A Low Dose of Nanoparticulate Silver Induces Mitochondrial Dysfunction and Autophagy in Adult Rat Brain.

Extensive incorporation of silver nanoparticles (AgNPs) into many medical and consumer products has raised concerns about biosafety. Since nanosilver accumulates persistently in the central nervous system, it is important to assess its neurotoxic impacts. We investigated a model of prolonged exposure of adult rats to a low environmentally relevant dose of AgNPs (0.2 mg/kg b.w.). Ultrastructural analysis revealed pathological alterations in mitochondria such as swelling and cristolysis. Besides, elongated forms of mitochondria were present. Level of adenosine triphosphate was not altered after exposure, although a partial drop of mitochondrial membrane potential was noted. Induction of autophagy with only early autophagic forms was observed in AgNP-exposed rat brains as evidenced by ultrastructural markers. Increased expression of two protein markers of autophagy, beclin 1 and microtubule-associated proteins 1A/1B light chain 3B (MAP LC3-II), was observed, indicating induction of autophagy. Expression of lysosome-related Rab 7 protein and cathepsin B did not change, suggesting inhibition of physiological flux of autophagy. Our results show that exposure to a low, environmentally relevant dose of AgNPs leads to induction of autophagy in adult rat brain in response to partial mitochondrial dysfunction and to simultaneous interfering with an autophagic pathway. The cell compensates for the defective autophagy mechanism via development of enhanced mitochondrial biodynamic.

Ultrastructural and biochemical features of cerebral microvessels of adult rat subjected to a low dose of silver nanoparticles.

The widespread use of silver nanoparticles (AgNPs) in medicine and in multiple commercial products has motivated researchers to investigate their potentially hazardous effects in organisms. Since AgNPs may easily enter the brain through the blood-brain barrier (BBB), characterization of their interactions with cellular components of the neurovascular unit (NVU) is of particular importance. Therefore, in an animal model of prolonged low-dose exposure, we investigate the extent and mechanisms of influence of AgNPs on cerebral microvessels. Adult rats were treated orally with small (10 nm) AgNPs in a dose of 0.2 mg/kg b.w. over a 2-week period. A silver citrate-exposed group was established as a positive control of ionic silver effects. Alterations in the expression of tight junction proteins claudin-5, ZO-1, and occludin, were observed. These effects are accompanied by ultrastructural features indicating enhanced permeability of microvessels such as focal edema of perivascular astrocytic processes and surrounding neuropil. We did not identify changes in the expression of PDGFßR which is a marker of pericytes. Ultrastructural alterations in these cells were not identified. The results show that altered integrity of cerebral vessels under a low-dose of AgNP-exposure may be the consequence of dysfunction of endothelial cells caused by disruption of tight junction proteins.

Mechanisms Underlying Neurotoxicity of Silver Nanoparticles.

The potent antimicrobial properties of nanoparticulate silver (AgNPs) have led to broad interest in using them in a wide range of commercial and medical applications. Although numerous in vivo and in vitro studies have provided evidence of toxic effects, rapid commercialization of AgNP-based nanomaterials has advanced without characterization of their potential environmental and health hazards. There is evidence that AgNPs can be translocated from the blood to the brain, regardless the route of exposure, and accumulate in the brain over time. As the brain is responsible for basic physiological functions and controls all human activities, it is important to assess the hazardous influence of AgNPs released from widely used nanoproducts and possible side effects of AgNP-based therapies. A number of studies have suggested that the size, shape and surface coating, as well as rates of silver ion release and interactions with proteins are the key factors determining the neurotoxicity of AgNPs. AgNPs target endothelial cells forming the blood-brain barrier, neurons and glial cells and leads finally to oxidative stress-related cell death. In this chapter, we review in detail current data on the impact of AgNPs on the central nervous system and discuss the possible mechanisms of their neurotoxic effects.

Oxidative stress in rat brain but not in liver following oral administration of a low dose of nanoparticulate silver.

While it is known that silver nanoparticles (AgNPs) can enter the brain, our knowledge of AgNP-induced neurotoxicity remains incomplete. We investigated the ability of 10 nm citrate-stabilized AgNPs to generate oxidative stress in brain and liver of adult male Wistar rats after repeated oral exposure for 14 days, using a low dose of 0.2 mg/kg b.w. as compared with the same dose of ionic silver (silver citrate). In AgNP-exposed animals, the level of reactive oxygen species (ROS), lipid peroxidation (MDA) and glutathione peroxidase (GPx) activity were found to be significantly higher in brain relative to the control group receiving saline. Administration of ionic silver (silver citrate) increased ROS and MDA levels in both tissues. Activities of GPx in brain so as superoxide dismutase (SOD) and catalase (CAT) in liver of exposed animals were also elevated. Besides, AgNPs and silver ions were both found to cause statistically significant decrease in the reduced-to-oxidized glutathione ratio (GSH/GSSG) in brain. The results show that exposure to a very low dose of particulate silver generates mild oxidative stress in the brain but not in the liver of rats, indicating a role of oxidative stress in AgNP-induced neurotoxicity.

Influence of a low dose of silver nanoparticles on cerebral myelin and behavior of adult rats.

Nanoscale particles have large surface to volume ratio that significantly enhances their chemical and biological reactivity. Although general toxicity of nano silver (nanoAg) has been intensively studied in both in vitro and in vivo models, its neurotoxic effects are poorly known, especially those of low-dose exposure. In the present study we assess whether oral administration of nanoAg influences behavior of exposed rats and induces changes in cerebral myelin. We examine the effect of prolonged exposure of adult rats to small (10nm) citrate-stabilized nanoAg particles at a low dose of 0.2mg/kg b.w. (as opposed to the ionic silver) in a comprehensive behavioral analysis. Myelin ultrastructure and the expression of myelin-specific proteins are also investigated. The present study reveals slight differences with respect to behavioral effects of Ag(+)- but not nanoAg-treated rats. A weak depressive effect and hyperalgesia were observed after Ag(+) exposure whereas administration of nanoAg was found to specifically increase body weight and body temperature of animals. Both nanoAg and Ag(+) induce morphological disturbances in myelin sheaths and alter the expression of myelin-specific proteins CNP, MAG and MOG. These results suggest that the CNS may be a target of low-level toxicity of nanoAg.

Synaptic degeneration in rat brain after prolonged oral exposure to silver nanoparticles.

Neurotoxicity of silver nanoparticles has been confirmed in both in vitro and in vivo studies. However, the mechanisms of the toxic action have not been fully clarified. Since nanoparticles are likely to have the ability to enter the brain and significantly accumulate in this organ, it is important to investigate their neurotoxic mechanisms. Here we examine the effect of prolonged exposure of rats to small (10nm) citrate-stabilized silver nanoparticles (as opposed to the ionic silver) on synapse ultrastructure and specific proteins. Administration of both nanosilver and ionic silver over a two-week period resulted in ultrastructural changes including blurred synapse structure and strongly enhanced density of synaptic vesicles clustering in the center of the presynaptic part. Disturbed synaptic membrane leading to liberation of synaptic vesicles into neuropil, which testifies for strong synaptic degeneration, was characteristic feature observed under AgNPs exposure. Also a noteworthy finding was the presence of myelin-like structures derived from fragmented membranes and organelles which are associated with neurodegenerative processes. Additionally, we observed significantly decreased levels of the presynaptic proteins synapsin I and synaptophysin, as well as PSD-95 protein which is an indicator of postsynaptic densities. The present study demonstrates that exposure of adult rats to both forms of silver leads to ultrastructural changes in synapses. However, it seems that small AgNPs lead to more severe synaptic degeneration, mainly in the hippocampal region of brain. The observations may indicate impairment of nerve function and, in the case of hippocampus, may predict impairment of cognitive processes.

Toxic effects of silver nanoparticles in mammals--does a risk of neurotoxicity exist?

Over the last decade, silver nanoparticles have become an important class of nanomaterials utilized in the development of new nanotechnologies. Despite the fact that nanosilver is used in many commercial applications, our knowledge about its associated risks is incomplete. Although a number of studies have been undertaken to better understand the impact of silver nanoparticles on the environment, aquatic organisms and cell lines, little is known about their side effects in mammalian organisms. This review summarizes relevant data and the current state of knowledge regarding toxicity of silver nanoparticles in mammals, as well as the accumulated evidence for potent neurotoxic effects. The influence of nanosilver on the central nervous system is significant because of evidence indicating that it accumulates in mammalian brain tissue.