• info@icbcongress.com
• English
• Persian

• Login

Create Free Account

INTERNATIONAL CONGRESS OF

CancerGenetics

• Investigation on the effects of silver nanoparticles-attached to intracellular penetrating peptides on gene expression of NLRP3 in human monocyte-like macrophages

• Bita fazel,^1,* Romina Zamanikia,² Fatemeh ayazi,³ Sepide Moradkhani,⁴ Saman Hosseinkhani,⁵ Jalil Mehrzad,⁶
  1. Department of Immunology, Faculty of Veterinary Medicine, Tehran University, Tehran, Iran
  2. Department of Theriogenology , Faculty of Veterinary Medicine, Tehran University, Tehran, Iran
  3. Department of Immunology, Faculty of Veterinary Medicine, Tehran University, Tehran, Iran
  4. Department of Immunology, Faculty of Veterinary Medicine, Tehran University, Tehran, Iran
  5. Department of Biochemistry, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran
  6. saman_h@modares.ac.ir
  
  
  
• Introduction: Silver nanoparticles (AgNPs) are among the most widespread materials used in various fields, including therapeutic applications and everyday products such as those in the food industry. Considering their inevitable exposure it is then essential to conduct a comprehensive evaluation of the potential effects of AgNps on immune cells. Inflammation is initiated by the activation of different cells like monocytes and macrophages, which stimulate the inflammatory cytokines. Regulating the overexpression of inflammatory genes, such as NLRP3, could mitigate the potential negative impacts of the innate immune response. Studies show that chemically synthesized AgNPs are highly cytotoxic with low specificity. However, biosynthesis, particularly using plants like barberry, can significantly reduce its toxicity and the presence of unwanted compounds. Since the cell membrane serves as a barrier, nanocarriers should be designed to enhance the stability and permeability of drugs. In this study, conjugation of AgNPs with a cell-penetrating peptide (MiRGD) was performed to facilitate transportation and stimulate apoptosis in monocyte-like macrophages. Due to the positive charge of the peptide, the nanocarrier can specifically permeate into the target cell. Moreover, AgNPs have the potential to revolutionize cancer treatment by inducing apoptosis in cancer cells. Their functionalization with cell penetrates peptides (MiRGD) could increase the specificity towards cancer cells, with limited damage to healthy non-target tissues. With the development of drug delivery and inducing selective destruction of tumor cells, AgNPs might have significant potential in cancer therapies in the near future, especially in enhancing chemotherapy efficiency and reducing side effects.
• Methods: Blood samples were collected from healthy young individuals using tubes containing EDTA as anticoagulant. Blood mononuclear cells (PBMCs) were isolated using Ficoll and concentration gradient using a centrifuge at 2500 rpm for 20-25 minutes. The isolated mononuclear cells were suspended in an enriched RPMI medium. After harvesting monocytes from PBMCs, the monocytes attached to the bottom of the flask were subsequently separated. The monocyte-like macrophages were treated with different doses of green and green-synthesized AgNp conjugated with peptide and peptide alone. Cell viability was then evaluated by flow cytometry, staining with propidium iodide (PI), and annexin V labeled with fluorescein isothiocyanate (FITC). The purification of the Nano peptide and functionalization of the Nano-Ag coating with the Nano peptide were done based on the protocol provided faculty of Biological Science University of Tarbiat Modares . Finally, gene expression analysis was performed using real-time PCR.
• Results: Viability of monocyte-like macrophage cells treated with green AgNP conjugated with peptide tested with the minimum concentration (0.2μg/mL) for 24h illustrates a decline in apoptosis rate, suggesting the specificity of penetrating of nanocarrier into the cells. Lastly, Relative expression of the NLRP3 gene compared to control on human monocyte-macrophage was calculated based on the fold change method and results suggest that the samples treated with Ag peptide show a significant downregulation in NLRP3 gene, in comparison to AgNP treated cells.
• Conclusion: This study aims to provide insights into the significant down regulatory effects of green AgNP on anti-inflammatory and anti-apoptotic pathways. Moreover, the specificity of the nanocarrier's cellular penetration can be enhanced through the use of cell-penetrating peptides, which facilitate targeted delivery and contribute to a reduction in cytotoxicity in non-target cells. In summary, the findings of this study can provide a promising approach for the downregulation in inflammatory diseases.
• Keywords: green synthesis, peptide, inflammatory response, silver nanoparticle