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Galvanotaxis of fibroblast cells cultured on carbon nanotubes films


I.I. Bobrinetskiy, R.A. Morozov, A.S. Seleznev, R.Ya. Podchernyaeva, G.R. Mikhailova, O.A. Lopatina, I.A. Suetina

The results of human embryonic fibroblasts cell cultured in a pulsed electric field on the coverslips with the conductive films consisting carbon nanotubes and bovine serum albumin composite were presented. Single-walled carbon nanotubes (SWNT) and bovine serum albumin (BSA) composite, was formed under the action of laser radiation. Conductive nanocomposite film on coverslips without and with cell culture were studied using atomic force microscopy and optical microscopy. The galvanotaxis of cells when applying pulse voltages from 10 to 5000 mV was investigated. For the implementation of electrical stimulation in the growth of cells was modified by 6-well plate. As the electrodes were used acupuncture needles Doriva of biocompatible alloy 40KHNM-VI with a gold coating. The experiment used a cell line EPP-T (laboratory tissue cultures FGBU "Research Institute of Virology. Ivanovsky" Health Ministry of Russia), the tissue was obtained from a line of human embryonic fibroblasts (skin and muscle tissue). To assess the proliferative activity of cells on SWNT was used MTT test.Studies have shown that if you directly connect voltage after planting cells are poorly fixed on the surface. Particularly significant was the effect for potentials above 100 mV. In general, the current is increased, there is accumulation of proliferating cells in areas of high film thickness of nanotubes and their characteristic orientation along the electric field lines at voltages above 100 mV (10 nA currents). In small fields of short electric pulses lead to an increase in rate of proliferation of CP by 20%. It remains unclear, the effect of inhibition of cell growth at voltages above 100 mV. For large values of voltage cells begin to come into direct contact with each other and form the structure of the properties close to the tissue. Thus, the electrically conductive substrate was designed based on the SWNT and BSA, and can be used for culturing cells with the electrostimulation and tissue regeneration.
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