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Updated: Sep 15, 2025

Longitudinal Micro-Computed Tomography Image Analysis for User-Defined Region of Interest in Critical-Sized Bone Defects
Published on: June 24, 2025
Longitudinal Micro-Computed Tomography Image Analysis for User-Defined Region of Interest in Critical-Sized Bone
Anthony J Yosick1, Bei Liu2, Victor Z Zhang3
1Department of Biomedical Engineering, University of Rochester; Center for Musculoskeletal Research, Department of Orthopaedics, University of Rochester Medical Center; ayosick@ur.rochester.edu.
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Micro-computed tomography (µCT) imaging analysis of bone volume is a necessary quantitative tool for investigating bone regeneration potential and outcomes within longitudinal in vivo studies. Established methods for bone segmentation utilize visualization software for whole bone µCT segmentation and alignment of complex anatomical structures. These segmentation protocols provide a robust, high-accuracy method for segmentation, alignment, and analysis but are limited in abilities of user-defined region of interest (ROI) analysis. We present a protocol expanding upon these methods to permit user-defined ROI bone volume analysis surrounding a critical-sized bone defect. The user-defined ROI surrounding the defect can be analyzed over time for in vivo longitudinal studies. Herein, we investigate µCT images of three unique rat specimens each implanted with a polycaprolactone (PCL) control scaffold. Models are analyzed by three users (2 experienced and 1 novice) at time points of 0 and 6 weeks to illustrate the ability to measure an ROI surrounding a critical-sized defect throughout a longitudinal study.

