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Related Experiment Video

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Novel Three-Dimensional Preclinical Model for Investigating Cartilage Regeneration, Incorporating Physiological and

Mingjing Zhu1, Jianfeng Jin1, Hadi Seddiqi1

  • 1Department of Oral Cell Biology, Academic Centre for Dentistry Amsterdam (ACTA), Amsterdam Movement Sciences, University of Amsterdam and Vrije Universiteit Amsterdam, Amsterdam, The Netherlands.

Tissue Engineering. Part C, Methods
|October 24, 2025
PubMed
Summary

Researchers developed a novel 3D preclinical model for temporomandibular joint disorders (TMDs) research. This in vitro system mimics in vivo conditions, enabling the study of cartilage regeneration under various mechanical loads.

Keywords:
3D in vitro modelchondrogenic differentiationcompressive mechanical loadingconstant loadingdynamic loading

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Area of Science:

  • Biomaterials Science
  • Tissue Engineering
  • Orthopedics

Background:

  • Current temporomandibular joint disorder (TMD) treatments are hindered by a lack of preclinical models that accurately replicate in vivo conditions.
  • Existing models often fail to incorporate crucial factors like three-dimensional (3D) cell culture and mechanical stimulation, which significantly impact cell behavior and treatment outcomes.
  • There is a critical need for advanced in vitro systems that can simulate physiological and pathological mechanical loading relevant to TMDs.

Purpose of the Study:

  • To develop and validate a novel 3D in vitro preclinical model for temporomandibular joint disorders (TMDs) research.
  • To investigate the effects of dynamic moderate loading (simulating chewing) and constant excessive loading (simulating clenching) on cell behavior within the model.
  • To establish an in vivo-mimetic system for evaluating cartilage regeneration under physiologically relevant mechanical stimuli.

Main Methods:

  • Engineered cylindrical 3D constructs (3% agarose with mouse bone marrow mesenchymal stem cells - BMSCs).
  • Cultured BMSCs in chondrogenic medium to confirm chondrogenic potential (assessed via Sox9 expression).
  • Applied mechanical loading (dynamic 5% compression at 1 Hz; constant 10-30% compression) using a custom bioreactor, followed by finite element modeling analysis.

Main Results:

  • BMSCs in 3D constructs showed significantly increased Sox9 expression, confirming chondrogenic potential compared to controls.
  • Mechanical loading (dynamic and constant) elevated nitric oxide production in BMSCs.
  • Dynamic loading upregulated c-Fos and c-Jun expression, while constant loading increased c-Jun expression; excessive constant loading (30%) compromised construct integrity and 20% constant strain reduced live cell count.

Conclusions:

  • A novel 3D in vitro preclinical model simulating in vivo mechanical conditions for TMDs has been successfully established.
  • The model allows for the investigation of cartilage regeneration under physiological and pathological mechanical stimuli.
  • This platform provides a promising tool for future research on chondroinductive agents and mechanical loading therapies for TMDs.