The extracted skulls were studied using a μCT machine (SKYSCAN1173, Ver. 1.6, BRUKER-CT, KONTICH, Belgium) for imaging. The pre-set imaging conditions were 60 μA tube current, 130 kVp tube voltage, 500 ms exposure time, 1 mm aluminum filter, and 0.3° rotation angle. The pixel size was 13.85 μm, and the number of pixels was 2240 × 2240. With the help of a μCT scan, a total of 800 raw high-resolution images were obtained. The NRecon program (Ver 1.7.0.5, BRUKER-CT, KONTICH, Belgium) was used for the cross-sectional reconfiguration. The Dataviewer program (Ver. 1.5.1.3, BRUKER-CT, KONTICH, Belgium) and the Ct-VOX program (Ver. 1.14.4.2, BRUKER-CT, KONTICH, Belgium) were used for 3D reconstruction. The volume of newly formed bone inside the scaffolds was calculated using the difference in grayscale level. In the program, since the HU (Housefield unit) of the scaffold was 400, the HU of the cortical bone was 600 or more, and that of the soft tissue was 100 or less, the grayscale threshold of the new bone was set to 150–350 HU. Newly formed bone volume in the scaffolds was calculated using these programs, according to the following calculation:
Ct vox program
Manufactured by Bruker
Sourced in Belgium
Ct-VOX is a software program developed by Bruker for the analysis and visualization of computed tomography (CT) data. The core function of Ct-VOX is to provide users with tools for the reconstruction, segmentation, and three-dimensional (3D) visualization of CT scan data.
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3 protocols using ct vox program
1
Micro-CT Analysis of Bone Scaffold
2
Micro-CT Analysis of Murine Tumor Legs
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Hind legs with tumors were isolated from the sacrificed mice after photothermal treatment. The hind legs of mice were placed in a scanning rack and analyzed using the Siemens Biograph 3D micro-CT device (Skyscan 1076, Antwerp, Belgium). After scanning, the 3D model was reconstructed and evaluated using the CTVox program (Bruker micro-CT NV, Antwerp, Belgium).
3
Osteoporosis Treatment with ASC-EVs
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Eight‐week‐old female ICR (CD‐1) mice were purchased from Japan SLC, Inc., and 24 mice were bilaterally OVX while six mice underwent the sham operation. All animal experiments were performed with approval from the Institutional Review Board of Hanyang University. Three weeks after OVX, mice were randomly divided into four groups (n = 6) and intravenously injected with ASC‐EVs (1 × 108 or 5 × 108 particles/100 μl PBS, thrice a week for 2 weeks), hASCs (5 × 105 cells/100 μl PBS, once a week for 2 weeks) (Cho et al., 2012 (link)), or an equal volume of PBS, respectively. At 5 weeks after injection, mice were sacrificed, and the left femurs of each group were harvested. The specimens were fixed in 4% paraformaldehyde for 2 days and subsequently scanned using micro‐computed tomography (μCT; Bruker, Kontich, Belgium) at 6.75 μm resolution. Three‐dimensional (3D) images were generated using the CTvox program (Bruker, Billerica, MA, USA).
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