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

Campsie, P., Childs, P. G., Robertson, S. N., Cameron, K., Hough, J. , Salmeron-Sanchez, M. , Tsimbouri, P. M., Vichare, P., Dalby, M. J. and Reid, S. (2019) Design, construction and characterisation of a novel nanovibrational bioreactor and cultureware for osteogenesis. Scientific Reports, 9(1), 12944. (doi: 10.1038/s41598-019-49422-4) (PMID:31506561) (PMCID:PMC6736847)

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Abstract

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.

Item Type:Articles
Additional Information:Funding and fnancial 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.
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Salmeron-Sanchez, Professor Manuel and Dalby, Professor Matthew and Hough, Professor James and Childs, Dr Peter
Authors: Campsie, P., Childs, P. G., Robertson, S. N., Cameron, K., Hough, J., Salmeron-Sanchez, M., Tsimbouri, P. M., Vichare, P., Dalby, M. J., and Reid, S.
College/School:College of Medical Veterinary and Life Sciences > Institute of Molecular Cell and Systems Biology
College of Science and Engineering > School of Engineering > Biomedical Engineering
Journal Name:Scientific Reports
Publisher:Nature Publishing Group
ISSN:2045-2322
ISSN (Online):2045-2322
Copyright Holders:Copyright © 2019 The Authors
First Published:First published in Scientific Reports 9(1):12944
Publisher Policy:Reproduced under a Creative Commons License

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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
172535Development of NanoKick BioreactorMatthew DalbyBiotechnology and Biological Sciences Research Council (BBSRC)BB/N012690/1Institute of Molecular, Cell & Systems Biology
173494Rapid Bone Graft Synthesis Through Dual Piezoelectric/Nanomechaniocal StimulationMatthew DalbyBiotechnology and Biological Sciences Research Council (BBSRC)BB/P00220X/1Institute of Molecular, Cell & Systems Biology
172525Developing the NanoKick bioreactor to enable tissue engineered bone graft and use of metabolomics to identify bone specific drug candidatesMatthew DalbyEngineering and Physical Sciences Research Council (EPSRC)EP/N013905/1Institute of Molecular, Cell & Systems Biology
173192Engineering growth factor microenvironments- a new therapeutic paradigm for regenerative medicineManuel Salmeron-SanchezEngineering and Physical Sciences Research Council (EPSRC)EP/P001114/1ENG - Biomedical Engineering