μct50

To avoid unnecessary animal sacrifice, data for six rats that underwent calvarial bone removal but were not otherwise treated (unfilled control Group U), and five rats that underwent insertion of a gelatin sponge following bone removal (Group G), were obtained from a previous study (GWNU-2018-14) [5 (link)]. The experimental conditions (weight, source, genetic background, age of these animals, housing and surgical procedure, etc.) of previous study (GWNU-2018-14) were identical to the current experiment (GWNU-2022-7).
Calvarial samples measuring 10 × 10 × 0.5 mm were placed in a sample holder and analyzed by μCT50 (Scanco Medical, Brüttisellen, Switzerland) in the Center for Research Facilities at Gangneung-Wonju National University. The position of each sample was confirmed. Raw images were analyzed to determine regions of interest (ROI) based on the shape and size of the original defect. The bone volume (BV) in each ROI was calculated by a CT Analyzer (version1.17.7.2+, Skyscan, Kartuizersweg, Belgium). The images were analyzed with lower and upper grayscale thresholds set to 48 and 255, respectively.

This study was approved by the Medical Ethics Committee of West China Stomatology Hospital, Sichuan University (WCHSIRB-D-2020-388). Forty-three mandibular incisors that had been extracted for reasons unrelated to this study were collected. The inclusion criteria were as follows: (1) teeth with one single canal, (2) no significant calcification or internal resorption defects, (3) no significant external root defects. Teeth with cracks, immature apices, root external defects, or coronal fillings were excluded from this study. Each tooth was scanned in a micro-CT scanner (μCT-50; Scanco Medical, Bassersdorf, Switzerland). The scanner parameters were set as 90 kV, 88μA, 8 W, resulting in an image with a 24 μm voxel size. The intracanal anatomy was confirmed according to the micro-CT scanning.

All the protocols were reviewed and approved by the Institutional Animal Care and Use Committees at Sichuan University. All experiments were carried out in accordance with the guidelines and regulations of the Institutional Animal Care and Use Committees at Sichuan University. SIRT6 knockout mice were generated as described by the Jackson Laboratory (JAX^® Mice and Services, 006050, Bar Harbor, ME, USA), and the genotyping was followed the instructions of the Jackson Laboratory. The homozygous SIRT6 mice (129/SvJ) were bred from the pairs of heterozygous SIRT6 mice. 3-week old male mice were used for this study. Bone histomorphometric analysis was conducted at State Key Laboratory of Oral Diseases, Sichuan University. For μ-CT analysis, the third lumbar vertebras and the femurs were scanned using μ-CT Scanner (μ-CT50, Scanco, Bassersdorf, Zurich, Switzerland), operated at 60 kV, 165 μA, 450 ms exposure time and 10-μm resolution. We used standardized nomenclature for the bone parameters measured according to the μ-CT Scanner protocol.

Alveolar bones with teeth dissected from mice were fixed in 4% paraformaldehyde (PFA) for 48 h. Use micro-CT (μCT50, Scanco Medical, Zurich, Switzerland) for tissue tomography and output in DICOM format. Image data were reconstructed and analyzed using the Mimics 13.0 software.

The fractions obtained from the ethanol extract were subjected to antiosteoporosis assay on OVX-induced osteoporosis in mice. Forty female C57/BL6 mice aged 8 weeks (21.2 ± 1.3 g) were obtained from the Shanghai Laboratory Animal Center (SLAC, Shanghai, China), and the experiments were approved by the Institutional Animal Care and Use Committee of Shanghai Tongji University. Animals in the OVX group were ovariectomized bilaterally, whereas the control group mice were sham-operated. One week after operation, the OVX mice were orally administered with 100 mg/kg n-hexane (FCI-H), ethyl acetate (FCI-E), or n-butanol (FCI-B) fraction daily. Subsequent to 8-week treatment, a micro-computed tomography (micro-CT) system (μCT50, Scanco Medical, Bassersdorf, Switzerland) at a resolution of 10 μm was employed for scanning the microstructural indices of the distal femurs including the bone mineral density (BMD) of the trabecular bone, bone volume/tissue volume (BV/TV), trabecular space (Tb.Sp), and trabecular number (Tb.N).

The mandibles of 3-week-old (3WK) and 8-week-old (8WK) female mice were dissected and stored in ethanol subject to µCT scanning (μCT50; Scanco Medical Ag, Bassersdorf, Switzerland), and subsequently reconstructed and analyzed with Materialise Mimics Innovation Suite Medical 21.0 software (Materialise, Leuven, Belgium).