GREEN SYNTHESIS OF CHITOSAN-MEDIATED SILVER NANOCOMPOSITE USING PIPER BETEL STEM EXTRACT: EVALUATING ANTIBACTERIAL AND ANTICANCER ACTIVITY
DOI:
https://doi.org/10.22159/ajpcr.2026v19i1.56502Keywords:
green nanoparticle, Nanocomposites, piper betel, Chitosan, Anticancer activity, AntibacterialAbstract
Objective: The integration of herb-based nanomedicine has gained significant attention, aiming to improve therapeutic outcomes while minimizing drug toxicity. Piper betel is a medicinal herb widely used in traditional therapy due to its potent anticancer, antimicrobial, antioxidant, and anti-inflammatory properties.
Methods: P. betel stem extract is used to synthesize chitosan-mediated silver nanocomposite (AgNC). Characterization was done using UV-visible spectrophotometry, Fourier transform infrared (FTIR), X-ray diffraction, dynamic light scattering with Zeta potential, and FESEM with energy dispersive X-ray. Its antibacterial activity was evaluated in Staphylococcus aureus, Pseudomonas aeruginosa, and Escherichia coli, as well as antioxidant diphenyl-2-picrylhydrazyl and anti-inflammatory assays (egg albumin and bovine serum albumin) were performed. Further, its anticancer activity was evaluated in A549 and Henrietta Lacks (HeLa) cell lines.
Results: AgNC exhibited a sharp surface plasmon resonance peak at 450 nm. FTIR analysis revealed various peaks. Scanning electron microscopy analysis revealed a soft-fibrous sheet-like morphology. In antibacterial activity, a significant zone of inhibition was observed against P. aeruginosa and E. coli (16 mm). In addition, AgNC exhibited inhibition of free radicals and protein denaturation. In anticancer activity, it showed greater activity against A549 (66.7±5.9 μg/mL) than HeLa (85.7±7.8 μg/mL), and reduced cell migration within 48 h at 80 μg/mL in A549 cell lines.
Conclusion: These findings highlight AgNCs as the most promising candidates for biomedical applications, particularly for antibacterial and anticancer therapy. However, further investigations are required to elucidate the precise molecular mechanisms of therapeutic efficacies and modifications to enhance their anticancer activity.
Downloads
References
1. Rai M, Pandit R, Gaikwad S, Yadav A, Gade A. Potential applications of curcumin and curcumin nanoparticles: From traditional therapeutics to modern nanomedicine. Nanotechnol Rev. 2015;4(2):161-72. doi: 10.1515/ntrev-2015-0001
2. Rajeshkumar S, Bharath LV, Geetha R. Broad spectrum antibacterial silver nanoparticle green synthesis: Characterization, and mechanism of action. In: Green Synthesis, Characterization and Applications of Nanoparticles. Netherlands: Elsevier; 2019. p. 429-44. doi: 10.1016/ B978-0-08-102579-6.00018-6
3. Goswami MJ, Hati Boruah JL, Saikia R, Dutta U, Kakati D. Synthesis, antimicrobial and antioxidant bioevaluation of silver nanoparticles using leaf extract of Sarcochlamys pulcherrima. Next Nanotechnol. 2024;5:100063. doi: 10.1016/j.nxnano.2024.100063
4. Kanniah P, Chelliah P, Thangapandi JR, Gnanadhas G, Mahendran V, Robert M. Green synthesis of antibacterial and cytotoxic silver nanoparticles by Piper nigrum seed extract and development of antibacterial silver-based chitosan nanocomposite. Int J Biol Macromol. 2021;189:18-33. doi: 10.1016/j.ijbiomac.2021.08.056, PMID 34389391
5. El-Meligy MA, Abd El-Monaem EM, Eltaweil AS, Mohy-Eldin MS, Ziora ZM, Heydari A, et al. Recent advancements in metallic Au- and Ag-based chitosan nanocomposite derivatives for enhanced anticancer drug delivery. Molecules. 2024;29(10):2393. doi: 10.3390/ molecules29102393, PMID 38792255
6. Singh T, Singh P, Pandey VK, Singh R, Dar AH. A literature review on bioactive properties of betel leaf (Piper betel L.) and its applications in food industry. Food Chem Adv. 2023;3:100536. doi: 10.1016/j. focha.2023.100536
7. Sonphakdi T, Tani A, Payaka A, Ungcharoenwiwat P. Antibacterial and toxicity studies of phytochemicals from Piper betle leaf extract. J King Saud Univ Sci. 2024;36(10):103430. doi: 10.1016/j.jksus.2024.103430
8. Alven S, Aderibigbe BA. Chitosan-based scaffolds incorporated with silver nanoparticles for the treatment of infected wounds. Pharmaceutics. 2024;16(3):327. doi: 10.3390/pharmaceutics16030327, PMID 38543221
9. Sivanesan I, Gopal J, Muthu M, Shin J, Mari S, Oh J. Green synthesized chitosan/chitosan nanoforms/nanocomposites for drug delivery applications. Polymers (Basel). 2021;13(14):2256. doi: 10.3390/ polym13142256, PMID 34301013
10. Biswas P, Anand U, Saha SC, Kant N, Mishra T, Masih H, et al. Betelvine (Piper betle L.): A comprehensive insight into its ethnopharmacology, phytochemistry, and pharmacological, biomedical and therapeutic attributes. J Cell Mol Med. 2022;26(11):3083-119. doi: 10.1111/jcmm.17323, PMID 35502487
11. Kalaivani R, Maruthupandy M, Muneeswaran T, Hameedha Beevi A, Anand M, Ramakritinan CM, et al. Synthesis of chitosan-mediated silver nanoparticles (Ag NPs) for potential antimicrobial applications. Front Lab Med. 2018;2(1):30-5. doi: 10.1016/j.flm.2018.04.002
12. Badawy ME, Lotfy TM, Shawir SM. Preparation and antibacterial activity of chitosan-silver nanoparticles for application in preservation of minced meat. Bull Natl Res Cent. 2019;43(1):83. doi: 10.1186/ s42269-019-0124-8
13. Nguyen TT, Le TQ, Nguyen TT, Nguyen LT, Nguyen DT, Tran TV. Characterizations and antibacterial activities of passion fruit peel pectin/chitosan composite films incorporated Piper betle L. leaf extract for preservation of purple eggplants. Heliyon. 2022;8(8):e10096. doi: 10.1016/j.heliyon.2022.e10096, PMID 36016528
14. Tamboli M, Tamboli N. Exploration of in-vitro anti-cancer activity of Piper betel Linn. SSRN J. 2024;11(2):157-165. doi: 10.2139/ ssrn.4854818
15. Veettil SR, Sunil EA, Mukunda A, Mohan A, John S, Pynadath MK. Anticancer effect of Piper betle leaf extract on KB cell lines - an in vitro study. Oral Maxillofac Pathol J. 2022;13(1):28-31.
16. Mohamad N, Rahman A, Sheikh Abdul Kadir SH. Hydroxychavicol as a potential anticancer agent [review]. Oncol Lett. 2022;5;25(1):34. doi: 10.3892/ol.2022.13620
17. Yoonus J, Resmi R, Beena B. Greener nanoscience: Piper betel leaf extract mediated synthesis of CaO nanoparticles and evaluation of its antibacterial and anticancer activity. Mater Today Proc. 2021;41:535-40. doi: 10.1016/j.matpr.2020.05.246
18. Tagrida M, Nilsuwan K, Gulzar S, Prodpran T, Benjakul S. Fish gelatin/chitosan blend films incorporated with betel (Piper betle L.) leaf ethanolic extracts: Characteristics, antioxidant and antimicrobial properties. Food Hydrocoll. 2023;137:108316. doi: 10.1016/j. foodhyd.2022.108316
19. El-Naggar NE, Shiha AM, Mahrous H, Mohammed AB. Green synthesis of chitosan nanoparticles, optimization, characterization and antibacterial efficacy against multi drug resistant biofilm-forming Acinetobacter baumannii. Sci Rep. 2022;12(1):19869. doi: 10.1038/ s41598-022-24303-5, PMID 36400832
20. Charirak P, Prajantasan R, Premprayoon K, Srikacha N, Ratananikom K. In vitro antibacterial activity and mode of action of Piper betle extracts against soft rot disease-causing bacteria. Scientifica. 2023;2023(1):5806841. doi: 10.1155/2023/5806841, PMID 37766936
21. Sumithra M, Aparna Y, Raghavendra Rao P. Morphological growth study of extended branches of silver nanodendrites using Piper betel leaf extract. Mater Today Proc. 2023;92:671-8. doi: 10.1016/j. matpr.2023.04.176
22. Li CX, Elhassan GO, Abdoun SA, Khan RA, Goyal M, Bansal M, et al. Ultra-fast synthesis of curcumin-loaded silver nanoparticles: Improved physicochemical properties for drug delivery. Int J App Pharm. 2025;17(1):216-23. doi: 10.22159/ijap.2025v17i1.52647
23. Mirda E, Idroes R, Khairan K, Tallei TE, Ramli M, Earlia N, et al. Synthesis of chitosan-silver nanoparticle composite spheres and their antimicrobial activities. Polymers (Basel). 2021;13(22):3990. doi: 10.3390/polym13223990, PMID 34833288
24. Arif D, Niazi MB, Ul-Haq N, Anwar MN, Hashmi E. Preparation of antibacterial cotton fabric using chitosan-silver nanoparticles. Fibers Polym. 2015;16(7):1519-26. doi: 10.1007/s12221-015-5245-6
25. Malarkodi C, Rajeshkumar S, Annadurai G. Detection of environmentally hazardous pesticide in fruit and vegetable samples using gold nanoparticles. Food Control. 2017;80:11-8. doi: 10.1016/j. foodcont.2017.04.023
26. Balouiri M, Sadiki M, Ibnsouda SK. Methods for in vitro evaluating antimicrobial activity: A review. J Pharm Anal. 2016;6(2):71-9. doi: 10.1016/j.jpha.2015.11.005, PMID 29403965
27. Kakian F, Mirzaei E, Moattari A, Takallu S, Bazargani A. Determining the cytotoxicity of the minimum inhibitory concentration (MIC) of silver and zinc oxide nanoparticles in ESBL and carbapenemase producing Proteus mirabilis isolated from clinical samples in Shiraz, Southwest Iran. BMC Res Notes. 2024;17(1):40. doi: 10.1186/s13104- 023-06402-2, PMID 38287416
28. Topçu S, Şeker MG. In vitro antimicrobial effects and inactivation mechanisms of 5,8-dihydroxy-1,4-napthoquinone. Antibiotics (Basel). 2022;11(11):1537. doi: 10.3390/antibiotics11111537, PMID 36358192
29. Munusamy T, Shanmugam R. Green synthesis of copper oxide nanoparticles synthesized by Terminalia chebula dried fruit extract: Characterization and antibacterial action. Cureus. 2023;15(12):e50142. doi: 10.7759/cureus.50142, PMID 38186403
30. Dhanraj G, Rajeshkumar S. Anticariogenic effect of selenium nanoparticles synthesized using Brassica oleracea. J Nanomater. 2021;2021(1):1-9. doi: 10.1155/2021/8115585
31. Chandra S, Chatterjee P, Dey P, Bhattacharya S. Evaluation of in vitro anti-inflammatory activity of coffee against the denaturation of protein. Asian Pac J Trop Biomed. 2012;2(1):S178-80. doi: 10.1016/S2221- 1691(12)60154-3
32. Ameena M, Arumugham IM, Ramalingam K, Rajeshkumar S. Evaluation of the anti-inflammatory, antimicrobial, antioxidant, and cytotoxic effects of chitosan thiocolchicoside-lauric acid nanogel. Cureus. 2023;15(9):e46003. doi: 10.7759/cureus.46003, PMID 37900405
33. Swidan NS, Hashem YA, Elkhatib WF, Yassien MA. Antibiofilm activity of green synthesized silver nanoparticles against biofilmassociated enterococcal urinary pathogens. Sci Rep. 2022;12(1):3869. doi: 10.1038/s41598-022-07831-y, PMID 35264654
34. Available from: https://manuals/man0019028_cyquant_mtt_ cellproliferationassaykit_pi.pdf
35. Nandhini JT, Ezhilarasan D, Rajeshkumar S. An ecofriendly synthesized gold nanoparticles induces cytotoxicity via apoptosis in HEPG2 cells. Environ Toxicol. 2021;36(1):24-32. doi: 10.1002/tox.23007, PMID 32794643
36. Grada A, Otero-Vinas M, Prieto-Castrillo F, Obagi Z, Falanga V. Research techniques made simple: Analysis of collective cell migration using the wound healing assay. J Invest Dermatol. 2017;137(2):e11-6. doi: 10.1016/j.jid.2016.11.020, PMID 28110712
37. Ermawati DE, Yugatama A, Ramadhani BR, Pertiwi I, Rosikhoh A, Novachiria SR. Stability and antibacterial activity test of nanosilver biosynthetic hydrogel. Int J App Pharm. 2022;14(2):221-6. doi: 10.22159/ijap.2022v14i2.43584
38. Amar AA, Putra ED, Satria D, Sitorus P, Lee HL, Waruwu SB, et al. Biosynthesis and biological activity of silver nanoparticles from Zanthoxylum acanthopodium fruits. S Afr J Chem Eng. 2025;52:271-81. doi: 10.1016/j.sajce.2025.03.002
39. Wang LS, Wang CY, Yang CH, Hsieh CL, Chen SY, Shen CY, et al. Synthesis and anti-fungal effect of silver nanoparticles-chitosan composite particles. Int J Nanomedicine. 2015;10:2685-96. doi: 10.2147/IJN.S77410, PMID 25878501
40. Govindan S, Nivethaa EA, Saravanan R, Narayanan V, Stephen A. Synthesis and characterization of chitosan-silver nanocomposite. Appl Nanosci. 2012;2(3):299-303. doi: 10.1007/s13204-012-0109-5
41. Kulikouskaya V, Hileuskaya K, Kraskouski A, Kozerozhets I, Stepanova E, Kuzminski I, et al. Chitosan-capped silver nanoparticles: A comprehensive study of polymer molecular weight effect on the reaction kinetic, physicochemical properties, and synergetic antibacterial potential. SPE Polym. 2022;3(2):77-90. doi: 10.1002/ pls2.10069
42. Saadh MJ. Silver nanoparticles inhibit infectious bronchitis virus replication. Int J App Pharm. 2023;15(6):163-6. doi: 10.22159/ ijap.2023v15i6.48963
43. Mannopantar SR, Bendigeri HH, Kulkarni VK, Patil VS, Manjunatha DH, Kalasad MN. Preparation of colloidal Ag nanoparticles. Mater Today Proc. 2022;60:1156-9. doi: 10.1016/j.matpr.2022.03.152
44. Jalab J, Abdelwahed W, Kitaz A, Al-Kayali R. Green synthesis of silver nanoparticles using aqueous extract of Acacia cyanophylla and its antibacterial activity. Heliyon. 2021;7(9):e08033. doi: 10.1016/j. heliyon.2021.e08033, PMID 34611564
45. Ogungbesan SO, Buxaderas E, Adedokun RA, Moglie Y, Grijalvo S, García MT, et al. Synthesis and characterization of chitosan-silver nanocomposite film: antibacterial and cytotoxicity study. ChemistrySelect. 2024;9(42):e202404909. doi: 10.1002/slct.202404909
46. Sherif HH, Khalil SK, Hegazi AG, Khalil WA, Moharram MA. Factors affecting the antibacterial activity of chitosan-silver nanocomposite. IET Nanobiotechnol. 2017;11(6):731-7. doi: 10.1049/iet-nbt.2016.0249
47. Vas NV, Jain RK, Kaliaperumal K. An in-vitro analysis of the mechanical and anti-bacterial properties of betel leaf extract with chitosan coating on orthodontic aligners. Pesqui Bras Odontopediatr Clin Integr. 2024 Nov;25:e230243.
48. dos Santos EM, Martins CC, de Oliveira Santos JV, da Silva WR, Silva SB, Pelagio-Flores MA, et al. Silver nanoparticles-chitosan composites activity against resistant bacteria: Tolerance and biofilm inhibition. J Nanopart Res. 2021;23(8):196. doi: 10.1007/s11051-021- 05314-1, PMID 34456615
49. Xu J, Li Y, Wang H, Zhu M, Feng W, Liang G. Enhanced antibacterial and anti-biofilm activities of antimicrobial peptides modified silver nanoparticles. Int J Nanomedicine. 2021;16:4831-46. doi: 10.2147/IJN. S315839, PMID 34295158
50. Bruna T, Maldonado-Bravo F, Jara P, Caro N. Silver nanoparticles and their antibacterial applications. Int J Mol Sci. 2021;22(13):7202. doi: 10.3390/ijms22137202, PMID 34281254
51. Muneeswaran T, Maruthupandy M, Mary AS, Vennila T, Rajaram K, Ramakritinan CM, et al. Starch-mediated synthesis of chitosan/silver nanocomposites for antibacterial, antibiofilm and wound healing applications. J Drug Deliv Sci Technol. 2023;84:104424. doi: 10.1016/j. jddst.2023.104424
52. Hussein HS, Ngugi C, Tolo FM, Maina EN. Anticancer potential of silver nanoparticles biosynthesized using Catharanthus roseus leaves extract on cervical (HeLa229) cancer cell line. Sci Afr. 2024;25:e02268. doi: 10.1016/j.sciaf.2024.e02268
53. Xu G, Yu H, Shi X, Sun L, Zhou Q, Zheng D, et al. Cisplatin sensitivity is enhanced in non-small cell lung cancer cells by regulating epithelial-mesenchymal transition through inhibition of eukaryotic translation initiation factor 5A2. BMC Pulm Med. 2014;14(1):174. doi: 10.1186/1471-2466-14-174, PMID 25380840
54. El-Sheikh SM, Edrees N, EL-Sayed H, Khamis T, Arisha AH, Metwally MM, et al. Could cisplatin loading on biosynthesized silver nanoparticles improve its therapeutic efficacy on human prostate cancer cell line and reduce its in vivo nephrotoxic effects? Biol Trace Elem Res. 2022;200(2):582-90. doi: 10.1007/s12011-021-02677-3, PMID 33759109
55. Gopinath V, MubarakAli D, Vadivelu J, Manjunath Kamath S, Syed A, Elgorban AM. Synthesis of biocompatible chitosan-decorated silver nanoparticles biocomposites for enhanced antimicrobial and anticancer property. Process Biochem. 2020;99:348-56. doi: 10.1016/j. procbio.2020.09.011
56. Preethi R, Padma PR. Anticancer activity of silver nanobioconjugates synthesised from Piper betle leaves extract and its active compound eugenol. Int J Pharm Pharm Sci. 2016;8(9):201. doi: 10.22159/ ijpps.2016.v8i9.12993
57. Milliana A, Sari RA, Miranda SA, Mutiah R. Evaluation of anticancer activity and mechanism of action of myricetin on HeLa, T47D, and Vero cells: Comparative analysis with cisplatin and doxorubicin. Biomed Pharmacol J. 2025;18(1):835-47. doi: 10.13005/bpj/3133
58. Zhang XF, Shen W, Gurunathan S. Silver nanoparticle-mediated cellular responses in various cell lines: An in vitro model. Int J Mol Sci. 2016;17(10):1603. doi: 10.3390/ijms17101603, PMID 27669221
Published
How to Cite
Issue
Section
Copyright (c) 2025 Naresh Kumar kodiganti, Nalini Devarajan, Pavithra Amritkumar, Tharani Munusamy, Malchi Suresh, Manju Bargavi S
This work is licensed under a Creative Commons Attribution 4.0 International License.
The publication is licensed under CC By and is open access. Copyright is with author and allowed to retain publishing rights without restrictions.