Abstract
Bone tissue engineering and bone scaffold development represent two challenging fields in orthopaedic research. Micro-computed tomography (mCT) allows non-invasive measurement of these scaffolds’ properties in vivo. However, the lack of standardized mCT analysis protocols and, therefore, the protocols’ user-dependency make interpretation of the reported results difficult. To overcome these issues in scaffold research, we introduce the Heidelberg-mCT-Analyzer. For evaluation of our technique, we built 10 bone-inducing scaffolds, which underwent mCT acquisition before ectopic implantation (T0) in mice, and at explantation eight weeks thereafter (T1). The scaffolds’ three-dimensional reconstructions were automatically segmented using fuzzy clustering with fully automatic level-setting. The scaffold itself and its pores were then evaluated for T0 and T1. Analysing the scaffolds’ characteristic parameter set with our quantification method showed bone formation over time. We were able to demonstrate that our algorithm obtained the same results for basic scaffold parameters (e.g. scaffold volume, pore number and pore volume) as other established analysis methods. Furthermore, our algorithm was able to analyse more complex parameters, such as pore size range, tissue mineral density and scaffold surface. Our imaging and post-processing strategy enables standardized and user-independent analysis of scaffold properties, and therefore is able to improve the quantitative evaluations of scaffold-associated bone tissue-engineering projects.
Highlights
Clinical routine in orthopaedic and trauma surgery demands bone substitutes for the treatment of critical size defects, prosthesis loosening, non-union, infection of bone or defects following tumour resection [1,2,3]
In our standard tissue-engineering protocols, we perform in vivo bone formation of human mesenchymal stem cells derived from iliac crest aspirate based on ectopic implantation of hMSCcoated scaffolds under stimulation with bone morphogenetic protein-7 (BMP-7) in severe combined immunodeficiency (SCID) mice (Charles River, Wilmington, MA, USA)
In order to show potential differences in construct properties related to bone formation over time, we analysed and compared construct properties of T0 and T1 using our novel micro-computed tomography (mCT) analysis algorithm
Summary
Clinical routine in orthopaedic and trauma surgery demands bone substitutes for the treatment of critical size defects, prosthesis loosening, non-union, infection of bone or defects following tumour resection [1,2,3]. Porosity, pore volume and surface structure seem to be crucial parameters for characterizing the properties of scaffolds that increase bone healing and regeneration [4,5]. Micro-computed tomography (mCT) analysis is capable of depicting common scaffold parameters (table 1), so mCT seems to be a reliable tool for the non-invasive analysis of scaffold structure. Recent studies show that data from mCT are significantly correlated with results from histomorphometry, the current gold standard for analysis of bone and tissue structure [8]. MCT could be used as an additional tool to histomorphometry in experimental and clinical settings— for the analysis of physiological bone tissue and for the evaluation of bone scaffolds and substitutes [6,8]. The settings to achieve optimum performance of the mCT-device are well described and simplify the use of mCT in experimental designs [6]
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