AnalyzeDirect Joy E. Gerasco et al. Commensal Microbiota Effects on Craniofacial Skeletal Growth and Morphology Research Square.
Publication Date
May 31, 2023
How Analyze was Used
“Skull craniometric measurements were performed using Analyze 14.0 Bone Microarchitecture Analysis software (AnalyzeDirect, Overland Park, KS, USA). Linear measurements were carried out on reoriented 3D μCT reconstructions, with a fixed threshold of 2500 Hounsfield units (HU). For frontal and parietal bones, linear measurements were taken bilaterally and averaged. Linear measurements were performed three separate times for each outcome of interest, and the arithmetic mean of these measurements is reported for each biological replicate. Skull length (nasale to paro) is reported as a direct measurement. All other linear measurements were normalized to skull length (nasale to paro) to account for skull size differences, as reported previously. Landmarks for craniometric analyses are depicted via schematics: skull length, skull width, and skull height; skull length, nasal bone length, cranial vault bone lengths, and cranial base bone lengths. Studies were performed and are reported in accordance with guidelines for assessment of bone microstructure in rodents using μCT.
Skull BMD measurements were acquired using Analyze 14.0 Bone Microarchitecture Analysis software (AnalyzeDirect). BMD analyses were carried out using reoriented 3D μCT reconstructions. A fixed threshold of 2000 HU was used to discern mineralized tissue. BMD analysis was performed in the following volumes of interest, adapted from methods described previously[21, 46, 47]: Basioccipital bone analysis: 900 μm3 volume of interest (VOI) positioned 180 μm posterior to the junction of the basisphenoid and basioccipital bones, the spheno-occipital synchondrosis (SOS). The VOI was centered mediolaterally across the median sagittal plane. Basisphenoid bone analysis: 900 μm3 VOI positioned 360 μm anterior to the SOS and 360 μm superior from the inferior extent of the pterygoid bone/process at the intersection of the SOS. Presphenoid bone analysis: 900 μm3 VOI positioned 270 μm anterior to the junction of the basisphenoid and presphenoid bones, the intersphenoid synchondrosis (ISS). The VOI was oriented in the superior–inferior and mediolateral directions centered on the presphenoid bone. Frontal bone analysis: 1440 μm3 VOI was analyzed at the left frontal bone. The VOI was positioned 720 μm lateral to the sagittal suture and 1440 μm anterior to the coronal suture. Parietal bone analysis: 1440 μm3 VOI was analyzed at the left parietal bone. The VOI was positioned 1440 μm lateral to the sagittal suture and centered between the lambdoid and coronal sutures in the anterior–posterior plane. Interparietal bone analysis: 900 μm3 VOI positioned 900 μm anterior to the junction of the interparietal and occipital bones. The VOI was centered mediolaterally across the median sagittal plane. Landmarks and VOIs for BMD analysis are depicted via schematics: cranial vault bones (Fig. S2A) and cranial base bones (Fig. S2E). Studies were performed and are reported in accordance with guidelines for assessment of bone microstructure in rodents using μCT.”
Keywords
Bone–Microbiota Interactors, Craniometry, Preclinical Studies
Author Affiliation(s)
Medical University of South Carolina, Charleston, South Carolina, USA
