After synthesizing silver nanoparticles (AgNPs) with sodium citrate as a reducing agent, an extensive examination of their biological activity is conducted. The antibacterial ability of AgNPs' was evaluated using microbial cultures, and the outcomes demonstrated that they significantly suppressed the growth of bacteria. The interactions between AgNPs and biological molecules are then clarified by biochemical experiments, which emphasize the modification of enzyme function and affinity for proteins and nucleic acids.

Additionally, alamarBlue dye is used in cell viability tests to assess the cytotoxic effects of AgNPs, revealing dose-dependent effects on mammalian cell lines. A reactive oxygen species (ROS) assay provided more about AgNPs' capacity to cause oxidative stress reduction in treated cells, offering understanding into the mechanisms behind their cytotoxicity as an anti-inflammatory agent. By using DCFDA dye, we can assay the relative level of ROS. The reduction of DCFDA dye by ROS indicate the level of ROS in a particular cell line.

The biological consequences of AgNPs, including their antibacterial activity, biochemical interactions, and cellular responses, are thoroughly understood thanks to this work. These results highlight the significance of evaluating AgNPs' safety and effectiveness in biological systems while also advancing the investigation of AgNPs for a range of biomedical and environmental application. Conducting studies on LPS induced inflammation in the K562 cell line and AgNps inhibitory activity on ROS may generate profound insights into therapy for inflammatory diseases such as rheumatoid Arthritis.

Keywords: Nanoparticle,Biochemical,Anti-Inflammatory,ROS,Mammalian cell

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1.  Cassano, D., Mapanao, A. K., Summa, M., Vlamidis, Y., Giannone, G., Santi, M. (2019). Biosafety and biokinetics of noble metals: The impact of their chemical nature.
2. ACS Applied Bio Materials, 2(10), 4464–4470. doi:10.1021/acsabm.9b00630. PMID 35021406. S2CID 204266885.
3. Singh, M., Singh, S., Prasad, S., & Gambhir, I. (2008). Nanotechnology in medicine and antibacterial effect of silver nanoparticles. Digest Journal of Nanomaterials and Biostructures, 3(3), 115-122.
4. Kalishwaralal, K., Deepak, V., Ramkumarpandian, S., Nellaiah, H., &Sangiliyandi, G. (2008). Extracellular biosynthesis of silver nanoparticles by the culture supernatant of Bacillus licheniformis. Materials Letters, 62(28), 4411-4413.
5. Chen, X., &Schluesener, H. J. (2008). Nanosilver: A nanoproduct in medical application. Toxicology Letters, 176(1), 1–12.
6. Morones, J. R., Elechiguerra, J. L., Camacho, A., Holt, K., Kouri, J. B., Ramírez, J. T., & Yacaman, M. J. (2005). The bactericidal effect of silver nanoparticles. Nanotechnology, 16(10), 2346–2353.
7. Rai, M., Yadav, A., & Gade, A. (2009). Silver nanoparticles as a new generation of antimicrobials. Biotechnology Advances, 27(1), 76–83.
8. Skrabalak, S. E., Chen, J., Sun, Y., Lu, X., Au, L., Cobley, C. M., & Xia, Y. (2008). Gold nanocages: Synthesis, properties, and applications. Accounts of Chemical Research, 41(12), 1587–1595.
9. Galatage, S. T., Hebalkar, A. S., Dhobale, S. V., Mali, O. R., Kumbhar, P. S., Nikade, S. V., &Killedar, S. G. (2021). Green strategy–based synthesis of silver nanoparticles for antibacterial applications Frontiers in Nanotechnology, 3(2021). https://doi.org/10.3389/fnano.2021.697303
10. Gamboa, S. M., Rojas, E. R., Martínez, V. V., et al. (2019). Synthesis and characterization of silver nanoparticles and their application as an antibacterial agent. International Journal of Biosensors & Bioelectronics, 5(5), 166-173. doi: 10.15406/ijbsbe.2019.05.00172
11. Kim, J. H., Kuk, E., Yu, K. N., Kim, J. H., Park, S. J., Lee, H. J., ... & Cho, M. H. (2007). Antimicrobial effects of silver nanoparticles. Nanomedicine: Nanotechnology, Biology and Medicine, 3 (1), 95-101.
12. Jung, M. R., Lee, J. Y., Park, C. B., & Kim, B. C. (2015). Efficient synthesis of silver nanoparticles using a ceramic heater with a porous membrane. Journal of Nanoparticle Research, 17(8), 324.
13. Amendola, V., & Meneghetti, M. (2009). Laser ablation synthesis in solution and size manipulation of noble metal nanoparticles. Physical Chemistry Chemical Physics, 11(18), 3805-3821.
14. Amendola, V., Polizzi, S., & Meneghetti, M. (2015). Free silver nanoparticles synthesized by laser ablation in organic solvents and their easy functionalization. Langmuir, 31(17), 4804-4811.
15. Li, X., Zhang, S., & Kulinich, S. A. (2015). Formation of colloidal silver nanoparticles stabilized by Na+-poly (γ-glutamic acid)-silver nitrate complex via chemical reduction process. Colloids and Surfaces A: Physicochemical and Engineering Aspects, 468, 88-94.
16. Durán, N., Durán, M., de Jesus, M. B., Seabra, A. B., Fávaro, W. J., & Nakazato, G. (2016). Silver nanoparticles: A new view on mechanistic aspects on antimicrobial activity. Nanomedicine: Nanotechnology, Biology, and Medicine, 12(3), 789-799.
17. Pal, S., Tak, Y. K., & Song, J. M. (2007). Does the antibacterial activity of silver nanoparticles depend on the shape of the nanoparticle? A study of the Gram-negative bacterium Escherichia coli. Applied and Environmental Microbiology, 73(6), 1712-1720.
18. Sondi, I., & Salopek-Sondi, B. (2004). Silver nanoparticles as antimicrobial agent: A case study on E. coli as a model for Gram-negative bacteria. Journal of Colloid and Interface Science, 275(1), 177-182.
19. Kim, J. S., Kuk, E., Yu, K. N., et al. (2007). Antimicrobial effects of silver nanoparticles. Nanomedicine: Nanotechnology, Biology, and Medicine, 3(1), 95-101.
20. Zhao, G., & Stevens, S. E. (1998). Multiple parameters for thecomprehensive evaluation of the susceptibility of Escherichia coli to the silver ion. Bio Metals, 11,27-32
21. Hulkoti, N. I., & Taranath, T. C. (2017). Influence of physico- chemical parameters on the fabrication of silver nanoparticles using Petrea volubilis L. stem broth and its antimicrobial efficacy. International Journal of Pharmaceutical Sciences and DrugResearch, 9, 72–78
22. Sohn, E. K., Johari, S. A., Kim, T. G., Kim, J. K., Kim, E., & Lee, J. H. (2015). Aquatic toxicity comparison of silver nanoparticles and silver nanowires. Bio Med Research International, 2015,893049
23. Capek, I. (2004). Preparation of metal nanoparticles in water-in-oil (w/o) microemulsions. Advances in colloid and interface science, 110,49-74
24. Hutchison, J. E., & Greener. (2008). Nanoscience: A proactive approach to advancing applications and reducing implications of nanotechnology. ACS Nano,2 395-402
25. Zewde, B., Ambaye, A., Stubbs III, J., & Raghavan, D. (2016). A review of stabilized silver nanoparticles–synthesis, biological properties, characterization, and potential areas of applications.Nanomedicine,4(1043),1-4
26. Berry CC, Curtis ASG. Functionalization of magnetic nanoparticles for applications in biomedicine. J Phys D: Appl Phys. 2003;36:R198. https://doi.org/10.1088/0022-3727/36/13/203 2.
27. Galatage ST, Hebalkar AS, Dhobale SV, Mali OR, Kumbhar PS, Nikade SV, Killedar SG. Silver Nanoparticles: Properties. Synthesis, Characterization, Applications and Future Trends, Silver Micro-Nanoparticles–Properties, Synthesis, Characterization, and Applications, IntechOpen. 2021.
28. Skrabalak SE, Wiley BJ, Kim M, Formo EV, Xia Y. On the polyol synthesis of silver nanostructures: glycolaldehyde as a reducing agent. Nano letters. 2008 Jul 9;8(7):2077-81.
29. Rai M, Gade A, Yadav A. Biogenic nanoparticles: an introduction to what they are, how they are synthesized and their applications. InMetal nanoparticles in microbiology 2011 Mar 4 (pp. 1-14). Berlin, Heidelberg: Springer Berlin Heidelberg.
30. Yokoyama K, Welchons DR. The conjugation of amyloid beta protein on the gold colloidal nanoparticles’ surfaces. Nanotechnology. 2007 Jan 31;18(10):105101.
31. Saxena A, Tripathi RM, Zafar F, Singh P. Green synthesis of silver nanoparticles using aqueous solution of Ficus benghalensis leaf extract and characterization of their antibacterial activity. Materials letters. 2012 Jan 15;67(1):91-4.
32. Khalil MM, Ismail EH, El-Baghdady KZ, Mohamed D. Green synthesis of silver nanoparticles using olive leaf extract and its antibacterial activity. Arabian Journal of chemistry. 2014 Dec 1;7(6):1131-9.
33. Rai M, Yadav A, Gade A. Silver nanoparticles as a new generation of antimicrobials. Biotechnology advances. 2009 Jan 1;27(1):76-83.

After synthesizing silver nanoparticles (AgNPs) with sodium citrate as a reducing agent, an extensive examination of their biological activity is conducted. The antibacterial ability of AgNPs' was evaluated using microbial cultures, and the outcomes demonstrated that they significantly suppressed the growth of bacteria. The interactions between AgNPs and biological molecules are then clarified by biochemical experiments, which emphasize the modification of enzyme function and affinity for proteins and nucleic acids.

Additionally, alamarBlue dye is used in cell viability tests to assess the cytotoxic effects of AgNPs, revealing dose-dependent effects on mammalian cell lines. A reactive oxygen species (ROS) assay provided more about AgNPs' capacity to cause oxidative stress reduction in treated cells, offering understanding into the mechanisms behind their cytotoxicity as an anti-inflammatory agent. By using DCFDA dye, we can assay the relative level of ROS. The reduction of DCFDA dye by ROS indicate the level of ROS in a particular cell line.

The biological consequences of AgNPs, including their antibacterial activity, biochemical interactions, and cellular responses, are thoroughly understood thanks to this work. These results highlight the significance of evaluating AgNPs' safety and effectiveness in biological systems while also advancing the investigation of AgNPs for a range of biomedical and environmental application. Conducting studies on LPS induced inflammation in the K562 cell line and AgNps inhibitory activity on ROS may generate profound insights into therapy for inflammatory diseases such as rheumatoid Arthritis.

Keywords: Nanoparticle,Biochemical,Anti-Inflammatory,ROS,Mammalian cell

🔒 This is a subscription article

Full text is available to subscribers and institutional members. Please choose an option below to access it.

1.  Cassano, D., Mapanao, A. K., Summa, M., Vlamidis, Y., Giannone, G., Santi, M. (2019). Biosafety and biokinetics of noble metals: The impact of their chemical nature.
2. ACS Applied Bio Materials, 2(10), 4464–4470. doi:10.1021/acsabm.9b00630. PMID 35021406. S2CID 204266885.
3. Singh, M., Singh, S., Prasad, S., & Gambhir, I. (2008). Nanotechnology in medicine and antibacterial effect of silver nanoparticles. Digest Journal of Nanomaterials and Biostructures, 3(3), 115-122.
4. Kalishwaralal, K., Deepak, V., Ramkumarpandian, S., Nellaiah, H., &Sangiliyandi, G. (2008). Extracellular biosynthesis of silver nanoparticles by the culture supernatant of Bacillus licheniformis. Materials Letters, 62(28), 4411-4413.
5. Chen, X., &Schluesener, H. J. (2008). Nanosilver: A nanoproduct in medical application. Toxicology Letters, 176(1), 1–12.
6. Morones, J. R., Elechiguerra, J. L., Camacho, A., Holt, K., Kouri, J. B., Ramírez, J. T., & Yacaman, M. J. (2005). The bactericidal effect of silver nanoparticles. Nanotechnology, 16(10), 2346–2353.
7. Rai, M., Yadav, A., & Gade, A. (2009). Silver nanoparticles as a new generation of antimicrobials. Biotechnology Advances, 27(1), 76–83.
8. Skrabalak, S. E., Chen, J., Sun, Y., Lu, X., Au, L., Cobley, C. M., & Xia, Y. (2008). Gold nanocages: Synthesis, properties, and applications. Accounts of Chemical Research, 41(12), 1587–1595.
9. Galatage, S. T., Hebalkar, A. S., Dhobale, S. V., Mali, O. R., Kumbhar, P. S., Nikade, S. V., &Killedar, S. G. (2021). Green strategy–based synthesis of silver nanoparticles for antibacterial applications Frontiers in Nanotechnology, 3(2021). https://doi.org/10.3389/fnano.2021.697303
10. Gamboa, S. M., Rojas, E. R., Martínez, V. V., et al. (2019). Synthesis and characterization of silver nanoparticles and their application as an antibacterial agent. International Journal of Biosensors & Bioelectronics, 5(5), 166-173. doi: 10.15406/ijbsbe.2019.05.00172
11. Kim, J. H., Kuk, E., Yu, K. N., Kim, J. H., Park, S. J., Lee, H. J., ... & Cho, M. H. (2007). Antimicrobial effects of silver nanoparticles. Nanomedicine: Nanotechnology, Biology and Medicine, 3 (1), 95-101.
12. Jung, M. R., Lee, J. Y., Park, C. B., & Kim, B. C. (2015). Efficient synthesis of silver nanoparticles using a ceramic heater with a porous membrane. Journal of Nanoparticle Research, 17(8), 324.
13. Amendola, V., & Meneghetti, M. (2009). Laser ablation synthesis in solution and size manipulation of noble metal nanoparticles. Physical Chemistry Chemical Physics, 11(18), 3805-3821.
14. Amendola, V., Polizzi, S., & Meneghetti, M. (2015). Free silver nanoparticles synthesized by laser ablation in organic solvents and their easy functionalization. Langmuir, 31(17), 4804-4811.
15. Li, X., Zhang, S., & Kulinich, S. A. (2015). Formation of colloidal silver nanoparticles stabilized by Na+-poly (γ-glutamic acid)-silver nitrate complex via chemical reduction process. Colloids and Surfaces A: Physicochemical and Engineering Aspects, 468, 88-94.
16. Durán, N., Durán, M., de Jesus, M. B., Seabra, A. B., Fávaro, W. J., & Nakazato, G. (2016). Silver nanoparticles: A new view on mechanistic aspects on antimicrobial activity. Nanomedicine: Nanotechnology, Biology, and Medicine, 12(3), 789-799.
17. Pal, S., Tak, Y. K., & Song, J. M. (2007). Does the antibacterial activity of silver nanoparticles depend on the shape of the nanoparticle? A study of the Gram-negative bacterium Escherichia coli. Applied and Environmental Microbiology, 73(6), 1712-1720.
18. Sondi, I., & Salopek-Sondi, B. (2004). Silver nanoparticles as antimicrobial agent: A case study on E. coli as a model for Gram-negative bacteria. Journal of Colloid and Interface Science, 275(1), 177-182.
19. Kim, J. S., Kuk, E., Yu, K. N., et al. (2007). Antimicrobial effects of silver nanoparticles. Nanomedicine: Nanotechnology, Biology, and Medicine, 3(1), 95-101.
20. Zhao, G., & Stevens, S. E. (1998). Multiple parameters for thecomprehensive evaluation of the susceptibility of Escherichia coli to the silver ion. Bio Metals, 11,27-32
21. Hulkoti, N. I., & Taranath, T. C. (2017). Influence of physico- chemical parameters on the fabrication of silver nanoparticles using Petrea volubilis L. stem broth and its antimicrobial efficacy. International Journal of Pharmaceutical Sciences and DrugResearch, 9, 72–78
22. Sohn, E. K., Johari, S. A., Kim, T. G., Kim, J. K., Kim, E., & Lee, J. H. (2015). Aquatic toxicity comparison of silver nanoparticles and silver nanowires. Bio Med Research International, 2015,893049
23. Capek, I. (2004). Preparation of metal nanoparticles in water-in-oil (w/o) microemulsions. Advances in colloid and interface science, 110,49-74
24. Hutchison, J. E., & Greener. (2008). Nanoscience: A proactive approach to advancing applications and reducing implications of nanotechnology. ACS Nano,2 395-402
25. Zewde, B., Ambaye, A., Stubbs III, J., & Raghavan, D. (2016). A review of stabilized silver nanoparticles–synthesis, biological properties, characterization, and potential areas of applications.Nanomedicine,4(1043),1-4
26. Berry CC, Curtis ASG. Functionalization of magnetic nanoparticles for applications in biomedicine. J Phys D: Appl Phys. 2003;36:R198. https://doi.org/10.1088/0022-3727/36/13/203 2.
27. Galatage ST, Hebalkar AS, Dhobale SV, Mali OR, Kumbhar PS, Nikade SV, Killedar SG. Silver Nanoparticles: Properties. Synthesis, Characterization, Applications and Future Trends, Silver Micro-Nanoparticles–Properties, Synthesis, Characterization, and Applications, IntechOpen. 2021.
28. Skrabalak SE, Wiley BJ, Kim M, Formo EV, Xia Y. On the polyol synthesis of silver nanostructures: glycolaldehyde as a reducing agent. Nano letters. 2008 Jul 9;8(7):2077-81.
29. Rai M, Gade A, Yadav A. Biogenic nanoparticles: an introduction to what they are, how they are synthesized and their applications. InMetal nanoparticles in microbiology 2011 Mar 4 (pp. 1-14). Berlin, Heidelberg: Springer Berlin Heidelberg.
30. Yokoyama K, Welchons DR. The conjugation of amyloid beta protein on the gold colloidal nanoparticles’ surfaces. Nanotechnology. 2007 Jan 31;18(10):105101.
31. Saxena A, Tripathi RM, Zafar F, Singh P. Green synthesis of silver nanoparticles using aqueous solution of Ficus benghalensis leaf extract and characterization of their antibacterial activity. Materials letters. 2012 Jan 15;67(1):91-4.
32. Khalil MM, Ismail EH, El-Baghdady KZ, Mohamed D. Green synthesis of silver nanoparticles using olive leaf extract and its antibacterial activity. Arabian Journal of chemistry. 2014 Dec 1;7(6):1131-9.
33. Rai M, Yadav A, Gade A. Silver nanoparticles as a new generation of antimicrobials. Biotechnology advances. 2009 Jan 1;27(1):76-83.