The Role of Nigella Sativa and Phyllanthus Urinaria L Extracts Enhance Inflammation Cytokine and Growth factor in Mesenchymal Stem Cells Conditioned Medium
Keywords:
Mesenchymal Stem Cells, Conditioned Medium, Nigella sativa, Phyllanthus urinaria
Abstract
Background: Mesenchymal Stem Cells (MSCs) are cells that have the multipotent ability to undergo self-renewal, differentiate and secrete various bioactive substances, such as chemokines, proteins, microRNAs (miRNAs), growth factors, and cytokines. Conditioned medium of MSC is a medium resulting from cell culture enriched with the secretome of the cultured cells. MSC-CM treated with certain factors can increase the production of growth factors such as VEGF and PDGF, which play a role in angiogenesis and tissue repair. Modification of MSC-CM with bioactive compounds can be a promising strategy to increase the effectiveness of MSCs in medical therapy. Therefore, this study aims to examine whether these herbal extracts can modulate the production of pro-inflammatory cytokines (TNF-α, IFN-γ, IL-1β, IL-6) and growth factors (SDF-1, PDGF, VEGF) in MSC conditioned medium. Methods: This study used a pre-post research design with four treatment groups. Medium culture of MSCs treated with Nigella sativa (doses of 10 µg/mL) and Phyllanthus urinaria L (doses of 25 µg/mL), which were cultured for 24 and 48 hours. Measurement of cytokine and growth factor levels was carried out using the Enzyme-Linked Immunosorbent Assay (ELISA) method for quantitative analysis. Results: The data reveal distinct patterns in the modulation of protein levels, particularly for SDF-1, IL-1β, IL-6, TNF-α, and IFN-γ, which are critical players in inflammation and tissue regeneration. Conclusion: This study showed that the Nigella sativa and Phyllanthus urinaria L extracts in modifying conditioned medium of Mesenchymal Stem Cells is significant release of pro-inflammatory cytokines and growth factors.
References
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31. Karki, R., Sharma, B. R., Tuladhar, S., Williams, E. P., Zalduondo, L., Samir, P., Zheng, M., Sundaram, B., Banoth, B., Malireddi, R. K. S., Schreiner, P., Neale, G., Vogel, P., Webby, R., Jonsson, C. B., & Kanneganti, T. D. (2021). Synergism of TNF-α and IFN-γ Triggers Inflammatory Cell Death, Tissue Damage, and Mortality in SARS-CoV-2 Infection and Cytokine Shock Syndromes. Cell, 184(1), 149–168.e17. https://doi.org/10.1016/j.cell.2020.11.025
32. Ciesielska-Figlon, K., Wojciechowicz, K., Wardowska, A., & Lisowska, K. A. (2023). The immunomodulatory effect of Nigella sativa. Antioxidants, 12(7), 1340. https://doi.org/10.3390/antiox12071340
33. Jantan, I., Haque, M. A., Ilangkovan, M., & Arshad, L. (2019). An Insight Into the Modulatory Effects and Mechanisms of Action of Phyllanthus Species and Their Bioactive Metabolites on the Immune System. Frontiers in pharmacology, 10, 878. https://doi.org/10.3389/fphar.2019.00878
34. Yang, Y. K., Ogando, C. R., Wang See, C., Chang, T. Y., & Barabino, G. A. (2018). Changes in phenotype and differentiation potential of human mesenchymal stem cells aging in vitro. Stem cell research & therapy, 9(1), 131. https://doi.org/10.1186/s13287-018-0876-3
35. Fonseca, L. N., Bolívar-Moná, S., Agudelo, T., Beltrán, L. D., Camargo, D., Correa, N., Del Castillo, M. A., Fernández de Castro, S., Fula, V., García, G., Guarnizo, N., Lugo, V., Martínez, L. M., Melgar, V., Peña, M. C., Pérez, W. A., Rodríguez, N., Pinzón, A., Albarracín, S. L., … Gutiérrez-Gómez, M. L. (2023). Cell surface markers for mesenchymal stem cells related to the skeletal system: A scoping review. Heliyon, 9(2). https://doi.org/10.1016/j.heliyon.2023.e13464
2. Rosochowicz, M.A., Lach, M.S., Richter, M. et al. Conditioned Medium – Is it an Undervalued Lab Waste with the Potential for Osteoarthritis Management?. Stem Cell Rev and Rep 19, 1185–1213 (2023). https://doi.org/10.1007/s12015-023-10517-1
3. Hadi V, Kheirouri S, Alizadeh M, Khabbazi A, Hosseini H. Effects of Nigella sativa oil extract on inflammatory cytokine response and oxidative stress status in patients with rheumatoid arthritis: a randomized, double-blind, placebo-controlled clinical trial. Avicenna J Phytomed. 2016;6(1):34-43.
4. Lai, H. Y., Chen, Y. J., Mersmann, H. J., Lin, Y. Y., & Ding, S. T. (2024). Phyllanthus urinaria water extract ameliorates lipid accumulation, oxidative stress and inflammation in chickens with fatty liver syndrome. Italian Journal of Animal Science, 23(1), 1233–1249. https://doi.org/10.1080/1828051X.2024.2383313
5. Faraj SS, Jalal PJ. IL1β, IL-6, and TNF-α cytokines cooperate to modulate a complicated medical condition among COVID-19 patients: case-control study. Ann Med Surg (Lond). 2023;85(6):2291-2297. Published 2023 Apr 26. doi:10.1097/MS9.0000000000000679
6. Liu, Jie & Huang, Xiajie & Su, Hongjie & Yu, Jie & Nie, Xinyu & Liu, Kaibing & Qin, Wencong & Zhao, Yongxin & Su, Yongfeng & Kuang, Xiaocong & Chen, Di & Lu, William & Chen, Yan & Hua, Qikai. (2024). Tibial Cortex Transverse Transport Facilitates Severe Diabetic Foot Wound Healing via HIF-1α-Induced Angiogenesis. Journal of Inflammation Research. 17. 2681-2696. 10.2147/JIR.S456590.
7. Spelman, Kevin & Burns, Jj & Nichols, Douglas & Winters, Nasha & Ottersberg, Steve & Tenborg, Mark. (2006). Modulation of cytokine expression by traditional medicines: A review of herbal immunomodulators. Alternative medicine review : a journal of clinical therapeutic. 11. 128-50.
8. Barros S, et al. Mesenchymal stem cells: from basic science to clinical applications. Curr Stem Cell Res Ther. 2017;12(6):495-508.
9. Zhang Y, et al. Mesenchymal stem cell-derived conditioned medium: an emerging therapeutic strategy for tissue repair. Regener Med. 2015;10(1):77-89.
10. El-Mahdy MA, et al. Thymoquinone and its potential therapeutic role in inflammatory diseases. J Inflamm Res. 2019;12:69-81.
11. Matsuda H, et al. Phytochemical studies on Phyllanthus urinaria L. and its immunomodulatory effects. J Ethnopharmacol. 2004;91(2-3):287-292.
12. Tarkowski M, et al. Growth factors in stem cell therapy: SDF-1, PDGF, VEGF. Stem Cells Transl Med. 2014;3(3):303-311.
13. Ali BH, et al. The pharmacological effects of Nigella sativa L. and its active constituent, thymoquinone. Phytother Res. 2016;30(9):1461-1470.
14. Al-Majed AR, et al. The role of Nigella sativa in tissue repair and wound healing: an experimental study in rats. J Ethnopharmacol. 2015;172:174-182.
15. Dominici M, et al. Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy. 2006;8(4):315-317.
16. El-Mahdy MA, et al. Thymoquinone and its potential therapeutic role in inflammatory diseases. J Inflamm Res. 2019;12:69-81.
17. Lai PH, et al. Mesenchymal stem cells: biological characteristics and applications in regenerative medicine. J Biomed Sci. 2010;17:2.
18. Liang YY, et al. Phyllanthus urinaria L. extract modulates mesenchymal stem cell differentiation and wound healing. J Ethnopharmacol. 2018;217:132-141.
19. Matsuda H, et al. Phytochemical studies on Phyllanthus urinaria L. and its immunomodulatory effects. J Ethnopharmacol. 2004;91(2-3):287-292.
20. Prockop DJ. Mesenchymal stem/stromal cells: the state of transdifferentiation and modes of action. J Cell Mol Med. 2013;17(7):825-831.
21. Tarkowski M, et al. Growth factors in stem cell therapy: SDF-1, PDGF, VEGF. Stem Cells Transl Med. 2014;3(3):303-311.
22. Zhao, W., Jin, K., Li, J. et al. Delivery of stromal cell-derived factor 1α for in situ tissue regeneration. J Biol Eng 11, 22 (2017). https://doi.org/10.1186/s13036-017-0058-3
23. Zhao, Y., Peng, H., Yan, J., Sun, L., Huang, Y., Wei, P., Jing, W., Zhao, B., Qin, D., Liu, Y., Guo, S., Zhang, K., Wu, X., & Li, B. (2024). SDF-1α peptide-tethered SIS membrane enables biomimetic tissue regeneration via multifactorial synergetic regulation. Applied Materials Today, 39, 102293. https://doi.org/10.1016/j.apmt.2024.102293
24. Kmail, A., Said, O., & Saad, B. (2023). How Thymoquinone from Nigella sativa Accelerates Wound Healing through Multiple Mechanisms and Targets. Current issues in molecular biology, 45(11), 9039–9059. https://doi.org/10.3390/cimb45110567
25. Sallehuddin, N., Nordin, A., Bt Hj Idrus, R., & Fauzi, M. B. (2020). Nigella sativa and Its Active Compound, Thymoquinone, Accelerate Wound Healing in an In Vivo Animal Model: A Comprehensive Review. International journal of environmental research and public health, 17(11), 4160. https://doi.org/10.3390/ijerph17114160
26. Wijdan, S. A., Bokhari, S. M., Alvares, J., & Latif, V. (2025). The role of interleukin-1 beta in inflammation and the potential of immune-targeted therapies. Pharmacological Research - Reports, 3, 100027. https://doi.org/10.1016/j.prerep.2025.100027
27. Luo, Q., Hang, J., Cao, L., Wang, X., Dong, G., & Dai, W. (2024). Exploring the mechanism of Phyllanthus urinaria L. against ulcerative colitis based on network pharmacology and in vivo experiments. Chinese Journal of Analytical Chemistry, 52(12), 100465. https://doi.org/10.1016/j.cjac.2024.100465
28. Hirano T. (2021). IL-6 in inflammation, autoimmunity and cancer. International immunology, 33(3), 127–148. https://doi.org/10.1093/intimm/dxaa078
29. Aliyu, M., Zohora, F. T., Anka, A. U., Ali, K., Maleknia, S., Saffarioun, M., & Azizi, G. (2022). Interleukin-6 cytokine: An overview of the immune regulation, immune dysregulation, and therapeutic approach. International immunopharmacology, 111, 109130. https://doi.org/10.1016/j.intimp.2022.109130
30. Tanaka, T., Narazaki, M., & Kishimoto, T. (2014). IL-6 in inflammation, immunity, and disease. Cold Spring Harbor perspectives in biology, 6(10), a016295. https://doi.org/10.1101/cshperspect.a016295
31. Karki, R., Sharma, B. R., Tuladhar, S., Williams, E. P., Zalduondo, L., Samir, P., Zheng, M., Sundaram, B., Banoth, B., Malireddi, R. K. S., Schreiner, P., Neale, G., Vogel, P., Webby, R., Jonsson, C. B., & Kanneganti, T. D. (2021). Synergism of TNF-α and IFN-γ Triggers Inflammatory Cell Death, Tissue Damage, and Mortality in SARS-CoV-2 Infection and Cytokine Shock Syndromes. Cell, 184(1), 149–168.e17. https://doi.org/10.1016/j.cell.2020.11.025
32. Ciesielska-Figlon, K., Wojciechowicz, K., Wardowska, A., & Lisowska, K. A. (2023). The immunomodulatory effect of Nigella sativa. Antioxidants, 12(7), 1340. https://doi.org/10.3390/antiox12071340
33. Jantan, I., Haque, M. A., Ilangkovan, M., & Arshad, L. (2019). An Insight Into the Modulatory Effects and Mechanisms of Action of Phyllanthus Species and Their Bioactive Metabolites on the Immune System. Frontiers in pharmacology, 10, 878. https://doi.org/10.3389/fphar.2019.00878
34. Yang, Y. K., Ogando, C. R., Wang See, C., Chang, T. Y., & Barabino, G. A. (2018). Changes in phenotype and differentiation potential of human mesenchymal stem cells aging in vitro. Stem cell research & therapy, 9(1), 131. https://doi.org/10.1186/s13287-018-0876-3
35. Fonseca, L. N., Bolívar-Moná, S., Agudelo, T., Beltrán, L. D., Camargo, D., Correa, N., Del Castillo, M. A., Fernández de Castro, S., Fula, V., García, G., Guarnizo, N., Lugo, V., Martínez, L. M., Melgar, V., Peña, M. C., Pérez, W. A., Rodríguez, N., Pinzón, A., Albarracín, S. L., … Gutiérrez-Gómez, M. L. (2023). Cell surface markers for mesenchymal stem cells related to the skeletal system: A scoping review. Heliyon, 9(2). https://doi.org/10.1016/j.heliyon.2023.e13464
Published
2025-04-26
How to Cite
Ayu, D. R., Irawan, R. C. S., Prawitasari, S., & Shindy, M. (2025). The Role of Nigella Sativa and Phyllanthus Urinaria L Extracts Enhance Inflammation Cytokine and Growth factor in Mesenchymal Stem Cells Conditioned Medium . International Journal of Cell and Biomedical Science, 3(7), 201-208. https://doi.org/10.59278/cbs.v3i7.48
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