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  • Multicompartment Microreactors Prevent Excitotoxic Dysfunctions In Rat Primary Cortical Neurons.

Multicompartment Microreactors Prevent Excitotoxic Dysfunctions In Rat Primary Cortical Neurons.

Advanced biosystems (2020-09-02)
Adam Armada-Moreira, Joana E Coelho, Luísa V Lopes, Ana M Sebastião, Brigitte Städler, Sandra H Vaz
ABSTRACT

Excitotoxicity is a cellular phenomenon that comprises the consequences of toxic actions of excitatory neurotransmitters, such as glutamate. This process is usually related to overproduction of reactive oxygen species (ROS) and ammonia (NH4+ ) toxicity. Platinum nanoparticle (Pt-NP)-based microreactors able to degrade hydrogen peroxide (H2 O2 ) and NH4+ , are previously described as a novel therapeutical approach against excitotoxicity, conferring protection to neuroblasts. Now, it is demonstrated that these microreactors are compatible with rat primary cortical neurons, show high levels of neuronal membrane interaction, and are able to improve cell survival and neuronal activity when neurons are exposed to H2 O2 or NH4+ . Additionally, more complex microreactors are assembled, including enzyme-loaded liposomes containing glutamate dehydrogenase and glutathione reductase, in addition to Pt-NP. The in vitro activity of these microreactors is characterized and they are compared to the Pt-NP-based microreactors in terms of biological activity, concluding that they enhance cell viability similarly or more extensively than the latter. Extracellular electrophysiological recordings demonstrate that these microreactors rescue neuronal functionality lost upon incubation with H2 O2 or NH4+ . This study provides more evidence for the potential application of these microreactors in a biomedical context with more complex cellular environments.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
1,2-Dilinoleoyl-3-palmitoyl-rac-glycerol, ≥95% (TLC), liquid
Sigma-Aldrich
1,2-Dioleoyl-sn-glycero-3-phosphocholine, lyophilized powder
Sigma-Aldrich
Triton X-100, laboratory grade
Sigma-Aldrich
Ammonia Assay Kit, sufficient for 200 fluorometric tests