Green synthesis of silver nanoparticles using Viola odorata and Onosma hispidum: Evaluation of their antimicrobial, antioxidant, and hemolytic properties against multidrug-resistant microbes

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Green synthesis of silver nanoparticles using Viola odorata and Onosma hispidum: Evaluation of their antimicrobial, antioxidant, and hemolytic properties against multidrug-resistant microbes
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Keywords

Antimicrobial
Antimicrobial resistance
Antioxidant
Green synthesis
Hemolysis assay
Hemolytic properties
Multidrug resistant bacteria
Nanoparticles
Onosma hispidum
Silver nanoparticles
Viola odorata

How to Cite

1.
Andleeb A, Khalid A, Khalil S, Arshad H, Sadaf S. Green synthesis of silver nanoparticles using Viola odorata and Onosma hispidum: Evaluation of their antimicrobial, antioxidant, and hemolytic properties against multidrug-resistant microbes. Electron. J. Biotechnol. [Internet]. 2025 Mar. 15 [cited 2026 Jan. 26];74. Available from: https://www.ejbiotechnology.info/index.php/ejbiotechnology/article/view/2423

Abstract

Background: Rapid increase in antimicrobial resistance poses a significant threat to global health, necessitating alternative strategies to combat multi-drug resistant (MDR) pathogens. Advances in nanobiotechnology may offer promising solutions to this pressing issue. This study revolves around developing an efficient method for synthesizing silver nanoparticles (AgNPs) based on aqueous extracts of Viola odorata leaves (VoL) and Onosma hispidum root barks (OhRb), traditional medicinal perennial herbs, followed by evaluating their antioxidant and antimicrobial potential against MDR pathogens.

Results: The synthesis of nanoparticles was completed in 15–20 min at 55–60°C using green approach. The characterized AgNPs showed significant antimicrobial activity against Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, Staphylococcus aureus, Streptococcus agalactiae, and Candida albicans, with zones of inhibition ranging from 12 mm ± 0.636 to 25 mm ± 0.282. Minimum inhibitory concentration and minimum bactericidal concentrations ranged from 0.39 to 6.125 µg/mL. Additionally, the nanoparticles demonstrated significant antioxidant potential compared to the aqueous plant extracts. Importantly, the investigation into the effects of the nanoparticles on red blood cells revealed no pronounced hemolysis even at higher concentrations (300 µg/mL) of VoL-AgNPs and OhRb-AgNPs, demonstrating their biocompatibility and hemolytic safety profile.

Conclusions: The VoL-AgNPs and OhRb-AgNPs exhibit appreciable antimicrobial and antioxidant activities with minimal hemolysis, underscoring their potential to combat MDR pathogens, and further applicability in therapeutic settings. However, in vivo studies are warranted to explore their biomedical applications, such as in wound dressings and treatments for disorders caused by free radical damage.

https://doi.org/10.1016/j.ejbt.2024.12.003
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References

Ajulo S, Awosile B. Global antimicrobial resistance and use surveillance system (GLASS 2022): Investigating the relationship between antimicrobial resistance and antimicrobial consumption data across the participating countries. PLoS ONE 2024;19(2):e0297921. https://doi.org/10.1371/journal.pone.0297921 PMid: 38315668

Oliveira M, Antunes W, Mota S, et al. An overview of the recent advances in antimicrobial resistance. Microorganisms 2024;12(9):1920. https://doi.org/10.3390/microorganisms12091920 PMid: 39338594

Serweci?ska L. Antimicrobials and antibiotic-resistant bacteria: A risk to the environment and to public health. Water 2020;12(12):3313. https://doi.org/10.3390/w12123313

Bayda S, Adeel M, Tuccinardi T, et al. The history of nanoscience and nanotechnology: From chemical-physical applications to nanomedicine. Molecules 2019;25(1):112. https://doi.org/10.3390/molecules25010112 PMid: 31892180

Natan M, Banin E. From nano to micro: Using nanotechnology to combat microorganisms and their multidrug resistance. FEMS Microbiol Rev 2017;41(3):302-322. https://doi.org/10.1093/femsre/fux003 PMid: 28419240

Aguilar-Garay R, Lara-Ortiz LF, Campos-López M, et al. A comprehensive review of silver and gold nanoparticles as effective antibacterial agents. Pharmaceuticals 2024;17(9):1134. https://doi.org/10.3390/ph17091134 PMid: 39338299

Arshad H, Saleem M, Pasha U, et al. Synthesis of Aloe vera-conjugated silver nanoparticles for use against multidrug-resistant microorganisms. Electron J Biotechnol 2022;55:55-64. https://doi.org/10.1016/j.ejbt.2021.11.003

Altammar KA. A review on nanoparticles: Characteristics, synthesis, applications, and challenges. Front Microbiol 2023;14:1155622. https://doi.org/10.3389/fmicb.2023.1155622 PMid: 37180257

Naysmith A, Mian NS, Rana S. Development of conductive textile fabric using Plackett–Burman optimized green synthesized silver nanoparticles and in situ polymerized polypyrrole. Green Chem Lett Rev 2023;16(1):2158690. https://doi.org/10.1080/17518253.2022.2158690

Laib I, Ali BD, Boudebia O. Green synthesis of silver nanoparticles using Helianthemum lippii extracts (Hl-NPs): Characterization, antioxidant and antibacterial activities, and study of interaction with DNA. Journal of Organometallic Chemistry 2023;986:122619. https://doi.org/10.1016/j.jorganchem.2023.122619

Samreen, Ahmad I, Khan SA, et al. Green synthesized silver nanoparticles from Phoenix dactylifera synergistically interact with bioactive extract of Punica granatum against bacterial virulence and biofilm development. Microbial Pathogenesis 2024;192:106708. DOI: https://doi.org/10.1016/j.micpath.2024.106708 PMid: 38782213.

Abbas M, Hussain T, Iqbal J, et al. Synthesis of silver nanoparticle from Allium sativum as an eco-benign agent for biological applications. Polish Journal of Environmental Studies 2022;31(1):533-538. https://doi.org/10.15244/pjoes/135764

Zhao H, Su H, Ahmeda A, et al. Biosynthesis of copper nanoparticles using Allium eriophyllum Boiss leaf aqueous extract: Characterization and analysis of their antimicrobial and cutaneous wound-healing potentials. Applied Organometallic Chemistry 2022;36(12);e5587. https://doi.org/10.1002/aoc.5587

Tavanappanavar AN, Mulla SI, Seth SC, et al. Phytochemical analysis, GC-MS profile and determination of antibacterial, antifungal, anti-inflammatory, antioxidant activities of peel and seeds extracts (chloroform and ethyl acetate) of Tamarindus indica L. Saudi Journal of Biological Sciences 2024;31(1):103878. https://doi.org/10.1016/j.sjbs.2023.103878 PMid: 38125735

Dehnoee A, Kalbasi JR, Zangeneh MM, et al. Characterization, anti-lung cancer activity, and cytotoxicity of bio-synthesized copper nanoparticles by Thymus fedtschenkoi leaf extract. Journal of Cluster Science 2024;35:863–874. https://doi.org/10.1007/s10876-023-02512-w

Wang G, Ahmeda A, Malek Z, et al. Chemical characterization and therapeutic properties of Achillea biebersteinii leaf aqueous extract synthesized copper nanoparticles against methamphetamine-induced cell death in PC12: A study in the nanotechnology and neurology fields. Applied Organometallic Chemistry 2020;34(4):e5488. https://doi.org/10.1002/aoc.5488

Alshameri AW. Owais M, Altaf I, et al. Rumex nervosus mediated green synthesis of silver nanoparticles and evaluation of its in vitro antibacterial, and cytotoxic activity. OpenNano 2022;8:100084. https://doi.org/10.1016/j.onano.2022.100084

Khane Y, Benouis K, Albukhaty S, et al. Green synthesis of silver nanoparticles using aqueous Citrus limon zest extract: Characterization and evaluation of their antioxidant and antimicrobial properties. Nanomaterials 2022;12(12):2013. https://doi.org/10.3390/nano12122013 PMid: 35745352

Jalab J, Abdelwahed W, Kitaz A, et al. Green synthesis of silver nanoparticles using aqueous extract of Acacia cyanophylla and its antibacterial activity. Heliyon 2021;7(9):e08033. https://doi.org/10.1016/j.heliyon.2021.e08033 PMid: 34611564

Hashemi Z, Mohammadyan M, Naderi S, et al. Green synthesis of silver nanoparticles using Ferula persica extract (Fp-NPs): Characterization, antibacterial, antileishmanial, and in vitro anticancer activities. Material Today Communications 2021;27:102264. https://doi.org/10.1016/j.mtcomm.2021.102264

Dehnoee A, Kalbasi RJ, Zangeneh MM, et al. One-step synthesis of silver nanostructures using Heracleum persicum fruit extract, their cytotoxic activity, anti-cancer and anti-oxidant activities. Micro & Nano Letters 2023;18(1):e12153. https://doi.org/10.1049/mna2.12153

Salwan R, Rana A, Saini R, et al. Diversity analysis of endophytes with antimicrobial and antioxidant potential from Viola odorata: An endemic plant species of the Himalayas. Brazilian Journal of Microbiology 2023;54(3):2361-2374. https://doi.org/10.1007/s42770-023-01010-5 PMid: 37227628

Zahra T, Ahmad KS, Sharif S. Identification and implication of organic compounds of Viola odorata: A potential source for bio-fabrication of nickel oxide nanoparticles. Applied Nanoscience 2021;11:1593-1603. https://doi.org/10.1007/s13204-021-01777-9

Kumar N, Kumar R, Kishore K. Onosma L.: A review of phytochemistry and ethnopharmacology. Pharmacognosy Review 2013;7(14):140-151. https://doi.org/10.4103/0973-7847.120513 PMid: 24347922

Wazir NU, Khan AI, Javed A, et al. Onosma hispidum L. extract reverses hyperlipidemia, hypertension, and associated vascular dysfunction in rats. Saudi Journal of Biological Sciences 2023;30(8):103712. https://doi.org/10.1016/j.sjbs.2023.103712 PMid: 37405138

Arshad H, Sami MA, Sadaf S, et al. Salvadora persica mediated synthesis of silver nanoparticles and their antimicrobial efficacy. Scientific Reports 2021;11:5996. https://doi.org/10.1038/s41598-021-85584-w

Arshad H, Sadaf S, Hassan U. De-novo fabrication of sunlight irradiated silver nanoparticles and their efficacy against E. coli and S. epidermidis. Scientific Reports 2022;12:676. https://doi.org/10.1038/s41598-021-04674-x PMid: 35027620

Khare S, Singh RK, Prakash O. Green synthesis, characterization and biocompatibility evaluation of silver nanoparticles using radish seeds. Results in Chemistry 2022;4:100447. https://doi.org/10.1016/j.rechem.2022.100447

Sreelekha E, George B, Shyam A, et al. A comparative study on the synthesis, characterization, and antioxidant activity of green and chemically synthesized silver nanoparticles. BioNanoSci 2021;11:489-496. https://doi.org/10.1007/s12668-021-00824-7

Parthiban E, Manivannan N, Ramanibai R, et al. Green synthesis of silver-nanoparticles from Annona reticulata leaves aqueous extract and its mosquito larvicidal and anti-microbial activity on human pathogens. Biotechnology Reports 2018;21:e00297. https://doi.org/10.1016/j.btre.2018.e00297 PMid: 30581768

Karnjana K, Jewboonchu J, Niyomtham N, et al. The potency of herbal extracts and its green synthesized nanoparticle formulation as antibacterial agents against Streptococcus mutans associated biofilms. Biotechnology Reports 2023;37:e00777. https://doi.org/10.1016/j.btre.2022.e00777 PMid: 36582762

Arshad H, Sadaf S, Hassan U. Synthesis and immobilization of silver nanoparticles on filter paper and surgical masks for antimicrobial applications. In: IEEE 17th International Conference on Nano/Micro Engineered and Molecular Systems (NEMS); 2022 April 14-17; Taoyuan, Taiwan: IEEE; 2022. p. 301-305. https://doi.org/10.1109/NEMS54180.2022.9791147

Gajendran B, Durai P, Varier KM, et al. Green synthesis of silver nanoparticle from Datura inoxia flower extract and its cytotoxic activity. BioNanoScience 2019;9:564–572. https://doi.org/10.1007/s12668-019-00645-9

George IE, Cherian T, Ragavendran C, et al. One-pot green synthesis of silver nanoparticles using brittle star Ophiocoma scolopendrina: Assessing biological potentialities of antibacterial, antioxidant, anti-diabetic and catalytic degradation of organic dyes. Heliyon 2023;9(3);e14538. https://doi.org/10.1016/j.heliyon.2023.e14538 PMid: 36967974

Clarance P, Luvankar B, Sales J, et al. Green synthesis and characterization of gold nanoparticles using endophytic fungi Fusarium solani and its in-vitro anticancer and biomedical applications. Saudi Journal of Biological Sciences 2020;27(2):706-712. https://doi.org/10.1016/j.sjbs.2019.12.026

Manosalva N, Tortella G, Diez MC, et al. Green synthesis of silver nanoparticles: Effect of synthesis reaction parameters on antimicrobial activity. World Journal of Microbiology and Biotechnology 2019;35(6):88. https://doi.org/10.1007/s11274-019-2664-3 PMid: 31134435

Vera-Nuñez LDC, Cornejo-Ruiz JO, Arenas-Chávez CA, et al. Green synthesis of a novel silver nanoparticle conjugated with Thelypteris glandulosolanosa (Raqui-Raqui): Preliminary characterization and anticancer activity. Processes 2022;10(7):1308. https://doi.org/10.3390/pr10071308

Audtarat S, Hongsachart P, Dasri T, et al. Green synthesis of silver nanoparticles loaded into bacterial cellulose for antimicrobial application. Nanocomposites 2022;8(1):34-46. https://doi.org/10.1080/20550324.2022.2055375

Erjaee H, Rajaian H, Nazifi S. Synthesis and characterization of novel silver nanoparticles using Chamaemelum nobile extract for antibacterial application. Advances in Natural Sciences: Nanoscience and Nanotechnology 2017;8(2):025004. https://doi.org/10.1088/2043-6254/aa690b

Agarwal H, Menon S, Kumar SV, et al. Mechanistic study on antibacterial action of zinc oxide nanoparticles synthesized using green route. Chemico-Biological Interactions 2018;286:60-70. https://doi.org/10.1016/j.cbi.2018.03.008 PMid: 29551637

Hassanvand A, Saadatmand S, Yazdi HL, et al. Investigation of antioxidant, antimicrobial and anticancer potential of silver nanoparticles synthesized by Viola tricolor L. extract. Journal of Agricultural Science and Technology 2022;24(4):885-900.

Santhosh A, Sandeep S, Manukumar H, et al. Green synthesis of silver nanoparticles using cow urine: Antimicrobial and blood biocompatibility studies. JCIS Open 2021;3:100023. https://doi.org/10.1016/j.jciso.2021.100023

Rashid MI, Mujawar LH, Rehan ZA et al. One-step synthesis of silver nanoparticles using Phoenix dactylifera leaves extract and their enhanced bactericidal activity. Journal of Molecular Liquids 2016;223:1114-1122. https://doi.org/10.1016/j.molliq.2016.09.030

Andleeb A, Andleeb A, Asghar S, et al. A systematic review of biosynthesized metallic nanoparticles as a promising anti-cancer-strategy. Cancers 2021;13(11):2818. https://doi.org/10.3390/cancers13112818 PMid: 34198769

Dhiman S, Singla S, Kumar I, et al. Protection of Viola odorata L. against neurodegenerative diseases: potential of the extract and major phytoconstituents. Clinical Complementary Medicine and Pharmacology 2023;3(3):100105. https://doi.org/10.1016/j.ccmp.2023.100105

Rezaei F, Tajik H, Shahbazi Y. Intelligent double-layer polymers based on carboxymethyl cellulose-cellulose nanocrystals film and poly(lactic acid)-Viola odorata petal anthocyanins nanofibers to monitor food freshness. International Journal of Biological Macromolecules 2023;252:126512. https://doi.org/10.1016/j.ijbiomac.2023.126512 PMid: 37633548

Tiwana G, Cock IE, Cheesman MJ. Phyllanthus niruri Linn.: Antibacterial activity, phytochemistry, and enhanced antibiotic combinatorial strategies. Antibiotics 2024;13(7):654. https://doi.org/10.3390/antibiotics13070654 PMid: 39061336

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