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Title: Noninvasive image analysis of 3D construct mineralization in a perfusion bioreactor
Authors: Porter, B.D.
Lin, A.S.P.
Peister, A.
Hutmacher, D. 
Guldberg, R.E.
Keywords: Bioreactor
Bone tissue engineering
Image analysis
Issue Date: May-2007
Citation: Porter, B.D., Lin, A.S.P., Peister, A., Hutmacher, D., Guldberg, R.E. (2007-05). Noninvasive image analysis of 3D construct mineralization in a perfusion bioreactor. Biomaterials 28 (15) : 2525-2533. ScholarBank@NUS Repository.
Abstract: Although the beneficial effects of perfusion on cell-mediated mineralization have been demonstrated in several studies, the size of the mineralized constructs produced has been limited. The ability to quantify mineralized matrix formation non-invasively within 3D constructs would benefit efforts to optimize bioreactor conditions for scaling-up constructs to clinically relevant dimensions. In this study, we report a micro-CT imaging-based technique to monitor 3D mineralization over time in a perfusion bioreactor and specifically assess mechanisms of construct mineralization by quantifying the number, size, and distribution of mineralized particle formation within constructs varying in thickness from 3 to 9 mm. As expected, mineralized matrix volume and particle number increased with construct thickness. Analyzing multiple concentric volumes inside each construct indicated that a greater proportion of the mineral volume was found within the interior of the perfused constructs. Interestingly, intermediate-sized 6 mm thick constructs were found to have the highest core mineral volume fraction and the largest mineralized particles. Two complementary mechanisms of increasing total mineral volume were observed in the 6 and 9 mm constructs: increasing particle size and increasing the number of mineralized particles, respectively. The rate of mineralized matrix formation in the perfused constructs increased from 0.69 mm3/week during the first 3 weeks of culture to 1.03 mm3/week over the final 2 weeks. In contrast, the rate of mineral deposition in the static controls was 0.01 mm3/week during the first 3 weeks of culture and 0.16 mm3/week from week 3 to week 5. The ability to monitor overall construct mineralization non-invasively coupled with quantitative analysis of mineralized particle size, number, and distribution offers a powerful tool for elucidating how mineral growth mechanisms are affected by cell type, scaffold material and architecture, or bioreactor flow conditions. © 2007 Elsevier Ltd. All rights reserved.
Source Title: Biomaterials
ISSN: 01429612
DOI: 10.1016/j.biomaterials.2007.01.013
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