TY - JOUR
T1 - Design, construction and characterisation of a novel nanovibrational bioreactor and cultureware for osteogenesis
AU - Campsie, Paul
AU - Childs, Peter G.
AU - Robertson, Shaun N.
AU - Cameron, Kenny
AU - Hough, James
AU - Salmeron-Sanchez, Manuel
AU - Tsimbouri, Penelope M.
AU - Vichare, Parag
AU - Dalby, Matthew J.
AU - Reid, Stuart
N1 - Funding Information:
The authors would like to thank: Iain Martin, Habib Nikukar, Keith Robertson, Gilad Tiefenbrun and Ivor Tiefenbrun for their advice, Gerry O’Hare and Ross Simpson at the University of the West of Scotland for their help etching and assembling the power supply PCBs, Jim Orr and his workshop team at the University of the West of Scotland for machining the bioreactor parts and for helpful discussions on their design, CGP Engineering Limited for manufacturing the mould and carrying out the initial production run of cultureware, Milacron for advice and useful discussion on mould design and materials, and the GEO and LIGO Scientific Collaboration for their interest. Funding and financial support from STFC (ST/N005406/2, ST/L502509/1), BBSRC (BB/N012690/1, BB/P00220X/1), EPSRC (EP/N013905/1, EP/P001114/1), Find A Better Way, SUPA, the Royal Society (RS), the Royal Society of Edinburgh (RSE), NHS Greater Glasgow & Clyde, Linn Products Ltd, the University of the West of Scotland, University of Glasgow and University of Strathclyde are gratefully acknowledged.
Publisher Copyright:
© 2019, The Author(s).
PY - 2019/12/1
Y1 - 2019/12/1
N2 - 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.
AB - 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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U2 - 10.1038/s41598-019-49422-4
DO - 10.1038/s41598-019-49422-4
M3 - Article
C2 - 31506561
AN - SCOPUS:85072020346
VL - 9
JO - Scientific Reports
JF - Scientific Reports
SN - 2045-2322
IS - 1
M1 - 12944
ER -