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Avoiding artefacts in MicroCT imaging of collagen scaffolds: Effect of phosphotungstic acid (PTA)-staining and crosslink density

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posted on 2024-10-01, 09:33 authored by KA Kwon, DV Bax, JH Shepherd, RE Cameron, SM Best
X-ray micro-computed tomography (μ-CT) can be used to provide both qualitative and quantitative information on the structure of three-dimensional (3D) bioactive scaffolds. When performed in a dry state, μ-CT accurately reflects the structure of collagen-based scaffolds, but imaging in a wet state offers challenges with radiolucency. Here we have used phosphotungstic acid (PTA) as a contrast agent to visualise fully hydrated collagen scaffolds in a physiologically relevant environment. A systematic investigation was performed to understand the effects of PTA on the results of μ-CT imaging by varying sample processing variables such as crosslinking density, hydration medium and staining duration. Immersing samples in 0.3% PTA solution overnight completely stained the samples and the treatment provided a successful route for μ-CT analysis of crosslinked samples. However, significant structural artefacts were observed for samples which were either non-crosslinked or had low levels of crosslinking, which had a heterogeneous interior architecture with collapsed pores at the scaffold periphery. This work highlights the importance of optimising the choice of processing and staining conditions to ensure accurate visualisation for hydrated 3D collagen scaffolds in an aqueous medium.

Funding

Development of a Comprehensive Toolkit for Optimised Tissue Regeneration: Scaffold DOCTR

Engineering and Physical Sciences Research Council

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Citation

K.A. Kwon, D.v. Bax, J.H. Shepherd, R.E. Cameron, S.M. Best Avoiding artefacts in MicroCT imaging of collagen scaffolds: effect of phosphotungstic acid (PTA)-staining and crosslink density Bioact. Mater., 8 (2022), pp. 210-219, 10.1016/J.BIOACTMAT.2021.06.012

Author affiliation

School of Engineering, University of Leicester

Version

  • VoR (Version of Record)

Published in

Bioactive Materials

Volume

8

Pagination

210 - 219

Publisher

Elsevier BV

issn

2452-199X

eissn

2452-199X

Acceptance date

2021-06-09

Copyright date

2021

Available date

2024-10-01

Notes

Under a Creative Commons license

Spatial coverage

China

Language

eng

Data Access Statement

The underlying data for this paper may be found on the University of Cambridge Repository at https://doi.org/10.17863/CAM.70872

Rights Retention Statement

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