Deep behavioral phenotyping tracks functional recovery following tibia fracture in mice

IntroductionAn estimated 178 million fractures occur worldwide annually, with lower limb fractures showing high rates of poor healing, often resulting in reduced mobility and chronic pain. Bone healing and the ability to bear weight are closely tied to the mechanical stability of the fracture site....

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Main Authors: Jonathan E. Layne, Dustin M. Snapper, Molly E. Czachor, Charles Lam, Jacob D. Matityahu, Dane R. G. Lind, Matthieu Huard, Johnny Huard, Kazuhito Morioka, Julian C. Motzkin, Allan I. Basbaum, Jarret A. P. Weinrich, Chelsea S. Bahney
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Language:English
Published: Frontiers Media S.A. 2025-08-01
Series:Frontiers in Physiology
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Online Access:https://www.frontiersin.org/articles/10.3389/fphys.2025.1630155/full
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author Jonathan E. Layne
Dustin M. Snapper
Dustin M. Snapper
Molly E. Czachor
Charles Lam
Jacob D. Matityahu
Dane R. G. Lind
Matthieu Huard
Johnny Huard
Kazuhito Morioka
Julian C. Motzkin
Julian C. Motzkin
Allan I. Basbaum
Jarret A. P. Weinrich
Jarret A. P. Weinrich
Chelsea S. Bahney
Chelsea S. Bahney
author_facet Jonathan E. Layne
Dustin M. Snapper
Dustin M. Snapper
Molly E. Czachor
Charles Lam
Jacob D. Matityahu
Dane R. G. Lind
Matthieu Huard
Johnny Huard
Kazuhito Morioka
Julian C. Motzkin
Julian C. Motzkin
Allan I. Basbaum
Jarret A. P. Weinrich
Jarret A. P. Weinrich
Chelsea S. Bahney
Chelsea S. Bahney
author_sort Jonathan E. Layne
collection DOAJ
description IntroductionAn estimated 178 million fractures occur worldwide annually, with lower limb fractures showing high rates of poor healing, often resulting in reduced mobility and chronic pain. Bone healing and the ability to bear weight are closely tied to the mechanical stability of the fracture site. Although fracture stabilization is a well-established factor modulating bone repair, there remains a notable gap in sophisticated non-destructive technologies that can rapidly and objectively quantify functional recovery in preclinical settings. We introduce a novel behavioral phenotyping approach enabling rapid quantification of post-fracture weightbearing and kinematic metrics in freely behaving mice. Our goals were to identify and characterize metrics most indicative of fracture-induced behavioral impairment and to use these metrics to quantify how functional recovery is altered in mice with pin stabilized versus non-stabilized fractures. We also explore sex-specific contributions to recovery.MethodsMale and female C57BL6/J mice received mid-shaft tibial fractures that were either unstabilized or fixed with intramedullary pins; non-fractured mice served as controls. Behavioral recordings were acquired pre-fracture and throughout healing (5–35 days post-fracture). To track mice and analyze changes in paw pressure and kinematics, we performed machine learning-enabled behavioral phenotyping.ResultsOverall, mice with pin-stabilized fractures exhibited less behavioral impairment than mice with unstabilized fractures. Pin stabilization allowed increased weightbearing and produced smaller changes in kinematic metrics. By contrast, we observed only minor sex-specific differences in impairment and recovery following fracture. Our analysis revealed that functional recovery is more complex than individual parameters viewed in isolation, with different parameters identifying distinct recovery timeframes. Therefore, we developed a comprehensive, unified graph theoretic metric encompassing all behavioral parameters. This unified approach confirmed increased severity in unstabilized fractures and identified clear functional recovery windows for both fracture groups.DiscussionThis methodology forms a foundation for future mechanistic experiments focused on biological and mechanical variables influencing functional healing and enables more rapid testing of strategies to accelerate bone healing.
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spelling doaj-art-40e5e3d9d6bb485e94b5ee6ae1b263a42025-08-26T05:27:48ZengFrontiers Media S.A.Frontiers in Physiology1664-042X2025-08-011610.3389/fphys.2025.16301551630155Deep behavioral phenotyping tracks functional recovery following tibia fracture in miceJonathan E. Layne0Dustin M. Snapper1Dustin M. Snapper2Molly E. Czachor3Charles Lam4Jacob D. Matityahu5Dane R. G. Lind6Matthieu Huard7Johnny Huard8Kazuhito Morioka9Julian C. Motzkin10Julian C. Motzkin11Allan I. Basbaum12Jarret A. P. Weinrich13Jarret A. P. Weinrich14Chelsea S. Bahney15Chelsea S. Bahney16Department of Orthopaedic Surgery, UCSF Orthopaedic Trauma Institute, San Francisco, CA, United StatesDepartment of Orthopaedic Surgery, UCSF Orthopaedic Trauma Institute, San Francisco, CA, United StatesCenter for Regenerative Medicine, Steadman Philippon Research Institute, Vail, CO, United StatesCenter for Regenerative Medicine, Steadman Philippon Research Institute, Vail, CO, United StatesDepartment of Orthopaedic Surgery, UCSF Orthopaedic Trauma Institute, San Francisco, CA, United StatesDepartment of Orthopaedic Surgery, UCSF Orthopaedic Trauma Institute, San Francisco, CA, United StatesCenter for Regenerative Medicine, Steadman Philippon Research Institute, Vail, CO, United StatesCenter for Regenerative Medicine, Steadman Philippon Research Institute, Vail, CO, United StatesCenter for Regenerative Medicine, Steadman Philippon Research Institute, Vail, CO, United StatesDepartment of Orthopaedic Surgery, UCSF Orthopaedic Trauma Institute, San Francisco, CA, United StatesUCSF Department of Neurology and Weill Institute for Neuroscience, San Francisco, CA, United StatesUCSF Department of Anesthesia and Perioperative Care, San Francisco, CA, United StatesUCSF Department of Anatomy, San Francisco, CA, United StatesUCSF Department of Anesthesia and Perioperative Care, San Francisco, CA, United StatesUCSF Department of Anatomy, San Francisco, CA, United StatesDepartment of Orthopaedic Surgery, UCSF Orthopaedic Trauma Institute, San Francisco, CA, United StatesCenter for Regenerative Medicine, Steadman Philippon Research Institute, Vail, CO, United StatesIntroductionAn estimated 178 million fractures occur worldwide annually, with lower limb fractures showing high rates of poor healing, often resulting in reduced mobility and chronic pain. Bone healing and the ability to bear weight are closely tied to the mechanical stability of the fracture site. Although fracture stabilization is a well-established factor modulating bone repair, there remains a notable gap in sophisticated non-destructive technologies that can rapidly and objectively quantify functional recovery in preclinical settings. We introduce a novel behavioral phenotyping approach enabling rapid quantification of post-fracture weightbearing and kinematic metrics in freely behaving mice. Our goals were to identify and characterize metrics most indicative of fracture-induced behavioral impairment and to use these metrics to quantify how functional recovery is altered in mice with pin stabilized versus non-stabilized fractures. We also explore sex-specific contributions to recovery.MethodsMale and female C57BL6/J mice received mid-shaft tibial fractures that were either unstabilized or fixed with intramedullary pins; non-fractured mice served as controls. Behavioral recordings were acquired pre-fracture and throughout healing (5–35 days post-fracture). To track mice and analyze changes in paw pressure and kinematics, we performed machine learning-enabled behavioral phenotyping.ResultsOverall, mice with pin-stabilized fractures exhibited less behavioral impairment than mice with unstabilized fractures. Pin stabilization allowed increased weightbearing and produced smaller changes in kinematic metrics. By contrast, we observed only minor sex-specific differences in impairment and recovery following fracture. Our analysis revealed that functional recovery is more complex than individual parameters viewed in isolation, with different parameters identifying distinct recovery timeframes. Therefore, we developed a comprehensive, unified graph theoretic metric encompassing all behavioral parameters. This unified approach confirmed increased severity in unstabilized fractures and identified clear functional recovery windows for both fracture groups.DiscussionThis methodology forms a foundation for future mechanistic experiments focused on biological and mechanical variables influencing functional healing and enables more rapid testing of strategies to accelerate bone healing.https://www.frontiersin.org/articles/10.3389/fphys.2025.1630155/fullbehavioral phenotypingfracture repairfracture-related painmachine-learning image analysissexual dimorphism
spellingShingle Jonathan E. Layne
Dustin M. Snapper
Dustin M. Snapper
Molly E. Czachor
Charles Lam
Jacob D. Matityahu
Dane R. G. Lind
Matthieu Huard
Johnny Huard
Kazuhito Morioka
Julian C. Motzkin
Julian C. Motzkin
Allan I. Basbaum
Jarret A. P. Weinrich
Jarret A. P. Weinrich
Chelsea S. Bahney
Chelsea S. Bahney
Deep behavioral phenotyping tracks functional recovery following tibia fracture in mice
Frontiers in Physiology
behavioral phenotyping
fracture repair
fracture-related pain
machine-learning image analysis
sexual dimorphism
title Deep behavioral phenotyping tracks functional recovery following tibia fracture in mice
title_full Deep behavioral phenotyping tracks functional recovery following tibia fracture in mice
title_fullStr Deep behavioral phenotyping tracks functional recovery following tibia fracture in mice
title_full_unstemmed Deep behavioral phenotyping tracks functional recovery following tibia fracture in mice
title_short Deep behavioral phenotyping tracks functional recovery following tibia fracture in mice
title_sort deep behavioral phenotyping tracks functional recovery following tibia fracture in mice
topic behavioral phenotyping
fracture repair
fracture-related pain
machine-learning image analysis
sexual dimorphism
url https://www.frontiersin.org/articles/10.3389/fphys.2025.1630155/full
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