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. 2022 Sep 16:15:992494.
doi: 10.3389/fnmol.2022.992494. eCollection 2022.

Ultrasonication effects on graphene composites in neural cell cultures

Łucja Dybowska-Sarapuk  1   2 , Weronika Sosnowicz  1   2 , Anna Grzeczkowicz  3 , Jakub Krzemiński  2 , Małgorzata Jakubowska  1   2
Affiliations

Ultrasonication effects on graphene composites in neural cell cultures

Łucja Dybowska-Sarapuk et al. Front Mol Neurosci. .

Abstract

Spinal cord injuries and neurodegenerative diseases, including Parkinson's, Alzheimer's, and traumatic brain injuries, remain challenging to treat. Nowadays, neural stem cell therapies excite high expectations within academia. The increasing demand for innovative solutions in regenerative medicine has drawn considerable attention to graphene materials. Due to unique properties, carbon materials are increasingly used as cellular scaffolds. They provide a biological microenvironment supporting cell adhesion and proliferation. The topography and mechanical properties of the graphene culture surface influence the forces exerted by the cells on their extracellular matrix. Which consequently affects the cell proliferation and differentiation. As a result, material properties such as stiffness, elasticity and mechanical strength play an important role in stem cells' growth and life. The ink unification process is crucial while the layer homogeneity is essential for obtaining suitable surface for specific cell growth. Different ink unification processes were tested to achieve appropriate layer homogeneity and resistivity to successfully applied the GNPs layers in neural cell electrostimulation. The GNP coatings were then used to electrostimulate mouse NE-4C neural stem cells. In this study, the authors investigated how the stimulation voltage amplitude's value affects cell behaviour, particularly the number of cells. Sinusoidal alternating current was used for stimulation. Three different values of stimulation voltage amplitude were investigated: 5, 10, and 15 V. It was noticed that a lower stimulation voltage amplitude had the most favourable effect on the stem cell count.

Keywords: cell electrostimulation; graphene nanoplatelets; graphene substrates; sonication method; spray-coating; surfactants.

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Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

FIGURE 1
FIGURE 1
Graphene ink fabrication scheme (Dybowska-Sarapuk et al., 2020).
FIGURE 2
FIGURE 2
Dependence of viscosity on shear rate for graphene inks with sonication times of 30, 60, 90, and 120 min using an ultrasonic bath and 1 min using a sonotrode.
FIGURE 3
FIGURE 3
Scanning electron microscope images showing microgeometry of coatings. (A,B) Microgeometry of substrates made with inks sonicated using sonotrode for 1 min, (C,D) with inks prepared using the ultrasonic bath for 30 min. (A) One minute – sonotrode, ×100 magnification; (B) 1 min – sonotrode, ×500 magnification; (C) 30 min – ultrasonic bath, ×100 magnification; (D) 30 min – ultrasonic bath, ×500 magnification.
FIGURE 4
FIGURE 4
Comparison of cell counts: seeded, cultured on polystyrene substrate and graphene substrates made with inks sonicated using the sonotrode and ultrasonic bath with and without electrostimulation (5, 10, and 15 V).
FIGURE 5
FIGURE 5
Comparison of cell counts cultured on graphene substrates and stimulated using different voltage amplitudes (5, 10, and 15 V).
FIGURE 6
FIGURE 6
Scanning electron microscope photograph of control group neural stem cells on graphene layer without stimulation.
FIGURE 7
FIGURE 7
Scanning electron microscope photographs showing neural stem cells on graphene layers stimulated with a voltage of 5 V amplitude: substrates made with sonicated inks using (A) sonotrode (B) ultrasonic bath.
See this image and copyright information in PMC

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