The effects of indirect exposure of nanosilver on caspase-8 and caspase-9 levels in liver and brain of suckling rats

Document Type: Research Paper


Department of Biology, Falavarjan Branch, Islamic Azad University, Isfahan, Iran


Objective(s): The adverse health effects of nanosilver (AgNp) on adult animal models have been well documented. However, data is scarce regarding the toxic effects of AgNp on sensitive developmental stages. The present study aimed to investigate the effects of maternal milk exposure to AgNp on apoptosis induction in the liver and brain of the offspring of rats.
Materials and Methods: Lactating Wistar rats were intragastrically exposed to the vehicle (deionized water) or two doses of AgNp (25 and 100 mg/kg) for 21 days. Liver and brain samples were collected from the male pups of the mothers on postnatal day 21. The silver content and levels of caspase-8 and caspase-9 in the tissues were measured using the ICP-MS analysis and ELISA assay, respectively. For histopathological examinations, the tissue sections were stained using the hematoxylin-eosin (H&E) stain and examined by light microscopy.
Results: A significant, dose-dependent increase was observed in the silver content of the liver and brain of the pups and maternal milk exposed to AgNp. In addition, the level of caspase-9 significantly increased in the liver and brain in the pups exposed to the high dose of AgNp (100 mg/kg-1), while no significant changes were observed in the level of caspase-8 in the experimental groups compared to the controls. Histopathological studies also demonstrated tissue damage in the liver and brain of the pups exposed to the high dose of AgNp.
Conclusion: According to the results, lactational exposure to AgNp may induce apoptosis via the intrinsic pathway in the offspring tissues of rats. However, further investigation is required in order to document these findings.


1.Ahamed M, Al Salhi MS, Siddiqui MKJ. Silver nanoparticle applications and human health. Clinica Chimica Acta. 2010; 411: 1841-1848.
2.Pulit J, Banach M, Kowalski Z. Nanosilver making difficult decisions. Ecol Chem Eng. 2011; 18(2): 185-196.
3.Ciftci H, Turk M, Tamer U, Karahan S. Silver nanoparticles: cytotoxic, apoptotic, and necrotic effects on MCF-7 cells. Turkish J Biol. 2013; 37: 573-581.
4.Gaillet S, Rouanet JM. Silver nanoparticles: their potential toxic effects after oral exposure and underlying mechanisms–a review. Food Chem Toxicol. 2015; 77: 58-63.
5.Ahlberg S, Antonopulos A, Diendorf J, Dringen R, Epple M, Flöck R, Goedecke W, Graf C, Haberl N, Helmlinger J, Herzog F, Heuer F, Hirn S, Johannes C, Kittler S, Köller M, Korn K, Kreyling WG, Krombach F, Lademann J, Loza K, Luther EM, Malissek M, Meinke MC, Nordmeyer D, Pailliart A, Raabe J, Rancan F, Rothen-Rutishauser B, Rühl E, Schleh C, Seibel A, Sengstock C, Treuel L, Vogt A, Weber K, Zellner R. PVP-coated, negatively charged silver nanoparticles: A multi-center study of their physicochemical characteristics, cell culture and in vivo experiments. Beilstein J Nanotechnol. 2014; 5: 1944-65.
6.Hadrup N, Lam HR. Oral toxicity of silver ions, silver nanoparticles and colloidal silver–a review. Regul Toxicol Pharmacol. 2014; 68: 1-7.
7.Johnston HJ, Hutchison G, Christensen FM, Peters S, Hankin S, Stone V. A review of the in vivo and in vitro toxicity of silver and gold particulates: particle attributes and biological mechanisms responsible for the observed toxicity. Crit Rev Toxicol. 2010; 40(4): 328-346.
8.Govender A, Phulukdaree A, Gengan RM, Anand K, Chuturgoon AA. Silver nanoparticles of Albizia adianthifolia: the induction of apoptosis in human lung carcinoma cell line. J Nanobiotech. 2013; 11(5): 1-9.
9.Zhang T, Wang L, Chen Q, Chen C. Cytotoxic potential of silver nanoparticles. Yonsei Med J. 2014; 55(2): 283-291.
10.Kim TH, Kim M, Park HS, Shin US, Gong MS, Kim HW. Size-dependent cellular toxicity of silver nanoparticles. J Biomed Mater Res A. 2012; 100 (4):1033-1043.
11.Kim SH, Ko JW, Koh SK, Lee IC, Jung-Mo Son JM, Moon C, Kim, Dong SH, Shin H, Kim JC. Silver nanoparticles induce apoptotic cell death in cultured cerebral cortical neurons. Mol Cell Toxicol. 2014; 10(2): 173-179.
12. Zhao X, Ren X, Zhu R, Luo Z, Ren B . Zinc oxide nanoparticles induce oxidative DNA damage and ROS-triggered mitochondria-mediated apoptosis in zebrafish embryos. Aquat Toxicol. 2016; 180: 56-70.
13.Andon FT, Fadeel B. Programmed cell death: molecular mechanisms and implications for safety assessment of nanomaterials. Acc Chem Res. 2013; 46(3):733–742.
14.Guicciardi ME, Gores GJ. Apoptosis: a mechanism of acute and chronic liver injury. Recent adv basic sci. 2005; 54: 1024-1033.
15.Rastogi RP, SinhaRP. Apoptosis: molecular mechanisms and pathogenicity. Excli J. 2009; 8: 155-181.
16.Oskarsson A, Hallén IP, Sundberg J, Grawé KP. Risk assessment in relation to neonatal metal exposure. Analyst. 1998; 123: 19-23.
17.Melnik EA, Buzulukov YP, Demin VF, Gmoshinski IV, Tyshko NV, Tutelyan VA. Transfer of silver nanoparticles through the placenta and breast milk during in vivo experiments on rats. Acta naturae. 2013; 5(3): 107-115.
18.Zhang C, Zhai S, Wu L, Bai Y, Jia J, Zhang Y, Zhang B, Yan B. Induction of size-dependent breakdown of blood-milk barrier in lactating mice by TiO2 nanoparticles. PLoS ONE. 2015; 10(4): 1-18.
19.Takeda K, Suzuki K, Ishihara A, Kubo-Irie M, Fujimoto R, Tabata M, Oshio S, Nihei Y, Ihara T, Sugamata M . Nanoparticles transferred from pregnant mice to their offspring can damage the genital and cranial nerve systems. J Health Sci. 2009; 55(1): 95-102.
20.Kulvietis V, Zalgeviciene V, Didziapetriene J, Rotomskis R. Transport of nanoparticles through the placental barrier. Tohoku J Exp Med. 2011; 225(4): 225-234.
21.Tiwari DK, Jin T, Behari J. Dose-dependent in-vivo toxicity assessment of silver nanoparticle in Wistar rats. Toxicol Mech Methods. 2011; 21(1): 13-24.
22.Lee TY, Liu MS, Huang LJ Lue SI, LinLC, Kwan AL, Yang RC. Bioenergetics’ failure correlates with autophagy and apoptosis in rat liver following silver nanoparticle intraperitoneal administration. Part Fibre Toxicol. 2013; 10(40):1-13.
23.MorishitaY, Yoshioka Y, Takimura Y, Shimizu Y, Namba Y, Nojiri N, Ishizaka T, Takao K, Yamashita F, Takuma K, Ago Y, Nagano K, Mukai Y, Kamada H, Tsunoda S, Saito S, Matsuda T, Hashida M, Miyakawa T, Higashisaka K, Tsutsumi Y. Distribution of Silver Nanoparticles to Breast Milk and Their Biological Effects on Breast-Fed Offspring Mice. ACS Nano. 2017; 10(9): 8181-8191.
24.Mc Manaman JL, Neville MC. Mammary physiology and milk secretion. Adv Drug Deliv Rev . 2013; 55(5): 629-641.
25.Sumner SCJ, Fennell TR, Snyder RW, Taylor GF, Lewin AH . Distribution of carbon-14 labeled C60 (C-14 C60) in the pregnant and in the lactating dam and the effect of C60 exposure on the biochemical profile of urine. J Appl Toxicol. 2010; 30(4): 354-360.
26.Borm PJ, Kreyling WJ. Toxicological hazards of inhaled nanoparticles potential implications for drug delivery. J Nanosci Nanotechnol. 2004; 4(5): 521-531.
27.Sharma HS, Ali SF, Hussain SM, Schlager JJ, Sharma A. Influence of engineered nanoparticles from metals on the blood-brain barrier permeability, cerebral blood flow, brain edema and neurotoxicity. An experimental study in the rat and mice using biochemical and morphological approaches. J Nanosci Nanotechnol. 2009; 9(8): 5055-5072.
28.Feng X, Chen A, Zhang Y, Wang J, Shao L, Wei L. Central nervous system toxicity of metallic nanoparticles. Int J Nanomedicine. 2015; 10: 4321-4340.
29.Kim YS, Kim JS, Cho HS, Rha DS, Kim JM, Park JD, Choi BS, Lim R, Chang HK, Chung YH, Kwon IH, Jeong J, Han BS, Yu IJ. Twenty-eight-day oral toxicity, genotoxicity, and gender-related tissue distribution of silver nanoparticles in Sprague dawley rats. Inhal Toxicol. 2008; 20(6): 575-583.
30.Cha K, Hong HW, Choi YG, Lee MJ, Park JH, Chae HK, Ryu G, Myung H. Comparison of acute responses of mice livers to short-term exposure to nano-sized or micro-sized silver particles. Biotechnol Lett. 2008; 30(11): 1893-1899.
31.Heydarnejad MS, Yarmohammadi-Samani P, Mobini Dehkordi M, Shadkhast M, Rahnama S. Histopathological effects of nanosilver (Ag-NPs) in liver after dermal exposure during wound healing. Nanomed J. 2014; 1(3): 191-197.
32.Xu F, Piett C, Farkas S, Qazzaz M, Syed NI. Silver nanoparticles (AgNPs) cause degeneration of cytoskeleton and disrupt synaptic machinery of cultured cortical neurons. Mol Brain. 2013; 6(29): 1-15.
33. Skalska J, Frontczak-Baniewicz M, Strużyńska L. Synaptic degeneration in rat brain after prolonged oral exposure to silver nanoparticles. Neurotoxicology. 2015; 46: 145–154.
34. Chen X, Schluesener HJ. Nanosilver: A nanoproduct in medical application. Toxicol Lett. 2008; 176(1): 1-12.
35.Fatemi M, Hayatiroodbari N, Ghaedi K, Naderi Gh. The effects of prenatal exposure to silver nanoparticles on the developing brain in neonatal rats. J Biol Res. 2013; 20: 233-242.
36.Hsin Y, Chen C, Huang S, Shih TS, Lai PS, Chueh PJ . The apoptotic effect of nanosilver is mediated by ROS and JNK-dependent mechanism involving the mitochondrial pathway in NIH3T3 cells. Toxicol Lett. 2008; 179(3): 130-139.
37.Zhu B, Li Y, Lin Z, Zhao M, Xu T, Wang C, Deng N. Silver Nanoparticles Induce HePG-2 Cells Apoptosis through ROS-Mediated Signaling Pathways. Nanoscale Res Lett. 2016; 11(198): 1-8.