Enhancing the wound healing potential using earthworm clitellum factors and elucidating its molecular mechanism in an in-vitro and earthworm model
Abstract Earthworm, Eudrilus eugeniae cannot survive and regenerate without clitellum segments. In regenerating worms, the clitellum’s epithelial and circular muscular layers are reduced to one-third, and longitudinal cell layers to half. In C2C12 cells, Clitellum Factors (CF − 5, 25 and 50%) and Re...
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Nature Portfolio
2024-11-01
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| Online Access: | https://doi.org/10.1038/s41598-024-79304-3 |
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| author | Kamarajan Rajagopalan Jackson Durairaj Selvan Christyraj Johnson Retnaraj Samuel Selvan Christyraj Meikandan Chandrasekar Nivedha Balamurugan Nandha Kumar Suresh Puja Das Ashwin Barath Vaidhyalingham Leela Bharathiraja |
| author_facet | Kamarajan Rajagopalan Jackson Durairaj Selvan Christyraj Johnson Retnaraj Samuel Selvan Christyraj Meikandan Chandrasekar Nivedha Balamurugan Nandha Kumar Suresh Puja Das Ashwin Barath Vaidhyalingham Leela Bharathiraja |
| author_sort | Kamarajan Rajagopalan |
| collection | DOAJ |
| description | Abstract Earthworm, Eudrilus eugeniae cannot survive and regenerate without clitellum segments. In regenerating worms, the clitellum’s epithelial and circular muscular layers are reduced to one-third, and longitudinal cell layers to half. In C2C12 cells, Clitellum Factors (CF − 5, 25 and 50%) and Regenerative Clitellum Factors (RCF − 5, 25, 50, 75%) ameliorate the cell viability up to 20–28% and 30–38% respectively than the control. In contrast, extracts from body segments negatively influence cell viability up to 80%. In a scratch-wound assay, 25% RCF and 5% CF achieved 99.86% and 81.54% wound closure in 24 h, respectively, compared to 40% in controls. RCF and CF also possess enhanced anti-microbial activity against gram + ve bacteria. Western Blotting reveals that Wnt3a, HoxD3 and VEGF were remarkably upregulated in RCF and CF treated samples and their upregulated stemness property is effectively regulated by p53, TCTP, H2AX, Cleaved Caspase-3 proteins. Immunofluorescence data clearly states that Wnt3a and Caspase-3 signals are more profoundly observed in nuclear over cytoplasm in RCF treated samples and H2AX shows less nuclear signals than CF. In in-vivo earthworm model conditions, RCF remarkably promotes the survivability and wound healing ability by promoting the Wnt3a and VEGF expression together with downregulation of Cox2. |
| format | Article |
| id | doaj-art-08a2da56d5bb46f39efdc17f9edb6878 |
| institution | Kabale University |
| issn | 2045-2322 |
| language | English |
| publishDate | 2024-11-01 |
| publisher | Nature Portfolio |
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| series | Scientific Reports |
| spelling | doaj-art-08a2da56d5bb46f39efdc17f9edb68782024-11-17T12:25:43ZengNature PortfolioScientific Reports2045-23222024-11-0114111610.1038/s41598-024-79304-3Enhancing the wound healing potential using earthworm clitellum factors and elucidating its molecular mechanism in an in-vitro and earthworm modelKamarajan Rajagopalan0Jackson Durairaj Selvan Christyraj1Johnson Retnaraj Samuel Selvan Christyraj2Meikandan Chandrasekar3Nivedha Balamurugan4Nandha Kumar Suresh5Puja Das6Ashwin Barath Vaidhyalingham7Leela Bharathiraja8Molecular Biology and Stem Cell Research Lab, Centre for Molecular and Nanomedical Sciences, International Research Centre, Sathyabama Institute of Science & Technology (Deemed to be University)Molecular Biology and Stem Cell Research Lab, Centre for Molecular and Nanomedical Sciences, International Research Centre, Sathyabama Institute of Science & Technology (Deemed to be University)Molecular Biology and Stem Cell Research Lab, Centre for Molecular and Nanomedical Sciences, International Research Centre, Sathyabama Institute of Science & Technology (Deemed to be University)Molecular Biology and Stem Cell Research Lab, Centre for Molecular and Nanomedical Sciences, International Research Centre, Sathyabama Institute of Science & Technology (Deemed to be University)Molecular Biology and Stem Cell Research Lab, Centre for Molecular and Nanomedical Sciences, International Research Centre, Sathyabama Institute of Science & Technology (Deemed to be University)Jaya College of Arts and ScienceMolecular Biology and Stem Cell Research Lab, Centre for Molecular and Nanomedical Sciences, International Research Centre, Sathyabama Institute of Science & Technology (Deemed to be University)Molecular Biology and Stem Cell Research Lab, Centre for Molecular and Nanomedical Sciences, International Research Centre, Sathyabama Institute of Science & Technology (Deemed to be University)Molecular Biology and Stem Cell Research Lab, Centre for Molecular and Nanomedical Sciences, International Research Centre, Sathyabama Institute of Science & Technology (Deemed to be University)Abstract Earthworm, Eudrilus eugeniae cannot survive and regenerate without clitellum segments. In regenerating worms, the clitellum’s epithelial and circular muscular layers are reduced to one-third, and longitudinal cell layers to half. In C2C12 cells, Clitellum Factors (CF − 5, 25 and 50%) and Regenerative Clitellum Factors (RCF − 5, 25, 50, 75%) ameliorate the cell viability up to 20–28% and 30–38% respectively than the control. In contrast, extracts from body segments negatively influence cell viability up to 80%. In a scratch-wound assay, 25% RCF and 5% CF achieved 99.86% and 81.54% wound closure in 24 h, respectively, compared to 40% in controls. RCF and CF also possess enhanced anti-microbial activity against gram + ve bacteria. Western Blotting reveals that Wnt3a, HoxD3 and VEGF were remarkably upregulated in RCF and CF treated samples and their upregulated stemness property is effectively regulated by p53, TCTP, H2AX, Cleaved Caspase-3 proteins. Immunofluorescence data clearly states that Wnt3a and Caspase-3 signals are more profoundly observed in nuclear over cytoplasm in RCF treated samples and H2AX shows less nuclear signals than CF. In in-vivo earthworm model conditions, RCF remarkably promotes the survivability and wound healing ability by promoting the Wnt3a and VEGF expression together with downregulation of Cox2.https://doi.org/10.1038/s41598-024-79304-3Wound healingClitellum factorsC2C12Wnt3aVEGFCox2 |
| spellingShingle | Kamarajan Rajagopalan Jackson Durairaj Selvan Christyraj Johnson Retnaraj Samuel Selvan Christyraj Meikandan Chandrasekar Nivedha Balamurugan Nandha Kumar Suresh Puja Das Ashwin Barath Vaidhyalingham Leela Bharathiraja Enhancing the wound healing potential using earthworm clitellum factors and elucidating its molecular mechanism in an in-vitro and earthworm model Scientific Reports Wound healing Clitellum factors C2C12 Wnt3a VEGF Cox2 |
| title | Enhancing the wound healing potential using earthworm clitellum factors and elucidating its molecular mechanism in an in-vitro and earthworm model |
| title_full | Enhancing the wound healing potential using earthworm clitellum factors and elucidating its molecular mechanism in an in-vitro and earthworm model |
| title_fullStr | Enhancing the wound healing potential using earthworm clitellum factors and elucidating its molecular mechanism in an in-vitro and earthworm model |
| title_full_unstemmed | Enhancing the wound healing potential using earthworm clitellum factors and elucidating its molecular mechanism in an in-vitro and earthworm model |
| title_short | Enhancing the wound healing potential using earthworm clitellum factors and elucidating its molecular mechanism in an in-vitro and earthworm model |
| title_sort | enhancing the wound healing potential using earthworm clitellum factors and elucidating its molecular mechanism in an in vitro and earthworm model |
| topic | Wound healing Clitellum factors C2C12 Wnt3a VEGF Cox2 |
| url | https://doi.org/10.1038/s41598-024-79304-3 |
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