Evaluation of Cytotoxicity and Antibacterial Activity of Green Synthesized Silver Nanoparticles using Hedera helix extract.

Ahmed Naji Alfalahi; sina M. Matalqah; Reem Issa; Hala I. AL-Daghistani; Anas Abed

doi:10.35516/jjps.v18i2.2620

Authors

Ahmed Naji Alfalahi Department of pharmaceutics and technology, Faculty of pharmacy, Pharmacological and Diagnostics Research Center (PDRC), Faculty of Pharmacy, AL-Ahliyya Amman University, Jordan.
sina M. Matalqah Department of pharmaceutics and technology, Faculty of pharmacy, Pharmacological and Diagnostics Research Center (PDRC), Faculty of Pharmacy, AL-Ahliyya Amman University, Jordan.
Reem Issa Department of pharmaceutical Science, Faculty of pharmacy, Pharmacological and Diagnostics Research Center (PDRC), Faculty of Pharmacy, AL-Ahliyya Amman University, Jordan.
Hala I. AL-Daghistani Department of Medical Laboratory Science, Faculty of Allied Medical Science, Amman, Jordan.
Anas Abed Department of biopharmaceutics and clinical pharmacy, Faculty of pharmacy, Pharmacological and Diagnostics Research Center (PDRC), Faculty of Pharmacy, AL-Ahliyya Amman University, Jordan.

DOI:

https://doi.org/10.35516/jjps.v18i2.2620

Keywords:

Hedera helix, Silver Nanoparticles, Green synthesis, S. aureus, P. aeruginosa, A549 lung cancer cell lines

Abstract

Nowadays, silver nanoparticles (AgNPs) have drawn significant interest due to their unique properties, making them advantageous in biomedical applications, sensors, antimicrobial agents, catalysts, and optical fibers. Green synthesis is the safest and easiest method for producing silver nanoparticles (AgNPs). This study aimed to investigate the antibacterial and cytotoxic activities of silver nanoparticles synthesized using aqueous extracts of Hedera helix (AHE) against S. aureus, P. aeruginosa, and A549 lung cancer cell lines. Silver nanoparticles (Hh-AgNPs) were synthesized using the aqueous extracts of Hedera helix (AHE) as a reducing agent and polyvinylpyrrolidone (PVP) as a stabilizer and characterized by UV-visible spectrophotometry and particle size analysis via dynamic light scattering (DLS). The silver nanoparticles (Hh-AgNPs) were successfully synthesized, showing maximum absorption at 448 nm, with enhanced cytotoxic activity against A549 lung cancer cell lines (IC₅₀ = 15.16 µg/ml) and antibacterial activity against S. aureus (MIC = 0.156 mg/ml) and P. aeruginosa (MIC = 0.3125 mg/ml), compared to AHE alone. Biological methods are cost-effective and eco-friendly and thus can serve as an economical and efficient alternative for the large-scale synthesis of silver nanoparticles.

References

Khan I., Saeed K. and Khan I. Nanoparticles: Properties, applications and toxicities. Arabian J. Chem. 2019; 12(7):908-931. DOI: https://doi.org/10.1016/j.arabjc.2017.05.011

Matalqah S., Lafi Z. and Al-Kabariti A.Y. A recent review of PLGA-PEG hybrid nanoparticles for anticancer and anti-inflammatory applications. Jordan J. Pharm. Sci. 2025; 18(1):180-195. DOI: https://doi.org/10.35516/jjps.v18i1.2737

Iravani S., Korbekandi H., Mirmohammadi S.V. and Zolfaghari B. Synthesis of silver nanoparticles: chemical, physical and biological methods. Res. Pharm. Sci. 2014; 9(6):385-406.

Ghorbani H.R., Safekordi A.A., Attar H. and Sorkhabadi S.M. Biological and non-biological methods for silver nanoparticles synthesis. Chem. Biochem. Eng. Q. 2011; 25(3):317-326.

Vithiya K. and Sen S. Biosynthesis of nanoparticles. Int. J. Pharm. Sci. Res. 2011; 2(11):2781.

Patel S. A review on synthesis of silver nanoparticles-a green expertise. Life Sci. Leaflets. 2021; 132.

Ahmed S., Ahmad M., Swami B.L. and Ikram S. A review on plants extracts mediated synthesis of silver nanoparticles for antimicrobial applications: a green expertise. J. Adv. Res. 2016; 7(1):17-28. DOI: https://doi.org/10.1016/j.jare.2015.02.007

Metcalfe D.J. Hedera helix L. J. Ecol. 2005; 93(3):632-648. DOI: https://doi.org/10.1111/j.1365-2745.2005.01021.x

Al-Snafi A.E. Pharmacological and therapeutic activities of Hedera helix—a review. J. Pharm. 2018; 8(5):41-53.

Stauss-Grabo M., Atiye S., Warnke A., Wedemeyer R.S., Donath F. and Blume H.H. Observational study on the tolerability and safety of film-coated tablets containing ivy extract (Prospan® Cough Tablets) in the treatment of colds accompanied by coughing. Phytomedicine. 2011; 18(6):433-436.‏ DOI: https://doi.org/10.1016/j.phymed.2010.11.009

Fazio S., Pouso J., Dolinsky D., Fernandez A., Hernandez M., Clavier G. and Hecker M. Tolerance, safety and efficacy of Hedera helix extract in inflammatory bronchial diseases under clinical practice conditions: a prospective, open, multicentre postmarketing study in 9657 patients. Phytomedicine. 2009; 16(1):17-24. DOI: https://doi.org/10.1016/j.phymed.2006.05.003

Cwientzek U., Ottillinger B. and Arenberger P. Acute bronchitis therapy with ivy leaves extracts in a two-arm study. A double-blind, randomised study vs. another ivy leaves extract. Phytomedicine. 2011; 18(13):1105-1109. DOI: https://doi.org/10.1016/j.phymed.2011.06.014

Khan M.F., Akram M., Akhter N., Mukhtiar M., Zahid R., Khan F.S., et al. The evaluation of efficacy and safety of Cough (EMA) granules used for upper respiratory disorders. Pak. J. Pharm. Sci. 2018; 31(6):2617-2622.

Lutsenko Y.U.L.I.A., Bylka W.I.E.S.Ł.A.W.A., Matlawska I. and Darmohray R.O.M.A.N. Hedera helix as a medicinal plant. Herba Polonica. 2010; 56(1):83-96.

Mukattash H.K., Issa R., Hajleh M.N.A. and Al-Daghistani H. Inhibitory effects of polyphenols from Equisetum ramosissimum and Moringa peregrina extracts on Staphylococcus aureus, collagenase, and tyrosinase enzymes: in vitro studies. Jordan J. Pharm. Sci. 2024; 17(3):530-548. DOI: https://doi.org/10.35516/jjps.v17i3.2164

Abbasifar A., Ghani S., Irvani M.A., Rafiee B., Kaji B.V. and Akbari A. Antibacterial activity of silver nanoparticles synthesized by using extracts of Hedera helix. Zahedan J. Res. Med. Sci. 2017; 19(1). DOI: https://doi.org/10.17795/zjrms-5920

El Aziz M.M.A., Ashour A.S. and Melad A.S.G. A review on saponins from medicinal plants: chemistry, isolation, and determination. J. Nanomed. Res. 2019; 8(1):282-288. DOI: https://doi.org/10.15406/jnmr.2019.07.00199

Bissinger R., Modicano P., Alzoubi K., Honisch S., Faggio C., Abed M. and Lang F. Effect of saponin on erythrocytes. Int. J. Hematol. 2014; 100:51-59.‏ DOI: https://doi.org/10.1007/s12185-014-1605-z

Lohvina H., Sándor M. and Wink M. Effect of ethanol solvents on total phenolic content and antioxidant properties of seed extracts of Fenugreek (Trigonella foenum-graecum L.) varieties and determination of phenolic composition by HPLC-ESI-MS. Diversity. 2021; 14(1):7. DOI: https://doi.org/10.3390/d14010007

Chang C.C., Yang M.H., Wen H.M. and Chern J.C. Estimation of total flavonoid content in propolis by two complementary colorimetric methods. J. Food Drug Anal. 2002; 10(3). DOI: https://doi.org/10.38212/2224-6614.2748

Matusiewicz M., Kosieradzka I., Niemiec T., Grodzik M., Antushevich H., Strojny B. and Gołębiewska M. In vitro influence of extracts from snail Helix aspersa Müller on the colon cancer cell line Caco-2. Int. J. Mol. Sci. 2018; 19(4):1064. DOI: https://doi.org/10.3390/ijms19041064

Re R., Pellegrini N., Proteggente A., Pannala A., Yang M. and Rice-Evans C. Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Radic. Biol. Med. 1999; 26(9-10):1231-1237. DOI: https://doi.org/10.1016/S0891-5849(98)00315-3

Zein R., Alghoraibi I., Soukkarieh C., Ismail M.T. and Alahmad A. Influence of polyvinylpyrrolidone concentration on properties and anti-bacterial activity of green synthesized silver nanoparticles. Micromachines. 2022; 13(5):777. DOI: https://doi.org/10.3390/mi13050777

Cockerill F.R., Wikler M., Bush K., Dudley M., Eliopoulos G. and Hardy D. Clinical and laboratory standards institute. Performance standards for antimicrobial susceptibility testing: twenty-second informational supplement. 2012.

Abu‐Niaaj L.F., Al‐Daghistani H.I., Katampe I., Abu‐Irmaileh B. and Bustanji Y.K. Pomegranate peel: Bioactivities as antimicrobial and cytotoxic agents. Food Sci. Nutr. 2024; 12(4):2818-2832. DOI: https://doi.org/10.1002/fsn3.3963

Biemer J.J. Antimicrobial susceptibility testing by the Kirby-Bauer disc diffusion method. Ann. Clin. Lab. Sci. 1973; 3(2):135-140.‏

Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J. Immunol. Methods. 1983; 65(1-2):55-63. DOI: https://doi.org/10.1016/0022-1759(83)90303-4

Maurya D.K., Nandakumar N. and Devasagayam T.P.A. Anticancer property of gallic acid in A549, a human lung adenocarcinoma cell line, and possible mechanisms. J. Clin. Biochem. Nutr. 2010; 48(1):85-90. DOI: https://doi.org/10.3164/jcbn.11-004FR

Guta T. and Jemal K. Antioxidant and antimicrobial potentials of Nicotiana glauca Graham leaves extracts and synthesized silver nanoparticles: a phytochemical approach. Jordan J. Pharm. Sci. 2025; 18(1):57-76. DOI: https://doi.org/10.35516/jjps.v18i1.2180

Jalilian F., Chahardoli A., Sadrjavadi K., Fattahi A. and Shokoohinia Y. Green synthesized silver nanoparticle from Allium ampeloprasum aqueous extract: characterization, antioxidant activities, antibacterial and cytotoxicity effects. Adv. Powder Technol. 2020; 31(3):1323-1332. DOI: https://doi.org/10.1016/j.apt.2020.01.011

Mohanta Y.K., Panda S.K., Biswas K., Tamang A., Bandyopadhyay J., De D., et al. Biogenic synthesis of silver nanoparticles from Cassia fistula (Linn.): in vitro assessment of their antioxidant, antimicrobial and cytotoxic activities. IET Nanobiotechnol. 2016; 10(6):438-444. DOI: https://doi.org/10.1049/iet-nbt.2015.0104

Mishra K., Ojha H. and Chaudhury N.K. Estimation of antiradical properties of antioxidants using DPPH assay: a critical review and results. Food Chem. 2012; 130(4):1036-1043. DOI: https://doi.org/10.1016/j.foodchem.2011.07.127

Kharat S.N. and Mendhulkar V.D. Synthesis, characterization and studies on antioxidant activity of silver nanoparticles using Elephantopus scaber leaf extract. Mater. Sci. Eng. C 2016; 62:719-724.‏ DOI: https://doi.org/10.1016/j.msec.2016.02.024

Divya K.S., Harshitha B.S., Kumar D. and Chauhan J.B. In vitro investigation of antioxidant potentiality of methanol and silver nanoparticles extract from Trigonella foenum-graecum. J. Pharmacogn. Phytochem. 2019; 8(3):2213-2221.

Noginov M.A., Zhu G., Bahoura M., Adegoke J., Small C., Ritzo B.A., et al. The effect of gain and absorption on surface plasmons in metal nanoparticles. Appl. Phys. B 2007; 86:455-460. DOI: https://doi.org/10.1007/s00340-006-2401-0

Nayak D., Ashe S., Rauta P.R., Kumari M. and Nayak B. Bark extract mediated green synthesis of silver nanoparticles: evaluation of antimicrobial activity and antiproliferative response against osteosarcoma. Mater. Sci. Eng. C 2016; 58:44-52. DOI: https://doi.org/10.1016/j.msec.2015.08.022

Raja S., Ramesh V. and Thivaharan V. Green biosynthesis of silver nanoparticles using Calliandra haematocephala leaf extract, their antibacterial activity and hydrogen peroxide sensing capability. Arab. J. Chem. 2017; 10(2):253-261. DOI: https://doi.org/10.1016/j.arabjc.2015.06.023

Rivera-Rangel R.D., González-Muñoz M.P., Avila-Rodriguez M., Razo-Lazcano T.A. and Solans C. Green synthesis of silver nanoparticles in oil-in-water microemulsion and nano-emulsion using geranium leaf aqueous extract as a reducing agent. Colloids Surf. A Physicochem. Eng. Asp. 2018; 536:60-67. DOI: https://doi.org/10.1016/j.colsurfa.2017.07.051

Khoshnamvand M., Huo C. and Liu J. Silver nanoparticles synthesized using Allium ampeloprasum L. leaf extract: characterization and performance in catalytic reduction of 4-nitrophenol and antioxidant activity. J. Mol. Struct. 2019; 1175:90-96. DOI: https://doi.org/10.1016/j.molstruc.2018.07.089

Lam V.P., Beomseon L., Anh V.K., Loi D.N., Kim S., Kwang-Ya L. and Park J. Effectiveness of silver nitrate application on plant growth and bioactive compounds in Agastache rugosa (Fisch. & CA Mey.) kuntze. Heliyon. 2023; 9(9). DOI: https://doi.org/10.1016/j.heliyon.2023.e20205

Kim S.T., Saha K., Kim C. and Rotello V.M. The role of surface functionality in determining nanoparticle cytotoxicity. Acc. Chem. Res. 2013; 46(3):681-691.‏ DOI: https://doi.org/10.1021/ar3000647

Zazharskyi V.V., Davydenko P., Kulishenko O., Borovik I.V., Zazharska N.M. and Brygadyrenko V.V. Antibacterial and fungicidal activities of ethanol extracts of 38 species of plants. Biosyst. Divers. 2020; 28(3):281-289. DOI: https://doi.org/10.15421/012037

Zazharskyi V.V., Davydenko P.O., Kulishenko O.M., Borovik I.V. and Brygadyrenko V.V. Antibacterial and fungicidal activities of ethanol extracts from Cotinus coggygria, Rhus typhina, R. trilobata, Toxicodendron orientale, Hedera helix, Aralia elata, Leptopus chinensis and Mahonia aquifolium. Regul. Mech. Biosyst. 2020; 11(2). DOI: https://doi.org/10.15421/022046

Reddy N.J., Vali D.N., Rani M. and Rani S.S. Evaluation of antioxidant, antibacterial and cytotoxic effects of green synthesized silver nanoparticles by Piper longum fruit. Mater. Sci. Eng. C. 2014; 34:115-122. DOI: https://doi.org/10.1016/j.msec.2013.08.039

Feng Q.L., Wu J., Chen G.Q., Cui F.Z., Kim T.N. and Kim J.O. A mechanistic study of the antibacterial effect of silver ions on Escherichia coli and Staphylococcus aureus. J. Biomed. Mater. Res. 2000; 52(4):662-668. DOI: https://doi.org/10.1002/1097-4636(20001215)52:4<662::AID-JBM10>3.0.CO;2-3

Medici S., Peana M., Nurchi V.M. and Zoroddu M.A. Medical uses of silver: history, myths, and scientific evidence. J. Med. Chem. 2019; 62(13):5923-5943. DOI: https://doi.org/10.1021/acs.jmedchem.8b01439

Zhang X.F., Liu Z.G., Shen W. and Gurunathan S. Silver nanoparticles: synthesis, characterization, properties, applications, and therapeutic approaches. Int. J. Mol. Sci. 2016; 17(9):1534. DOI: https://doi.org/10.3390/ijms17091534

Abdalla A.M., Xiao L., Ullah M.W., Yu M., Ouyang C. and Yang G. Current challenges of cancer anti-angiogenic therapy and the promise of nanotherapeutics. Theranostics. 2018; 8(2):533. DOI: https://doi.org/10.7150/thno.21674

Simon S., Sibuyi N.R.S., Fadaka A.O., Meyer S., Josephs J., Onani M.O., et al. Biomedical applications of plant extract-synthesized silver nanoparticles. Biomedicines. 2022; 10(11):2792 DOI: https://doi.org/10.3390/biomedicines10112792