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X-ray microtomography measurements of bioactive glass scaffolds in rabbit femur samples at multiple stages of bone regeneration : reduction of image artefacts and a preliminary segmentation

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Title: X-ray microtomography measurements of bioactive glass scaffolds in rabbit femur samples at multiple stages of bone regeneration : reduction of image artefacts and a preliminary segmentation
Author(s): Leino, Vesa-Matti
Contributor: University of Helsinki, Faculty of Science, Fysiikan osasto
Discipline: Fysiikka
Language: English
Acceptance year: 2020
Abstract:
A series of x-ray microtomography (micro-CT) measurements was performed on a set of rabbit femur bone samples containing artificial scaffolds of bioactive glass BAG-S53P4, implanted into an intentionally induced defect, i.e. a gap, in the femur. The scaffolds, some additionally enveloped in PLGA, were supportive structures composed of small granules of bioactive glass, intended to enhance, stimulate and guide the healing and regeneration of bone. The 34 samples were harvested from the rabbits at three different stages of healing and bone regeneration: 2 weeks, 4 weeks and 8 weeks. In addition to 27 samples that contained scaffolds of BAG-S53P4 or BAG-S53P4-PLGA, which had been implanted into the femur of a rabbit, 3 scaffolds of BAG-S53P4(-PLGA) that were not implanted and 7 control samples containing inert PMMA-implants were also included in the measurements for comparison. During the healing process the bioactive glass granules are gradually dissolved into the surrounding bodily fluids and a thin reaction layer composed of silica gel forms onto the surfaces of the granules. Subsequently an additional surface layer composed of HCA, a material that closely resembles natural hydroxyapatite, is formed onto the granules. As the healing process to regenerate the bone in the gap progresses, a complex three-dimensional network of newly formed trabecular bone grows in between the granules, attaching onto the surface layers and eventually enveloping the gradually dissolving granules entirely. Ultimately, the scaffold is intended to degrade completely, and a structure of regenerated, remodeled cortical bone is expected to be formed into the volume of the initial defect. As the thicknesses of both the surface layers of the granules and the individual trabeculae of the newly formed bone are in the micrometre range, x-ray microtomography was employed to evaluate and assess the complex three-dimensional structure, consisting of trabecular bone intertwined with granules at varying stages of dissolution. By evaluating the rate of formation of these structures at three different stages, i.e. time points, of regeneration, valuable information on the effectiveness of the bioactive glass BAG-S53P4(-PLGA) for the regeneration of defected bone can be obtained. The measurements were performed at University of Helsinki’s Laboratory of Microtomography using its Nanotom-apparatus with 80kV voltage, 150microA current and a voxel size of 15micrometres. 1000 projection images per sample were used in 37 reconstructions utilizing the FBP-algorithm. Subsequent image processing to analyze and compare the samples was conducted using ImageJ. A procedure to reduce image artefacts – due to metal parts in the samples – was developed, utilizing Gaussian filtering, as well as a preliminary image segmentation scheme, utilizing Morphological filtering, to automatically separate the bone from the granules and their surface layers.
Keyword(s): X-ray Microtomography Bioactive Glass Scaffold BAG-S53P4 BAG-S53P4-PLGA Image Artefact


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