title: Controlled differentiation of human bone marrow stromal cells using magnetic nanoparticle technology.
abstract: Targeting and differentiating stem cells at sites of injury and repair is an exciting and promising area for disease treatment and reparative medicine. We have investigated remote magnetic field activation of magnetic nanoparticle-tagged mechanosensitive receptors on the cell membrane of human bone marrow stromal cells (HBMSCs) for use in osteoprogenitor cell delivery systems and activation of differentiation in vitro and in vivo toward an osteochondral lineage. HBMSC-labeled with magnetic beads coated with antibodies or peptides to the transmembrane ion channel stretch activated potassium channel (TREK-1) or arginine&#x2013;glycine&#x2013;aspartic acid were cultured in monolayer or encapsulated into polysaccharide alginate/chitosan microcapsules. Upregulation in gene expression was measured in magnetic particle-labeled HBMSCs in response to TREK-1 activation over a short period (7 days) with an increase in mRNA levels of Sox9, core binding factor alpha1 (Cbfa1), and osteopontin. Magnetic particle-labeled HBMSCs encapsulated into alginate chitosan capsules were exposed to magnetic forces both in vitro and in vivo intermittently for 21 days. After 21 days the encapsulated, magnetic particle-labeled HBMSCs in vivo were viable as evidenced by extensive cell tracker green fluorescence. The mechanical stimulation of HBMSCs labeled with TREK-1 magnetic nanoparticle receptors enhanced expression of type-1 collagen in vitro with increases in proteoglycan matrix, core binding factor alpha1 (Cbfa1) and collagen synthesis, and extracellular matrix production and elevated the expression of type-1 and type-2 collagen in vivo. Additionally, the magnetically remote stimulation of HBMSCs labeled with magnetic nanoparticle arginine&#x2013;glycine&#x2013;aspartic acid considerably enhanced proteoglycan and collagen synthesis and extracellular matrix production and elevated the expression of type-1 and type-2 collagen in vivo and in vitro. Osteogenic mechanosensitive receptor manipulation by magnetic nanotechnology can induce the differentiation of osteoprogenitor cell populations toward an osteogenic lineage. These cell manipulation strategies offer tremendous therapeutic opportunities in soft and hard tissue repair.
authors.affiliation: Bone and Joint Research Group, Centre for Human Development, Stem Cells, and Regeneration, Institute of Developmental Sciences, University of Southampton, Southampton, United Kingdom.
authors.forename: Janos M
authors.initials: JM
authors.lastname: Kanczler
authors.forename: Harpul S
authors.initials: HS
authors.lastname: Sura
authors.forename: Julia
authors.initials: J
authors.lastname: Magnay
authors.forename: David
authors.initials: D
authors.lastname: Green
authors.forename: Richard O C
authors.initials: RO
authors.lastname: Oreffo
authors.forename: Jon P
authors.initials: JP
authors.lastname: Dobson
authors.forename: Alicia J
authors.initials: AJ
authors.lastname: El Haj
mesh_terms: D001854:Bone Marrow Cells / Q000166:cytology*; D002454:Cell Differentiation / Q000502:physiology*; D002470:Cell Survival / Q000502:physiology; D002478:Cells, Cultured; D006801:Humans; D007150:Immunohistochemistry; D058185:Magnetite Nanoparticles / Q000737:chemistry*; D036103:Nanotechnology / Q000379:methods; D009842:Oligopeptides / Q000737:chemistry; D024683:Potassium Channels, Tandem Pore Domain / Q000737:chemistry; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017154:Stromal Cells / Q000166:cytology*
chemical_list: D058185:Magnetite Nanoparticles; D009842:Oligopeptides; D024683:Potassium Channels, Tandem Pore Domain; C103742:potassium channel protein TREK-1; C047981:arginyl-glycyl-aspartic acid