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Orthopaedic Proceedings
Vol. 100-B, Issue SUPP_15 | Pages 17 - 17
1 Nov 2018
Dalgarno K Benning M Partridge S Tulah A Ahmed S Dickinson A Genever P Pearson R Feichtinger G Loughlin J Ferreira-Duarte A
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This paper reports on a proof of concept project funded by the UK National Council for the Replacement, Refinement and Reduction of Animals in Research (NC3Rs), with the aim of developing an in vitro model to recapitulate the human osteoarthritic joint, based on a multiple human cell type co-culture system, for research and drug development in OA. The targets were: (i) the development of a cell culture platform that could produce a mixed stable cell culture of cell types that represent the key components of the human joint: synoviocytes – type I and type II; osteoblasts; osteoclasts; chondrocytes/cartilage or cartilage-like matrix; adipocytes; and immune cells. (ii) demonstration of cell phenotype stability and viability for at least 72 hours. In order to establish the cell culture platform we have developed an eight-channel cell printer, capable of accurately and reliably printing the required cell types to create osteochondral and synovial cell types within a transwell system. Two different sets of cells have been developed and processed using the cell printer: a set based on using an immortalised hTERT MSC line to create osteoblasts, chondrocytes and adipocytes, with commercial cells lines providing the other cell types, and a set obtained from tissue excised during orthopaedic surgery. This gives both a repeatable set of cells with which to undertake mode of action studies, and a bank of cell sets which will be representative of different stages of osteoarthritis. The co-cultures have been immunohistochemically assessed in order to demonstrate maintenance of phenotype.


Orthopaedic Proceedings
Vol. 100-B, Issue SUPP_14 | Pages 115 - 115
1 Nov 2018
Müller S Nicholson L Jone E Dickinson A Dalgarno K Wang X
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Mesenchymal stromal cells (MSCs) are widely used in clinical trials for the treatment of many bone defects. Apatite-wollastonite glass ceramic (A-W) is an osteoconductive biomaterial shown to be compatible with MSCs. This is the first study comparing the osteogenic potential of two MSC populations, heterogeneous plastic adherence MSCs (PA-MSCs) and CD271-enriched MSCs (CD271-MSCs), when cultured on A-W 3D scaffold. The paired MSC populations were assessed for their attachment, growth kinetics and ALP activity using confocal or scanning electron microscopy and the quantifications of DNA contents and p-nitrophenyl (pNP) production. While the PA-MSCs and CD271-MSCs had similar expansion and tri-lineage differentiation capacity during standard 2D culture, they showed different proliferation kinetics when seeded on the A-W scaffolds. PA-MSCs displayed a well-spread attachment with more elongated morphology compared to CD271-MSCs, signifying a different level of interaction between the cell populations and the scaffold surface. PA-MSCs also fully integrated into the scaffold surface and showed a stronger propensity for osteogenic differentiation on the A-W scaffold as indicated by higher ALP activity than CD271-MSCs. Furthermore, A-W scaffold seeded uncultured bone marrow mononuclear cells (BM-MNCs) demonstrated a higher proliferation rate and greater ALP activity compared to freshly isolated CD271-enriched BM-MNCs. Our findings suggest that enrichment of CD271-positive population is not beneficial for osteogenesis when the cells are seeded on A-W scaffold. Furthermore, unselected heterogeneous MSCs or BM-MNCs are more promising for A-W scaffold-based bone regeneration, providing novel insight with potential clinical implications in regenerative medicine for bone defects using an innovative tissue engineering approach.


Orthopaedic Proceedings
Vol. 99-B, Issue SUPP_2 | Pages 76 - 76
1 Jan 2017
Marter A Pierron F Dickinson A Browne M
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Polymer foams have been used extensively in the testing and development of orthopaedic devices and for verification of computational models. Their use is often preferred over cadaver and animal models due to being relatively inexpensive and their consistent material properties. Successful validation of such models requires accurate material/mechanical data. The assumed range of compressive moduli, provided in the sawbones technical sheet, is 16 MPa to 1.15 GPa depending on the density of foam. In this investigation, we apply two non-contact measurement techniques (digital volume correlation (DVC) and optical surface extensometry) to assess the validity of these reported values. It is thought that such non-contact methods remove mechanical extensometer errors (slippage, misalignment) and restrict the effect of test-machine end-artifacts (friction, non-uniform loading, platen flexibility). This is because measurement is taken directly from the sample, and hence material property assessment should be more accurate. Use of DVC is advantageous as full field strain measurement is possible, however test time and cost is significantly higher than extensometry. Hence, the study also sought to assess the viability of optical extensometry for characterising porous materials.

Testing was conducted on five 20 mm cubic samples of 0.32g/cc (20 pcf) solid rigid polyurethane foam (SAWBONESTM). The strain behaviour was characterised by incremental loading via an in situ loading rig. Loading was performed in 0.1 mm increments for 8 load steps with scans between loading steps. Full field strain measurement was performed on one sample by micro focus tomography (muvis centre, Southampton) and subsequent DVC (DaVis, Lavision). Calculation of Young's modulus and Poisson's ratio was then preformed through use of the virtual fields method. These results were subsequently corroborated by use of optical extensometry (MatchID). To account for heterogeneities, axial strain measurements were averaged from six points on the front and rear surfaces. A computationally derived correction factor was then applied to account for through volume strain variations. In each test compressive displacement was applied to 900N (∼2MPa) to remain within the linear elastic region.

Significant variability of individual strain measurements were observed from extensometry measurements on the same sample, indicating non-uniform loading did occur in all samples. However by averaging across multiple points linear loading profiles were identified. For all non-contact methods the calculated elastic moduli were found to range between 331–428 MPa whilst the approximated modulus based on cross head displacement was ∼210 MPa. The optical-extensometry gave a considerably higher modulus (p = 0.047) than the DVC results as only surface measurements were made. However, following computational based correction values converged within 6% of one another. Both the DVC and point-tracking results (p = 0.001) indicated substantially higher compressive modulus (137%) than the manufacturer provided properties.

This study demonstrates that methods of measuring displacement data on of cellular foams must be carefully considered, as artefacts can lead to significant errors of up to 137%, and such errors may falsely influence the design and validation of tested devices.