Assessment of functional changes in nanoparticle-exposed neuroendocrine cells with amperometry: exploring the generalizability of nanoparticle-vesicle matrix interactions

Abstract

Using two of the most commonly synthesized noble metal nanoparticle preparations, citrate-reduced Au and Ag, the impacts of short-term accidental nanoparticle exposure are examined in primary culture murine adrenal medullary chromaffin cells. Transmission electron microscopy (TEM), inductively coupled plasma atomic emission spectroscopy (ICP-AES) and Alamar Blue viability studies revealed that nanoparticles are taken up by cells but do not decrease cell viability within 48 hours of exposure. Carbon-fiber microelectrode amperometry (CFMA) examination of exocytosis in nanoparticle-exposed cells revealed that nanoparticle exposure does lead to decreased secretion of chemical messenger molecules, of up to 32.5% at 48 hours of Au exposure. The kinetics of intravesicular species liberation also slows after nanoparticle exposure, between 30 and 50% for Au and Ag, respectively. Repeated stimulation of exocytosis demonstrated that these effects persisted during subsequent stimulations, meaning that nanoparticles do not interfere directly with the vesicle recycling machinery but also that cellular function is unable to recover following vesicle content expulsion. By comparing these trends with parallel studies done using mast cells, it is clear that similar exocytosis perturbations occur across cell types following noble metal nanoparticle exposure, supporting a generalizable effect of nanoparticle-vesicle interactions.

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“…Carbon-fiber microelectrode amperometry examination of exocytosis in nanoparticle-exposed cells revealed that nanoparticle exposure lead to decreased secretion of chemical messenger molecules, of up to 32.5% at 48 h of gold nanoparticle exposure. Repeated stimulation of exocytosis demonstrated that these effects persisted during subsequent stimulations, meaning that nanoparticles do not interfere directly with the vesicle recycling machinery but also that cellular function is unable to recover following vesicle content expulsion [118]. Similar results were also observed in mast cells [118].…”

“…Repeated stimulation of exocytosis demonstrated that these effects persisted during subsequent stimulations, meaning that nanoparticles do not interfere directly with the vesicle recycling machinery but also that cellular function is unable to recover following vesicle content expulsion [118]. Similar results were also observed in mast cells [118].…”

“…An elegant approach to assessing nanoparticle impacts has been to look at the effect of citrate-reduced gold and silver nanoparticles on primary cultures of murine adrenal medullary chromaffin cells [118]. Carbon-fiber microelectrode amperometry examination of exocytosis in nanoparticle-exposed cells revealed that nanoparticle exposure lead to decreased secretion of chemical messenger molecules, of up to 32.5% at 48 h of gold nanoparticle exposure.…”

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“…(b) AFM is used to measure the adhesion force and stiffness of the cell membrane upon diesel NP exposure on human aortic endothelial cells [70]. (c) Carbon fiber microelectrode is a very sensitive technique with very high temporal resolution and has been used recently to identify the change in exocytosis behavior of endocrine or immune cells upon NP exposure [72][73][74].…”

“…The effect on cell viability when NP exposure was extended for 48 and 72 hours [74] was also studied. The effect of citrate reduced noble NPs, Au (28 nm) and Ag (61 nm), on neuroendocrine cells has been evaluated by Love and Haynes [72]. In this work, the uptake quantifications were measured using inductively coupled plasma-atomic emission spectroscopy (ICP-AES).…”