title: Post-transcriptional regulation in osteoblasts using localized delivery of miR-29a inhibitor from nanofibers to enhance extracellular matrix deposition.
abstract: MicroRNAs are important post-transcriptional regulators of skeletal biology, and miRNA-based therapeutics have the potential to aid bone repair. However, efficient tools for delivering miRNA mimics or inhibitors to specific target tissues are limited. Polymeric nanofibers closely mimic natural extracellular matrix (ECM) morphology, and are attractive candidates for supporting delivery of cells and bone-anabolic reagents. It is hypothesized that gelatin nanofibers could be used for the localized transient delivery of miRNA-based therapeutics, using miR-29a inhibitor as a prototype to increase ECM deposition. miR-29 family members are negative regulators of ECM synthesis, targeting the mRNAs of selected collagens and osteonectin/SPARC. Inhibiting miR-29 activity may therefore increase ECM production by cells. miR-29a inhibitor-loaded gelatin nanofibers, prepared by electrospinning, demonstrated continuous release of miRNA inhibitor over 72h. Pre-osteoblastic murine MC3T3-E1 cell line seeded on miR-29a inhibitor-loaded nanofibers synthesized more osteonectin, indicating efficient inhibitor delivery. These cells also displayed increased Igf1 and Tgfb1 mRNA. Moreover, primary bone marrow stromal cells from transgenic pOBCol3.6cyan reporter mice, grown on miR-29a inhibitor scaffolds, displayed increased col3.6 cyan expression as well as collagen production. This study demonstrates that ECM mimicking nanostructured scaffolds, in conjunction with bioactive miRNA-based therapeutics, may serve as a novel platform for developing biologically active localized cell delivery systems.
authors.affiliation: Department of Orthopedic Surgery, University of Connecticut Health Center, 263 Farmington Avenue, Farmington, CT 06030, USA; Institute for Regenerative Engineering, University of Connecticut Health Center, 263 Farmington Avenue, Farmington, CT 06030, USA.
authors.forename: Eric N
authors.initials: EN
authors.lastname: James
authors.affiliation: Center for Molecular Medicine, University of Connecticut Health Center, 263 Farmington Avenue, Farmington, CT 06030, USA.
authors.forename: Anne M
authors.initials: AM
authors.lastname: Delany
authors.affiliation: Department of Orthopedic Surgery, University of Connecticut Health Center, 263 Farmington Avenue, Farmington, CT 06030, USA; Institute for Regenerative Engineering, University of Connecticut Health Center, 263 Farmington Avenue, Farmington, CT 06030, USA; Department of Materials Science and Engineering and Department of Biomedical Engineering, University of Connecticut Storrs, CT 06268, USA. Electronic address: nair@uchc.edu.
authors.forename: Lakshmi S
authors.initials: LS
authors.lastname: Nair
mesh_terms: D000818:Animals; D041702:BALB 3T3 Cells; D005109:Extracellular Matrix / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000502:physiology; D051379:Mice; D035683:MicroRNAs / Q000037:antagonists & inhibitors* / Q000378:metabolism; D053769:Nanocapsules / Q000008:administration & dosage / Q000737:chemistry* / Q000648:ultrastructure; D057139:Nanofibers / Q000737:chemistry* / Q000648:ultrastructure; D009841:Oligonucleotides / Q000008:administration & dosage* / Q000737:chemistry; D010006:Osteoblasts / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D012323:RNA Processing, Post-Transcriptional / Q000187:drug effects / Q000502:physiology
chemical_list: C547014:MIRN29 microRNA, mouse; D035683:MicroRNAs; D053769:Nanocapsules; D009841:Oligonucleotides
keywords: Bone; Extracellular matrix; Nanofibers; Osteonectin; microRNA