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Chemically-functionalized single-walled carbon nanotubes increase the expression of glial fibrillary acidic protein in the mouse primary astrocytes exposed to severe in vitro traumatic brain injury

Astrocytes are central nervous system (CNS) cells with an important role in neuronal regeneration following brain injury. The aim of this study was to investigate the effects of single-walled carbon nanotubes (SWCNTs), chemically functionalized with poly-m-aminobenzene sulfonic acids (PABS), on the oxidative protein damage and the expression of glial fibrillary acidic protein (GFAP) and the inducible nitric oxide synthase (iNOS) in astrocytes exposed to an vitro model of severe traumatic brain injury (TBI). Primary mouse astrocytes were grown on 6-well plates with deformable membranes and subjected to stretch injury, mimicking severe TBI. PABS-SWCNTs were applied to severely injured cells at 1 h post-injury. Cells were collected at 24 h following stretch and the lysates prepared for the analyses. Dot blot method was used to detect the levels of oxidatively damaged protein. Western blot analyses were used to determine the expressions of GFAP and iNOS. Non-treated cells were used as the control. Dot blot assay demonstrated that the stretch injury and the treatment with PABS- SWCNTs did not cause significant change in the levels of oxidatively damaged proteins. We observed no change in the expression of the inflammatory marker iNOS. Conversely, application of PABS- SWCNTs significantly elevated the expression level of GFAP compared to the results obtained in the samples of both control cells and the non-treated, severely stretched astrocytes. Our preliminary results suggest that the application of the investigated nanomaterial, applied as colloidal solute, to the primary astrocytes exposed to severe in vitro TBI, causes increase in the GFAP levels. This is a finding with possible relevance related to the role of GFAP in the trafficking of glutamate transporters which could be associated to the uptake of extracellular glutamate and maintenance of homeostasis in the CNS. This research was fully supported by the Croatian Science Foundation grant UIP-2017-05-9517 to KP.

astrocytes ; glial fibrillary acidic protein ; inducible nitric oxide synthase ; mouse ; nanotubes, carbon ; oxidative stress ; brain injury, traumatic

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