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Related Experiment Video

Updated: Jan 7, 2026

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Critical-Size Muscle Defect Regeneration Using an Injectable Cell-Laden Nanofibrous Matrix: An Ex Vivo Mouse Hindlimb

Diego Jacho1, James Huynh1, Emily Crowe2

  • 1Department of Bioengineering, College of Engineering, University of Toledo, Toledo, OH 43606, USA.

International Journal of Molecular Sciences
|December 30, 2025
PubMed
Summary

This study presents an ex vivo mouse model for testing muscle regeneration therapies. Combining mechanical stimulation with a novel scaffold significantly enhanced muscle repair and reduced inflammation.

Keywords:
3D tissue culturinganimal welfarecollagenculturedefectengineeringex vivomechanical loadingmechanotherapymuscleorganregenerationtibialistissue

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Area of Science:

  • Biomedical Engineering
  • Regenerative Medicine
  • Tissue Engineering

Background:

  • Volumetric muscle loss injuries present significant treatment challenges due to limited regeneration.
  • Existing experimental models lack physiological relevance for studying muscle repair.
  • Developing effective regenerative strategies requires advanced in vitro platforms.

Purpose of the Study:

  • To introduce a computer-controlled ex vivo mouse hindlimb culturing platform for dynamic mechanical loading.
  • To evaluate muscle regeneration in a critical-size tibialis anterior defect treated with a myoblast-laden scaffold.
  • To assess the combined effects of mechanical stimulation and a polycaprolactone/collagen scaffold on muscle repair.

Main Methods:

  • Utilized an ex vivo mouse hindlimb culture system capable of applying physiological strain.
  • Created an injectable scaffold (PNCOL) from polycaprolactone nanofibers and collagen, loaded with myoblasts.
  • Treated critical-size tibialis anterior defects with the PNCOL scaffold under dynamic mechanical loading.

Main Results:

  • The ex vivo platform maintained tissue viability and structural integrity.
  • PNCOL treatment with mechanical loading improved muscle fiber organization and extracellular matrix regeneration.
  • Upregulation of key myogenic regulatory factors (PAX7, MYF5, MRF4) and TGFβ1 observed.
  • Enhanced anti-inflammatory response (CD206) and an anabolic shift in WNT and IGF-1 signaling pathways.

Conclusions:

  • The developed ex vivo platform offers a reproducible and ethical model for musculoskeletal regenerative research.
  • Combined mechanical stimulation and PNCOL biomaterial therapy significantly promotes muscle regeneration.
  • This approach advances the development of therapies for volumetric muscle loss injuries while reducing animal use.