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Tracking chondrocyte-to-fibroblast transformation via changes in cell electrophysiology
Krista S P Clarke1, Rebecca Lewis2, Michael Pycraft Hughes1,3
1Centre for Biomedical Engineering, School of Mechanical Engineering, University of Surrey, Guildford, Surrey, GU2 7XH, UK.
Scientific Reports
|May 7, 2025
Summary
Dielectrophoresis (DEP) monitors electrophysiological changes in cultured chondrocytes, revealing cytoplasmic conductivity shifts over 100 days. This method reliably tracks dedifferentiation from chondrocytes to fibroblasts, crucial for cartilage regeneration therapies.
Area of Science:
- Biomedical Engineering
- Cell Biology
- Tissue Engineering
Background:
- Cultured chondrocytes are vital for cartilage regeneration but dedifferentiate in monolayer culture, impacting clinical applications.
- Chondrocyte dedifferentiation to a fibroblastic phenotype hinders effective cell-based cartilage repair, leading to suboptimal outcomes.
Purpose of the Study:
- To investigate the use of dielectrophoresis (DEP) for monitoring electrophysiological changes in cultured chondrocytes.
- To establish cytoplasmic conductivity monitoring as a reliable method for tracking chondrocyte dedifferentiation over extended culture periods.
Main Methods:
- Primary bovine chondrocytes were cultured and their electrophysiological properties analyzed using DEP over 100 days.
- A multi-conductivity approach was employed to measure cytoplasmic conductivity and membrane capacitance.
- Cellular transitions were tracked by analyzing cytoplasmic vs. medium conductivity to determine membrane potential (Vm).
Main Results:
- Statistically significant changes in membrane capacitance (p=0.0039) and cytoplasmic conductivity (p<0.0001) were observed.
- Cytoplasmic conductivity monitoring reliably tracked chondrocyte dedifferentiation over 100 days in culture.
- Membrane potential transitions indicated shifts from chondrocytes (-13 to -18 mV) to proliferating (-32 to -43 mV) and non-proliferating fibroblasts (-55 to -71 mV) around days 40 and 80.
Conclusions:
- Dielectrophoresis offers a reliable method for tracking chondrocyte dedifferentiation in vitro.
- Monitoring cytoplasmic conductivity and membrane potential provides insights into cell state changes relevant to cartilage regeneration.
- This technique can aid in optimizing cell culture conditions and improving outcomes for cartilage repair therapies.
