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    Design, construction and characterisation of a novel nanovibrational bioreactor and cultureware for osteogenesis

    Year: 2019

    Journal: Sci Rep, Volume 9, SEP 10

    Authors: Campsie, Paul; Childs, Peter G.; Robertson, Shaun N.; Cameron, Kenny; Hough, James; Salmeron-Sanchez, Manuel; Tsimbouri, Penelope M.; Vichare, Parag; Dalby, Matthew J.; Reid, Stuart

    Organizations: STFCUK Research & Innovation (UKRI)Science & Technology Facilities Council (STFC) [ST/N005406/2, ST/L502509/1]; BBSRCUK Research & Innovation (UKRI)Biotechnology and Biological Sciences Research Council (BBSRC) [BB/N012690/1, BB/P00220X/1]; EPSRCUK Research & Innovation (UKRI)Engineering & Physical Sciences Research Council (EPSRC) [EP/N013905/1, EP/P001114/1]; Find A Better Way, SUPA; Royal SocietyRoyal Society of LondonEuropean Commission; Royal Society of Edinburgh (RSE); NHS Greater Glasgow Clyde; Linn Products Ltd; University of the West of Scotland; University of Glasgow; University of Strathclyde

    In regenerative medicine, techniques which control stem cell lineage commitment are a rapidly expanding field of interest. Recently, nanoscale mechanical stimulation of mesenchymal stem cells (MSCs) has been shown to activate mechanotransduction pathways stimulating osteogenesis in 2D and 3D culture. This has the potential to revolutionise bone graft procedures by creating cellular graft material from autologous or allogeneic sources of MSCs without using chemical induction. With the increased interest in mechanical stimulation of cells and huge potential for clinical use, it is apparent that researchers and clinicians require a scalable bioreactor system that provides consistently reproducible results with a simple turnkey approach. A novel bioreactor system is presented that consists of: a bioreactor vibration plate, calibrated and optimised for nanometre vibrations at 1 kHz, a power supply unit, which supplies a 1 kHz sine wave signal necessary to generate approximately 30 nm of vibration amplitude, and custom 6-well cultureware with toroidal shaped magnets incorporated in the base of each well for conformal attachment to the bioreactor's magnetic vibration plate. The cultureware and vibration plate were designed using finite element analysis to determine the modal and harmonic responses, and validated by interferometric measurement. This helps ensure that the vibration plate and cultureware, and thus collagen and MSCs, all move as a rigid body, avoiding large deformations close to the resonant frequency of the vibration plate and vibration damping beyond the resonance. Assessment of osteogenic protein expression was performed to confirm differentiation of MSCs after initial biological experiments with the system, as well as atomic force microscopy of the 3D gel constructs during vibrational stimulation to verify that strain hardening of the gel did not occur. This shows that cell differentiation was the result of the nanovibrational stimulation provided by the bioreactor alone, and that other cell differentiating factors, such as stiffening of the collagen gel, did not contribute.
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