydroxyapatite microRNA-activated scaffold for tissue engineering applications capable of efficient delivery of both miR-mimics and antagomiRs to human mesenchymal stem cells.
abstract: Manipulation of gene expression through the use of microRNAs (miRNAs) offers tremendous potential for the field of tissue engineering. However, the lack of sufficient site-specific and bioactive delivery systems has severely hampered the clinical translation of miRNA-based therapies. In this study, we developed a novel non-viral bioactive delivery platform for miRNA mimics and antagomiRs to allow for a vast range of therapeutic applications. By combining nanohydroxyapatite (nHA) particles with reporter miRNAs (nanomiRs) and collagen-nanohydroxyapatite scaffolds, this work introduces the first non-viral, non-lipid platform to date, capable of efficient delivery of mature miRNA molecules to human mesenchymal stem cells (hMSCs), a particularly difficult cell type to transfect effectively, with minimal treatment-associated cytotoxicity. Firstly, miRNAs were successfully delivered to hMSCs in monolayer, with internalisation efficiencies of 17.4 and 39.6% for nanomiR-mimics and nanoantagomiRs respectively, and both nanomiR-mimics and nanoantagomiRs yielded sustained interfering activity of greater than 90% in monolayer over 7 days. When applied to 3D scaffolds, significant RNA interference of 20% for nanomiR-mimics and 88.4% for nanoantagomiRs was achieved with no cytotoxicity issues over a 7 day period. In summary, in-house synthesised non-viral nHA particles efficiently delivered reporter miRNAs both in monolayer and on scaffolds demonstrating the immense potential of this innovative miRNA-activated scaffold system for tissue engineering applications.
authors.affiliation: Tissue Engineering Research Group, Department of Anatomy, Royal College of Surgeons in Ireland, 123 St. Stephens Green, Dublin 2, Ireland; Trinity Centre for Bioengineering, Trinity College Dublin, College Green, Dublin 2, Ireland; Advanced Materials and Bioengineering Research (AMBER) Centre, RCSI & TCD, Dublin 2, Ireland.
authors.forename: Irene
authors.initials: I
authors.lastname: Mencía Castaño
authors.affiliation: Tissue Engineering Research Group, Department of Anatomy, Royal College of Surgeons in Ireland, 123 St. Stephens Green, Dublin 2, Ireland; Trinity Centre for Bioengineering, Trinity College Dublin, College Green, Dublin 2, Ireland; Advanced Materials and Bioengineering Research (AMBER) Centre, RCSI & TCD, Dublin 2, Ireland.
authors.forename: Caroline M
authors.initials: CM
authors.lastname: Curtin
authors.affiliation: Regenerative Medicine Institute, National University of Ireland, Galway, Ireland.
authors.forename: Georgina
authors.initials: G
authors.lastname: Shaw
authors.affiliation: Regenerative Medicine Institute, National University of Ireland, Galway, Ireland.
authors.forename: J Mary
authors.initials: JM
authors.lastname: Murphy
authors.affiliation: Tissue Engineering Research Group, Department of Anatomy, Royal College of Surgeons in Ireland, 123 St. Stephens Green, Dublin 2, Ireland; Trinity Centre for Bioengineering, Trinity College Dublin, College Green, Dublin 2, Ireland; Advanced Materials and Bioengineering Research (AMBER) Centre, RCSI & TCD, Dublin 2, Ireland. Electronic address: garryduffy@rcsi.ie.
authors.forename: Garry P
authors.initials: GP
authors.lastname: Duffy
authors.affiliation: Tissue Engineering Research Group, Department of Anatomy, Royal College of Surgeons in Ireland, 123 St. Stephens Green, Dublin 2, Ireland; Trinity Centre for Bioengineering, Trinity College Dublin, College Green, Dublin 2, Ireland; Advanced Materials and Bioengineering Research (AMBER) Centre, RCSI & TCD, Dublin 2, Ireland. Electronic address: fjobrien@rcsi.ie.
authors.forename: Fergal J
authors.initials: FJ
authors.lastname: O'Brien
mesh_terms: D002470:Cell Survival; D002478:Cells, Cultured; D003094:Collagen / Q000008:administration & dosage*; D017886:Durapatite / Q000008:administration & dosage*; D024601:Glyceraldehyde 3-Phosphate Dehydrogenase (NADP+) / Q000235:genetics; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000378:metabolism*; D035683:MicroRNAs / Q000008:administration & dosage*; D023822:Tissue Engineering; D054457:Tissue Scaffolds
chemical_list: D035683:MicroRNAs; C509317:nano-hydroxyapatite-collagen; D003094:Collagen; D017886:Durapatite; D024601:Glyceraldehyde 3-Phosphate Dehydrogenase (NADP+)
keywords: Collagen-based scaffolds; Human mesenchymal stem cells; MicroRNA; Nanohydroxyapatite particles; Tissue engineering