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

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

Research output: Contribution to journalArticle

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.
LanguageEnglish
Article number12944
Number of pages12
JournalScientific Reports
Volume9
DOIs
Publication statusPublished - 10 Sep 2019

Fingerprint

Bioreactors
Stem cells
Gels
Collagen
Grafts
Vibrations (mechanical)
Strain hardening
Magnets
Natural frequencies
Atomic force microscopy
Bone
Damping
Proteins
Finite element method
Experiments

Keywords

  • mesenchymal stem cells
  • MSCs
  • nanoscale mechanical stimulation
  • osteogenesis

Cite this

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. / Design, construction and characterisation of a novel nanovibrational bioreactor and cultureware for osteogenesis. In: Scientific Reports. 2019 ; Vol. 9.
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Campsie, P, Childs, PG, Robertson, SN, Cameron, K, Hough, J, Salmeron-Sanchez, M, Tsimbouri, PM, Vichare, P, Dalby, MJ & Reid, S 2019, 'Design, construction and characterisation of a novel nanovibrational bioreactor and cultureware for osteogenesis' Scientific Reports, vol. 9, 12944 . https://doi.org/10.1038/s41598-019-49422-4

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

In: Scientific Reports, Vol. 9, 12944 , 10.09.2019.

Research output: Contribution to journalArticle

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AU - Campsie, Paul

AU - Childs, Peter G.

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AU - Hough, James

AU - Salmeron-Sanchez, Manuel

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AU - Dalby, Matthew J.

AU - Reid, Stuart

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