THIOCOLCHICOSIDE-ASSISTED SILVER NANOPARTICLES FOR CONTROLLING WOUND PATHOGENS: IN VITRO ANTIBIOFILM ACTIVITY, PROTEIN AND DNA LEAKAGE AGAINST PSEUDOMONAS AERUGINOSA AND ENTEROCOCCUS FAECALIS
DOI:
https://doi.org/10.22159/ajpcr.2026v19i4.58046Keywords:
Antibiofilm activity, Protein and DNA leakage, Pseudomonas aeruginosa, Thiocolchicoside-assisted AgNPs, Wound pathogensAbstract
Objective: To synthesize silver nanoparticles (AgNPs) using Gloriosa superba extract and to functionalize with thiocolchicoside, and to evaluate their antibacterial, antibiofilm, protein leakage, cytoplasmic leakage, and time-dependent antimicrobial activities against Gram-positive and Gram-negative bacteria.
Methods: Aqueous extract of G. superba was mixed with silver nitrate (AgNO3) to synthesize AgNPs, indicated by a color change from light to dark brown. Ultraviolet-visible (UV-vis) spectroscopy and transmission electron microscopy (TEM) were performed. Antibiofilm activity was measured by optical density (OD) against Enterococcus faecalis and Pseudomonas spp., while protein and cytoplasmic leakage assays assessed membrane damage. Antimicrobial activity and time-kill kinetics were tested against E. faecalis, Escherichia coli, Pseudomonas spp., and Staphylococcus aureus at 25– 100 μg/mL.
Results: UV-vis spectra showed a peak at 250–270 nm, confirming nanoparticle formation. TEM images revealed spherical particles. TC-AgNCs reduced biofilm OD from 0.525±0.015 to 0.455±0.005 (E. faecalis) and 0.54±0.015 to 0.46±0.005 (Pseudomonas spp.) at 25–100 μg/mL. Protein leakage increased from 0.34±0.006 to 0.38±0.006, and cytoplasmic leakage from 0.28±0.01 to 0.38±0.01. Antimicrobial activity was observed at 100 μg/mL: Pseudomonas spp. (22.5±1.0), E. faecalis (18.5±0.9), E. coli (16.0±0.8), and S. aureus (18.0±0.9). Time-kill analysis showed a reduction in bacterial counts with increasing concentration and time.
Conclusion: TC-AgNCs synthesized using G. superba extract are stable, predominantly spherical, and exhibit strong dose- and time-dependent antibacterial and antibiofilm activity, with membrane-disruptive effects against both Gram-positive and Gram-negative bacteria.
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References
1. Khan MA, Celik I, Khan HM, Shahid M, Shahzad A, Kumar S. Antibiofilm and anti-quorum sensing activity of Psidium guajava L. Leaf extract: In vitro and in silico approach. PLOS One. 2023;18(12):e0295524. doi: 10.1371/journal.pone.0295524, PMID 38113217
2. Paganelli FL, Van De Kamer T, Brouwer EC, Leavis HL, Woodford N, Bonten MJ. Lipoteichoic acid synthesis inhibition in combination with antibiotics abrogates growth of multidrug-resistant Enterococcus faecium. Int J Antimicrob Agents. 2017;49(3):355-63. doi: 10.1016/j. ijantimicag.2016.12.002, PMID 28188831
3. Chatterjee AK, Sarkar RK, Chattopadhyay AP, Aich P, Chakraborty R, Basu T. A simple robust method for synthesis of metallic copper nanoparticles of high antibacterial potency against E. coli. Nanotechnology. 2012;23(8):085103. doi: 10.1088/0957- 4484/23/8/085103, PMID 22293320
4. Ruparelia JP, Chatterjee AK, Duttagupta SP, Mukherji S. Strain specificity in antimicrobial activity of silver and copper nanoparticles. Acta Biomater. 2008;4(3):707-16. doi: 10.1016/j.actbio.2007.11.006, PMID 18248860
5. Gunalan S, Sivaraj R, Venckatesh R. Aloe barbadensis Miller mediated green synthesis of mono-disperse copper oxide nanoparticles: Optical properties. Spectrochim Acta A Mol Biomol Spectrosc. 2012;97:1140- 4. doi: 10.1016/j.saa.2012.07.096, PMID 22940049
6. Martinez-Gutierrez F, Olive PL, Banuelos A, Orrantia E, Nino N, Sanchez EM. Synthesis, characterization, and evaluation of antimicrobial and cytotoxic effect of silver and titanium nanoparticles. Nanomedicine. 2010;6(5):681-8. doi: 10.1016/j.nano.2010.02.001, PMID 20215045
7. Durán N, Durán M, De Jesus MB, Seabra AB, Fávaro WJ, Nakazato G. Silver nanoparticles: A new view on mechanistic aspects on antimicrobial activity. Nanomedicine. 2016;12(3):789-99. doi: 10.1016/j.nano.2015.11.016, PMID 26724539
8. Sasarom M, Wanachantararak P, Chaijareenont P, Okonogi S. Biosynthesis of copper oxide nanoparticles using Caesalpinia sappan extract: In vitro evaluation of antifungal and antibiofilm activities against Candida albicans. Drug Discov Ther. 2023;17(4):238-47. doi: 10.5582/ddt.2023.01032, PMID 37612046
9. Pollini M, Paladini F. The emerging role of silk fibroin for the development of novel drug delivery systems. Biomimetics (Basel). 2024;9(5):295. doi: 10.3390/biomimetics9050295, PMID 38786505
10. Gunasekaran T, Haile T, Nigusse T, Dhanaraju MD. Nanotechnology: An effective tool for enhancing bioavailability and bioactivity of phytomedicine. Asian Pac J Trop Biomed. 2014;4(Suppl 1):S1-7. doi: 10.12980/apjtb.4.2014c980, PMID 25183064
11. Rai M, Ingle AP, Birla S, Yadav A, Santos CA. Strategic role of selected noble metal nanoparticles in medicine. Crit Rev Microbiol. 2016;42(5):696-719. doi: 10.3109/1040841X.2015.1018131, PMID 26089024
12. Shanmugam, R., Munusamy, T., Jayakodi, S., Al-Ghanim, K. A., Nicoletti, M., Sachivkina, N., & Govindarajan, M. (2023). Probiotic-bacteria (Lactobacillus fermentum)-wrapped zinc oxide nanoparticles: biosynthesis, characterization, and antibacterial activity. Fermentation, 9(5), 413.
13. Yang J, Zhang L, He X, Gou X, Zong Z, Luo Y. In vitro and in vivo enhancement effect of glabridin on the antibacterial activity of colistin, against multidrug resistant Escherichia coli strains. Phytomedicine. 2024;130:155732. doi: 10.1016/j.phymed.2024.155732, PMID 38776738
14. Arasu MV, Al-Dhabi NA. Antibacterial activity of peptides and bio-safety evaluation: In vitro and in vivo studies against bacterial and fungal pathogens. J Infect Public Health. 2023;16(12):2031-7. doi: 10.1016/j.jiph.2023.09.006, PMID 37890227
15. Pelgrift RY, Friedman AJ. Nanotechnology as a therapeutic tool to combat microbial resistance. Adv Drug Deliv Rev. 2013;65(13- 14):1803-15. doi: 10.1016/j.addr.2013.07.011, PMID 23892192
16. Tan X, Huang Y, Rana A, Singh N, Abbey TC, Chen H. Optimization of an in vitro Pseudomonas aeruginosa biofilm model to examine antibiotic pharmacodynamics at the air-liquid interface. NPJ Biofilms Microbiomes. 2024;10(1):16. doi: 10.1038/s41522-024-00483-y, PMID 38429317
17. Jayakumar J, Vinod V, Arumugam T, Sathy BN, Biswas L, Kumar VA. Efficacy of lysostaphin functionalized silicon catheter for the prevention of Staphylococcus aureus biofilm. Int J Biol Macromol. 2024;256(2):128547. doi: 10.1016/j.ijbiomac.2023.128547, PMID 38048926
18. Lin H, Zhou R, Zhang M, Huang R, Fan C, Zhou S. In vitro antibacterial activity of a novel acid-activated antimicrobial peptide against Streptococcus mutans. Curr Protein Pept Sci. 2024;25(1):83-93. doi: 10 .2174/1389203724666230818111515, PMID 37594108
19. Wen Z, Chen C, Shang Y, Fan K, Li P, Li C. Baohuoside I inhibits virulence of multidrug-resistant Staphylococcus aureus by targeting the transcription Staphylococcus accessory regulator factor SarZ. Phytomedicine. 2024;130:155590. doi: 10.1016/j. phymed.2024.155590, PMID 38810547
20. Anishya D, Jain RK. Vanillin-mediated green-synthesised silver nanoparticles’ characterisation and antimicrobial activity: An in-vitro study. Cureus. 2024;16(1):e51659. doi: 10.7759/cureus.51659, PMID 38318582
21. Sahoo, B., Panigrahi, L. L., Jena, S., Jha, S., & Arakha, M. (2023). Oxidative stress generated due to photocatalytic activity of biosynthesized selenium nanoparticles triggers cytoplasmic leakage leading to bacterial cell death. RSC advances, 13(17), 11406-11414.
22. Azeem M, Siddique MH, Imran M, Zubair M, Mumtaz R, Younas M, Abdel-Maksoud MA. Assessing anticancer, antidiabetic, and antioxidant capacities in green-synthesized zinc oxide nanoparticles and solvent-based plant extracts. Heliyon. 2024;10(14):e34073.
23. Ahmajärvi, K. M., Isoherranen, K. M., Pessi, T. J., & Venermo, M. A. (2025). The Impact of Diagnostic Delay on Wound Healing—A Cohort Study in a Primary Care Setting. International wound journal, 22(5), e70141.
24. Pauline CR, Akshita TP, Kannan K, Sivaperumal P. Characterization and biological activity of silver nanoparticles from (Rhizophora mucronata) Mangrove extract. Nano Life. 2025;15(3):245001825. Jabeen N, Prabhalakshmi K, Dhanraj G, Ramasubburayan R. Biosynthesis of titanium dioxide nanoparticles using Sargassum tenerrimum as reductant and deciphering its antibiofilm role against cariogenic Candida albicans. Microb Pathog. 2025;202:107452. doi: 10.1016/j.micpath.2025.107452, PMID 40057005
26. Le NT, Thi TT, Ching YC, Nguyen NH, Nguyen DY, Truong QM. Garcinia mangostana shell and Tradescantia spathacea leaf extract-mediated one-pot synthesis of silver nanoparticles with effective antifungal properties. Curr Nanosci. 2021;17(5):762-71. doi: 10.2174/1 573413716666201222111244
27. Asha S, Asha A, Rajeshkumar S. Evaluation of phytochemical constituents and antimicrobial activity of silver nanoparticle synthesized Ipomoea nil against selected pathogens. Asian J Pharm Clin Res. 2017:183-7.
28. Nadeesh T, Dhara S, Manna P, Kayal P. Advanced nanotherapeutics in multiple sclerosis treatment: From blood-brain barrier crossing to remyelination enhancement. Int J Appl Pharm. 2026;18(1):71-83.
29. Manibhushanam S, Srinivasan UM, Narayanasamy D. Nanoparticle-embedded microneedles for enhanced transdermal delivery: Advances and applications. Int J Appl Pharm. 2026;18(1):20-32. doi: 10.22159/ ijap.2026v18i1.55869
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Copyright (c) 2026 Parivraj priyam, Renu Vajjiravelu, Rohit Kumar Singh, Dr Deepak Nallaswamy, Dr Santhoshkumar Jayakodi, Dr Rajeshkumar Shanmugam
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