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Multifactorial Optimization of Contrast-Enhanced Nanofocus Computed Tomography for Quantitative Analysis of Neo-Tissue Formation in Tissue Engineering Constructs

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Sonnaert, Maarten Kerckhofs, Greet [UCL] Papantoniou, Ioannis Van Vlierberghe, Sandra Boterberg, Veerle Geris, Liesbet

To progress the fields of tissue engineering (TE) and regenerative medicine, development of quantitative methods for non-invasive three dimensional characterization of engineered constructs (i.e. cells/tissue combined with scaffolds) becomes essential. In this study, we have defined the most optimal staining conditions for contrast-enhanced nanofocus computed tomography for three dimensional visualization and quantitative analysis of in vitro engineered neo-tissue (i.e. extracellular matrix containing cells) in perfusion bioreactor-developed Ti6Al4V constructs. A fractional factorial ‘design of experiments’ approach was used to elucidate the influence of the staining time and concentration of two contrast agents (Hexabrix® and phosphotungstic acid) and the neo-tissue volume on the image contrast and dataset quality. Additionally, the neo-tissue shrinkage that was induced by phosphotungstic acid staining was quantified to determine the operating window within which this contrast agent can be accurately applied. For Hexabrix® the staining concentration was the main parameter influencing image contrast and dataset quality. Using phosphotungstic acid the staining concentration had a significant influence on the image contrast while both staining concentration and neo-tissue volume had an influence on the dataset quality. The use of high concentrations of phosphotungstic acid did however introduce significant shrinkage of the neo-tissue indicating that, despite sub-optimal image contrast, low concentrations of this staining agent should be used to enable quantitative analysis. To conclude, design of experiments allowed us to define the most optimal staining conditions for contrast-enhanced nanofocus computed tomography to be used as a routine screening tool of neo-tissue formation in Ti6Al4V constructs, transforming it into a robust three dimensional quality control methodology.

Document type Article de périodique (Journal article) – Article de recherche
Access type Accès libre
Publication date 2015
Language Anglais
Journal information "PLoS One" - Vol. 10, no.6, p. 1-17 (2015)
Peer reviewed yes
Publisher Public Library of Sciene
issn 1932-6203
Publication status Publié
Affiliations KUL - Division of Skeletal Tissue Engineering
KUL - Skeletal Biology and Engineering Research Center
KUL - Department of Materials Engineering
KUL - Department of Mechanical Engineering
ULg - Biomechanics Research Unit
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Bibliographic reference Sonnaert, Maarten ; Kerckhofs, Greet ; Papantoniou, Ioannis ; Van Vlierberghe, Sandra ; Boterberg, Veerle ; et. al. Multifactorial Optimization of Contrast-Enhanced Nanofocus Computed Tomography for Quantitative Analysis of Neo-Tissue Formation in Tissue Engineering Constructs. In: PLoS One, Vol. 10, no.6, p. 1-17 (2015)
Permanent URL http://hdl.handle.net/2078/194653