Gene expression with Silver and Gold nanoparticles coated with amino acid on macrophages (RT-PCR)
RT-PCR Analysis on effect of Nanoparticles with free amino acids (l and d- tyrosine and tryptophan) on expression of inflammatory genes (TNF, il1b, and il10)
a) Tumor Necrosis Factor (TNF)
The treatment with L-TyrAgNPs resulted in the highest number of expression of TNF at a 4-hour interval, but that expression had a sharp decline in the second experiment which was left for 24 hours. A combination of nanoparticles together with free amino acids also shows a higher yield of TNF expression at a four hour compared to a 24-hour duration experiment
b) Interleukin 1 beta gene (IL1B) expression
In this experimentation, the best gene expression resulted in treatment with L- tryptophan combined with Silver (L- TrpAgNPs) followed by a combination of L-tyrosine with silver nanoparticles and D-Tyrosine recorded the lowest gene expression of IL1B at 4 hours. In the experiment left for 24 hours, the highest gene expression was at the control experiment followed by L-TrpAgNPs. This indicates a general decline of gene expression after 24-hour treatment with either free amino acids or a combination of silver nanoparticles.
c) Interleukin 10 gene (IL10)
A Combination of L- tryptophan and silver nanoparticles yielded the best gene expression results followed by a combination of L-tyrosine and silver nanoparticles at four hours. At 24 hour period the expression was highest at the control followed by the L-TrAgNPs. Slight expression was observed in treating the IL10 gene with free L and D- amino acids at during 4-hour experimentation. Peculiar results emerged after 24 hours indicating no expression with the treatment with free amino acids.
Inflammatory mediators such as TNF, IL10, and IL1B play a significant role in escalating inflammation (Yen, Hsu, and Tsai, 2009 and Münch, 2016). Nanoparticles such gold and silver nanoparticles have an increased expressed of these inflammatory mediators (Rosa, and Prockop, & White Healthcare and The Texas A&M University System, 2014). They stimulate the expression of such genes hence aggravating the problem of diseases of inflammation such as arthritis. Gold nanoparticles have greater gene expressed compared to silver nanoparticles, but in both cases, both have higher expression of genes of inflammatory mediators than the use of free amino acids alone (Watkins, Vasserot, Greene, Adams, Chiang, Zhang, Piehl, Hong, and He, Atyr Pharma, Inc., 2016). These particles get adsorbed into the cell membrane leading to their expression within the cell (Suresh, A.K., Pelletier, D.A., Wang, W., Morrell-Falvey, J.L., Gu, B. and Doktycz, M.J., 2012).
The use of nanoparticles indicates increased expression of genes of inflammatory mediators such as TNF, IL10, and IL1B hence should not be used on patients with inflammatory mediators.
dissertation, Technische Universität München).
Esophagus through IL-8 (KC) mediated inflammatory niche formation (Doctoral
Hong, F. and He, A., Atyr Pharma, Inc., 2016. Amino acyl tRNA synthetases for modulating inflammation. U.S. Patent 9,328,340.
Münch, N.S., 2016. High-fat diet accelerates esophageal dysplasia in a mouse model of Barrett
Rosa, R., Roddy, G.W. and Prockop, D.J., Scott & White Healthcare and The Texas A&M
Suresh, A.K., Pelletier, D.A., Wang, W., Morrell-Falvey, J.L., Gu, B. and Doktycz, M.J., 2012. Cytotoxicity induced by engineered silver nanocrystallites is dependent on surface coatings and cell types. Langmuir, 28(5), pp.2727-2735.
University System, 2014. Protein therapy for corneal inflammation, epithelial wound healing, and photoreceptor degeneration. U.S. Patent 8,759,298.
Watkins, J.D., Vasserot, A.P., Greene, L.A., Adams, R.A., Chiang, K.P., Zhang, W., Piehl, K.H.,
Yen, H.J., Hsu, S.H., and Tsai, C.L., 2009. Cytotoxicity and immunological response of gold and silver nanoparticles of different sizes. Small, 5(13), pp.1553-1561.