The Journal of Phytopharmacology 2025; 14(4):274-280 DOI:10.31254/phyto.2025.14407

Research Article

Cytotoxic and apoptotic potential of biosynthesised silver nanoparticles in Dalton’s lymphoma ascites cells

Preethy John¹ , Nisha A. R.² , Suresh N Nair³ , Bibu John Kariyil⁴ , Uma Radhakrishnan⁵

Preethy John,Nisha A. R.,Suresh N Nair,Bibu John Kariyil,Uma Radhakrishnan

1. Associate Professor, Department of Veterinary Pharmacology and Toxicology, College of Veterinary and Animal Sciences, Mannuthy- 680651, Kerala, India
2. Professor and Head, Department of Veterinary Pharmacology and Toxicology, College of Veterinary and Animal Sciences, Pookode- 673576, Kerala, India
3. Associate Professor, Department of Veterinary Pharmacology and Toxicology, College of Veterinary and Animal Sciences, Mannuthy- 680651, Kerala, India
4. Assistant Professor, Department of Veterinary Pharmacology and Toxicology, College of Veterinary and Animal Sciences, Mannuthy- 680651, Kerala, India
5. Head, Department of Veterinary Biochemistry and Molecular Biology, College of Veterinary and animal Sciences, Mannuthy- 680651, Kerala, India

*Author to whom correspondence should be addressed.

Received: 9th May, 2025 / Accepted: 27th August, 2025 / Published : 30th September, 2025

Abstract

Background: Nanoparticle-mediated cancer therapies have gained increasing attention due to their targeted action and reduced side effects. Piperlongumine (PPLM), a natural alkaloid, has shown anticancer potential, and its combination with nanotechnology could enhance therapeutic outcomes. Dalton’s Lymphoma Ascites (DLA) cells serve as a reliable model for evaluating antineoplastic agents. Objective: To evaluate and compare the cytotoxic and apoptotic potential of biosynthesised silver nanoparticles (P-AgNPs), chemically synthesised silver nanoparticles (C-AgNPs), piperlongumine (PPLM), and the standard chemotherapeutic agent 5-fluorouracil (5-FU) against DLA cells, and to elucidate the probable underlying mechanisms of cell death. Materials and Methods: The cytotoxicity of test compounds was assessed using the MTT assay, and IC?? values were calculated. Trypan blue dye exclusion assay was performed to evaluate cell viability after 3 hours of exposure. Apoptotic changes were observed through Acridine Orange/Ethidium Bromide (AO/EB) dual staining after 24-hour treatment with IC?? concentrations. Mechanistic studies included DCF-DA assay for intracellular ROS levels, JC-1 staining for mitochondrial membrane potential, DNA fragmentation assay for DNA damage, and qRT-PCR for gene expression analysis of Caspase-3, p53, and Bcl-2, using GAPDH as reference. Results: P-AgNPs showed the highest cytotoxicity with an IC?? of 1.25 µg/mL, outperforming PPLM (4.028 µg/mL), 5-FU (326.20 µg/mL), and C-AgNPs (743.60 µg/mL). Trypan blue assay indicated an average reduction in cell viability to 49.07% across all treatments. AO/EB staining revealed late apoptosis in P-AgNP- and PPLM-treated cells, early apoptosis with 5-FU, and minimal apoptotic effect from C-AgNPs. Mechanistic assays showed that P-AgNPs induced a marked increase in ROS and significant mitochondrial membrane depolarisation. DNA fragmentation confirmed apoptosis. Gene expression analysis showed upregulation of Caspase-3 and p53, and downregulation of Bcl-2 in P-AgNP-treated cells. Conclusion: Biosynthesised silver nanoparticles (P-AgNPs) demonstrated superior cytotoxic and apoptotic effects in DLA cells compared to chemically synthesised nanoparticles and conventional agents. The enhanced efficacy is attributed to ROS generation, mitochondrial dysfunction, and regulation of apoptotic genes, highlighting P-AgNPs as a promising candidate for anticancer therapy.