[en] In order to successfully implement tissue engineered (TE) constructs as part of a clinical therapy, it is necessary to first develop and validate quality control tools that will ensure accurate and consistent TE construct release specifications. Hence advanced methods to monitor TE construct properties need to be further developed. In this study we showed proof of concept for contrast enhanced nanofocus computed tomography (CE-nanoCT) as a 'whole-construct' imaging technique with non-invasive potential that enables 3D visualization and quantification of in vitro engineered extracellular matrix (ECM) in TE constructs. In particular we performed a 3D quantitative and qualitative structural and spatial assessment of the in vitro engineered ECM, formed during static and perfusion bioreactor cell culture in 3D TE scaffolds, using two contrast agents, namely Hexabrix(R) and phosphotungstic acid (PTA). CE-nanoCT image data were validated by comparison to Live/Dead viability/cytotoxicity and picrosirius red staining data, and to the net dry weight of the TE constructs. When using Hexabrix(R) as contrast agent, ECM volume fitted linearly with net dry ECM weight independent from the flow rate used. When using PTA as contrast agent, CE-nanoCT data showed pronounced distinction between flow conditions when compared to both net dry weight and picrosirius red staining data although linearity was maintained, indicating culture-specific structural ECM differences. This was attributed to the binding specificity of this contrast agent. This novel type of information can contribute to optimize bioreactor culture conditions and potentially critical quality characteristics of TE constructs such as ECM quantity and homogeneity, facilitating the gradual transformation of 'TE constructs' in well characterized 'TE products'.
Disciplines :
Engineering, computing & technology: Multidisciplinary, general & others
Author, co-author :
Papantoniou, Ioannis; KU Leuven > Skeletal Biology and Engineering Research Center
Sonnaert, Maarten; KU Leuven > Department of Metallurgy and Materials Engineering
Geris, Liesbet ; Université de Liège - ULiège > Département d'aérospatiale et mécanique > Génie biomécanique
Luyten, Frank Prosper; KU Leuven > Skeletal Biology and Engineering Research Center
Schrooten, Jan; KU Leuven > Department of Metallurgy and Materials Engineering
Kerckhofs, Greet ; Université de Liège - ULiège > Département d'aérospatiale et mécanique > Génie biomécanique
Language :
English
Title :
Three dimensional characterization of tissue-engineered constructs by contrast enhanced nanofocus computed tomography.
I.P. is funded by the ENDEAVOUR project G.0982.11N of the FWO;M.S. is supported by a Ph.D. grant of the Agency for Innovation by Science and Technology (IWT/111457). G.K. and L.G. acknowledge support by the European Research Council under the European Union's Seventh Framework Program (FP7/2007–2013)/ERC grant agreement n°279100.
Funders :
FWO - Fonds Wetenschappelijk Onderzoek Vlaanderen [BE] MITA - Mokslo, Inovacijų ir Technologijų Agentūra [LT] CER - Conseil Européen de la Recherche [BE]
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