Antibacterial and Cytotoxic Activity of Silver and Zinc Oxide Nanoparticles Synthesized Using Deep Eutectic Solvent Extraction of the Sea Cucumber-Holothuria tubulosa

Year 2025, Volume: 18 Issue: 3, 745 - 762

Hilal Top-haytaoğlu , Ayşegül İnam , Tülay Öncü Öner , Murat Elibol

Abstract

Silver nanoparticles (AgNPs) and zinc oxide nanoparticles (ZnO-NPs) have garnered significant attention for their biomedical applications, owing to their potent antibacterial properties and cytotoxicity against cancer cells. In this study, we synthesized these nanoparticles (NPs) utilizing a novel approach involving deep eutectic solvent (DES) extraction of the sea cucumber-Holothuria tubulosa. Choline chloride-urea (1:2) was used as a DES, facilitating the green synthesis without hazardous chemicals. Spectrophotometric methods were used to determine total phenolic and carbohydrate content of the extract. UV–Vis, DLS and FTIR characterized NPs. The absorption peaks in 400 nm for AgNPs and 378 nm for ZnO-NPs were observed on the UV-Vis. DLS prediction of AgNPs size at 202.2 nm and ZnO-NPs size at 269.9 nm. The cytotoxic activities of the synthesized NPs and DES extract of H. tubulosa were evaluated against SH-SY5Y neuroblastoma and BJ normal skin fibroblast cell lines using MTT assay. Cytotoxic effects were observed for all samples in SH-SY5Y cells. However, the IC50 value could not be calculated for BJ cells because it was outside the range of the highest concentration tested. Moreover, the antibacterial activities of NPs and DES extract of H. tubulosa were investigated against Escherichia coli and Staphylococcus aureus. The AgNPs exhibited significant inhibitory effects against both tested bacterial strains, demonstrating their antibacterial activity. Overall, the green synthesis of NPs using DES derived from sea cucumber represents a sustainable approach. These findings underscore potential of the sea cucumber-derived DES-mediated synthesis of NPs for biomedical applications, including cancer therapy and antimicrobial coatings.

Keywords

Deep eutectic solvents , sea cucumber , nanoparticles , antibacterial , cytotoxicity

References

• [1] Fagbohun, O. F., Joseph, J. S., Oriyomi, O. V., & Rupasinghe, H. V. (2023). Saponins of North Atlantic Sea Cucumber: Chemistry, health benefits, and future prospectives. Marine Drugs, 21(5), 262.
• [2] Maskur, M., Sayuti, M., Widyasari, F., & Setiarto, R. H. B. (2024). Bioactive Compound and Functional Properties of Sea Cucumbers as Nutraceutical Products. Reviews in Agricultural Science, 12, 45-64.
• [3] Sanjeewa, K. K., & Herath, K. H. I. N. M. (2023). Bioactive secondary metabolites in sea cucumbers and their potential to use in the functional food industry. Fisheries and Aquatic Sciences, 26(2), 69-86.
• [4] Liu, J., Li, X., & Row, K. H. (2022). Development of deep eutectic solvents for sustainable chemistry. Journal of Molecular Liquids, 362, 119654.
• [5] Liu, Y., Friesen, J. B., McAlpine, J. B., Lankin, D. C., Chen, S. N., & Pauli, G. F. (2018). Natural deep eutectic solvents: properties, applications, and perspectives. Journal of natural products, 81(3), 679-690.
• [6] Dubois, M., Gilles, K. A., Hamilton, J. K., Rebers, P. T., & Smith, F. (1956). Colorimetric method for determination of sugars and related substances. Analytical chemistry, 28(3), 350-356.
• [7] Singleton, V. L., Orthofer, R., & Lamuela-Raventós, R. M. (1999). Analysis of total phenols and other oxidation substrates and antioxidants by means of folin-ciocalteu reagent. In Methods in enzymology, 299, 152.
• [8] Tüzün, S., Baş, İ., Karakavuk, E., Sanyürek, N. K., & Benzer, F. (2020). Çeşitli pekmez türlerinde farklı yöntemlerle tespit edilen antioksidan aktivitelerin karşılaştırılması. Türk Tarım ve Doğa Bilimleri Dergisi, 7(2), 323-330.
• [9] Matuschek, E., Copsey-Mawer, S., Petersson, S., Åhman, J., Morris, T. E., & Kahlmeter, G. (2023). The European committee on antimicrobial susceptibility testing disc diffusion susceptibility testing method for frequently isolated anaerobic bacteria. Clinical Microbiology and Infection, 29(6), 795-e1.
• [10] Khanavi, M., Gheidarloo, R., Sadati, N., Ardekani, M. R. S., Nabavi, S. M. B., Tavajohi, S., Ostad, S. N. (2012). Cytotoxicity of fucosterol containing fraction of marine algae against breast and colon carcinoma cell line. Pharmacognosy Magazine, 8(29), 60.
• [11] Wang, J., Shi, S., Li, F., Du, X., Kong, B., Wang, H., & Xia, X. (2022). Physicochemical properties and antioxidant activity of polysaccharides obtained from sea cucumber gonads via ultrasound-assisted enzymatic techniques,160, 113307.
• [12] Srikar, S. K., Giri, D. D., Pal, D. B., Mishra, P. K., & Upadhyay, S. N. (2016). Green synthesis of silver nanoparticles: a review. Green and Sustainable Chemistry, 6(01), 34.
• [13] Thanigaivel, S., Vickram, S., Saranya, V., Ali, H., Alarifi, S., Modigunta, J. K. R., Rohini, K. (2022). Seaweed polysaccharide mediated synthesis of silver nanoparticles and its enhanced disease resistance in Oreochromis mossambicus. Journal of King Saud University-Science, 34(2), 101771.
• [14] Nurkhasanah, N., Yuwono, T., Nurani, L. H., Rizki, M. I., & Kraisintu, K. (2015). The development of chitosan nanoparticles from Hibiscus sabdariffa L calyx extract from Indonesia and Thailand, 6(5), 1855-1861.
• [15] Sivasankar, M., & Kumar, B. P. (2010). Role of nanoparticles in drug delivery system. International Journal of Research in Pharmaceutical and Biomedical Sciences, 1(2), 41-66.
• [16] Mohammadian, M., Es’haghi, Z., & Hooshmand, S. (2018). Green and chemical synthesis of zinc oxide nanoparticles and size evaluation by UV–vis spectroscopy. J Nanomed Res, 7(1), 00175.
• [17] Fakhari, S., Jamzad, M., & Kabiri Fard, H. (2019). Green synthesis of zinc oxide nanoparticles: a comparison. Green chemistry letters and reviews, 12(1), 19-24.
• [18] Hasaballah, A. I., El-Naggar, H. A., Abd-El Rahman, I. E., Al-Otibi, F., Alahmadi, R. M., Abdelzaher, O. F., ... & Elbahnasawy, M. A. (2023). Surf Redfish-Based ZnO-NPs and Their Biological Activity with Reference to Their Non-Target Toxicity. Marine Drugs, 21(8), 437.
• [19] Elbahnasawy, M. A., El-Naggar, H. A., Abd-El Rahman, I. E., Kalaba, M. H., Moghannem, S. A., Al-Otibi, F., ... & Hasaballah, A. I. (2023). Biosynthesized ZnO-NPs Using Sea Cucumber (Holothuria impatiens): Antimicrobial Potential, Insecticidal Activity and In Vivo Toxicity in Nile Tilapia Fish, Oreochromis niloticus. Separations, 10(3), 173.
• [20] Jamdagni, P., Khatri, P., & Rana, J. S. (2018). Green synthesis of zinc oxide nanoparticles using flower extract of Nyctanthes arbor-tristis and their antifungal activity. Journal of King Saud University-Science, 30(2), 168-175.
• [21] Rosman, N. S. R., Harun, N. A., Idris, I., & Ismail, W. I. W. (2020). Eco-friendly silver nanoparticles (AgNPs) fabricated by green synthesis using the crude extract of marine polychaete, Marphysa moribidii: biosynthesis, characterisation, and antibacterial applications. Heliyon, 6(11).
• [22] Willian, N., Syukri, S., Zulhadjri, Z., Pardi, H., & Arief, S. (2021). Marine plant mediated green synthesis of silver nanoparticles using mangrove Rhizophora stylosa: Effect of variable process and their antibacterial activity, F1000Research, 10.
• [23] Doğangüneş, M., Türker, A., Güneş, H., & Alper, M. (2021). Cytotoxic and Antioxidant Potential of Chloroform Extract of Holothuria tubulosa Gmelin, 1791. Russian Journal of Marine Biology, 47(6), 498-507.
• [24] Alper, M., & Güneş, M. (2020). Evaluation of cytotoxic, apoptotic effects and phenolic compounds of sea cucumber Holothuria tubulosa (Gmelin, 1791) extracts. Turkish Journal of Veterinary & Animal Sciences, 44(3), 641-655.
• [25] Karmous, I., Pandey, A., Haj, K. B., & Chaoui, A. (2020). Efficiency of the green synthesized nanoparticles as new tools in cancer therapy: insights on plant-based bioengineered nanoparticles, biophysical properties, and anticancer roles. Biological trace element research, 196, 330-342.