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Purpose <p>The aim of this study is to investigate the role of extracellular vesicles (EVs) derived from <i>Perionyx excavatus</i> in tissue repair and regeneration, highlighting their potential application in regenerative medicine.</p> Methods <p>EVs were isolated from distinct regions of <i>P. excavatus</i> using ultracentrifugation and characterised by advanced imaging and analytical techniques, including scanning electron microscopy (SEM), transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA), and dynamic light scattering (DLS). Gene expression analysis was conducted to evaluate the differential miRNA and mRNA profiles. Functional assays, including wound-healing and real-time cell confluency analyses, were performed to assess the regenerative potential of EVs on mouse fibroblast cells (NIH-3T3).</p> Results <p>The isolated EVs were confirmed to exhibit sizes and morphologies comparable to EVs derived from other organisms. Gene expression analysis revealed that blastema EVs possess significantly higher levels of regenerative markers, such as pex-miR-365 and Sox-C, compared to anterior segment EVs. Functional assays have shown efficient internalisation of EVs by NIH-3T3 cells, with fluorescence intensities for anterior segment EVs at 14.2% and blastema EVs at 34% compared to the control. Enhanced cell proliferation and migration were observed in EV-treated groups, as evidenced by significant improvements in wound closure and real-time cell confluency analysis.</p> Conclusion <p>This study demonstrates the therapeutic value of <i>P. excavatus-</i>derived EVs, driven by their unique miRNA and mRNA expression profiles. These findings provide a foundation for the development of EV-based therapies in regenerative medicine, offering promising avenues for therapeutic innovation.</p> Lay Summary <p>EVs are tiny bioactive carriers secreted by various organisms, rich in RNA, proteins, and enzymes. In this study, EVs isolated from <i>P. excavatus</i> were evaluated for their role in tissue repair and regeneration. Using advanced imaging and gene expression analysis, EVs isolated from regenerative segments of the earthworm exhibit higher levels of regenerative-specific markers. Functional studies further demonstrated that these EVs enhance cell growth and migration, making them a promising tool in regenerative medicine.</p> Graphical Abstract <p></p>

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Regenerating Earthworm-Derived Extracellular Vesicles Enhance Mouse Fibroblast Growth and Migration

  • 期刊论文

  • 发表日期：

    2026年2月2日

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  • Ravichandran Subramaniam,
  • Johnson Retnaraj Samuel Selvan Christyraj,
  • Melinda Grace Rossan Mathews,
  • Saravanakumar Venkatachalam,
  • Beryl Vedha Yesudhason

摘要

Purpose

The aim of this study is to investigate the role of extracellular vesicles (EVs) derived from Perionyx excavatus in tissue repair and regeneration, highlighting their potential application in regenerative medicine.

Methods

EVs were isolated from distinct regions of P. excavatus using ultracentrifugation and characterised by advanced imaging and analytical techniques, including scanning electron microscopy (SEM), transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA), and dynamic light scattering (DLS). Gene expression analysis was conducted to evaluate the differential miRNA and mRNA profiles. Functional assays, including wound-healing and real-time cell confluency analyses, were performed to assess the regenerative potential of EVs on mouse fibroblast cells (NIH-3T3).

Results

The isolated EVs were confirmed to exhibit sizes and morphologies comparable to EVs derived from other organisms. Gene expression analysis revealed that blastema EVs possess significantly higher levels of regenerative markers, such as pex-miR-365 and Sox-C, compared to anterior segment EVs. Functional assays have shown efficient internalisation of EVs by NIH-3T3 cells, with fluorescence intensities for anterior segment EVs at 14.2% and blastema EVs at 34% compared to the control. Enhanced cell proliferation and migration were observed in EV-treated groups, as evidenced by significant improvements in wound closure and real-time cell confluency analysis.

Conclusion

This study demonstrates the therapeutic value of P. excavatus-derived EVs, driven by their unique miRNA and mRNA expression profiles. These findings provide a foundation for the development of EV-based therapies in regenerative medicine, offering promising avenues for therapeutic innovation.

Lay Summary

EVs are tiny bioactive carriers secreted by various organisms, rich in RNA, proteins, and enzymes. In this study, EVs isolated from P. excavatus were evaluated for their role in tissue repair and regeneration. Using advanced imaging and gene expression analysis, EVs isolated from regenerative segments of the earthworm exhibit higher levels of regenerative-specific markers. Functional studies further demonstrated that these EVs enhance cell growth and migration, making them a promising tool in regenerative medicine.

Graphical Abstract