title: Manipulation of miRNA activity accelerates osteogenic differentiation of hMSCs in engineered 3D scaffolds.
abstract: Cell-based tissue engineering strategies have shown tremendous promise for the repair of bone mass deficiencies, but the efficient and appropriate induction of stem cells down osteogenic pathways remains a significant roadblock to the effective implementation of cell-based therapies. When grown in culture, human Mesenchymal Stromal/Stem Cells (hMSCs) remain multipotent, requiring specific exogenous signals to induce osteogenic differentiation. hMSCs used in transplantations, therefore, must be presented with local signals, often provided by the host's own tissues, to be directed down bone-related lineages. This process is relatively inefficient and remains difficult to control. In an effort to enhance osteogenesis, hMSCs were transfected with specific miRNA mimics and inhibitors that had originally identified for their ability to increase Alkaline Phosphatase (ALP) activity. Transfection with miRNA reagents had the effect of sensitizing hMSCs to soluble osteogenic factors, resulting in a rapid and robust induction of bone-related markers, including ALP activity and calcium deposition. Synthetic 3D tissue constructs prepared with miRNA-transfected hMSCs demonstrated similar responses to soluble osteogenic signals, suggesting that controlling miRNA activity in hMSCs can be an effective tool for enhancing the induction of osteogenesis for tissue engineering purposes.
authors.affiliation: Howard Hughes Medical Institute, University of Colorado at Boulder, CO, USA.
authors.forename: Peter D
authors.initials: PD
authors.lastname: Mariner
authors.forename: Erika
authors.initials: E
authors.lastname: Johannesen
authors.forename: Kristi S
authors.initials: KS
authors.lastname: Anseth
mesh_terms: D000469:Alkaline Phosphatase / Q000378:metabolism; D001205:Ascorbic Acid / Q000031:analogs & derivatives / Q000494:pharmacology; D015415:Biomarkers / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects / Q000235:genetics*; D002478:Cells, Cultured; D003094:Collagen / Q000378:metabolism; D003470:Culture Media / Q000494:pharmacology; D003907:Dexamethasone / Q000494:pharmacology; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000201:enzymology / Q000378:metabolism*; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D010012:Osteogenesis / Q000187:drug effects / Q000235:genetics*; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds / Q000737:chemistry*; D014162:Transfection
chemical_list: D015415:Biomarkers; D003470:Culture Media; C550570:MIRN148 microRNA, human; C552418:MIRN489 microRNA, human; D035683:MicroRNAs; C011669:ascorbate-2-phosphate; D003907:Dexamethasone; D003094:Collagen; D000469:Alkaline Phosphatase; D001205:Ascorbic Acid; D002118:Calcium