title: Osteogenesis of mesenchymal stem cells by nanoscale mechanotransduction.
abstract: It is likely that mesenchymal stem cells will find use in many autologous regenerative therapies. However, our ability to control cell stem growth and differentiation is presently limited, and this is a major hurdle to the clinical use of these multipotent cells especially when considering the desire not to use soluble factors or complex media formulations in culture. Also, the large number of cells required to be clinically useful is currently a hurdle to using materials-based (stiffness, chemistry, nanotopography, etc.) culture substrates. Here we give a first demonstration of using nanoscale sinusoidal mechanotransductive protocols (10-14 nm displacements at 1 kHz frequency), "nanokicking", to promote osteoblastogenesis in human mesenchymal stem cell cultures. On the basis of application of the reverse piezo effect, we use interferometry to develop the optimal stem cell stimulation conditions, allowing delivery of nanoscale cues across the entire surface of the Petri dishes used. A combination of immunofluorescence, PCR, and microarray has then been used to demonstrate osteoblastogenesis, and the arrays implicate RhoA as central to osteoblastic differentiation in agreement with materials-based strategies. We validate this with pharmacological inhibition of RhoA kinase. It is easy to envisage such stimulation protocols being up-scaled to form large-scale osteoblast bioreactors as standard cell culture plates and incubators are used in the protocol.
authors: Centre for Cell Engineering, Institute for Molecular, Cell and Systems Biology, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow G12 8QQ, United Kingdom.
authors: Habib
authors: H
authors: Nikukar
authors: Stuart
authors: S
authors: Reid
authors: P Monica
authors: PM
authors: Tsimbouri
authors: Mathis O
authors: MO
authors: Riehle
authors: Adam S G
authors: AS
authors: Curtis
authors: Matthew J
authors: MJ
authors: Dalby
mesh_terms: D002454:Cell Differentiation / Q000502:physiology; D002478:Cells, Cultured; D006801:Humans; D040542:Mechanotransduction, Cellular / Q000502:physiology*; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000502:physiology*; D036103:Nanotechnology / Q000295:instrumentation; D010006:Osteoblasts / Q000166:cytology / Q000502:physiology; D010012:Osteogenesis / Q000502:physiology*; D015398:Signal Transduction; D013314:Stress, Mechanical; D014159:Transducers; D054460:rho-Associated Kinases / Q000378:metabolism
chemical_list: D054460:rho-Associated Kinases