I.R. Ishmukhametov*, E.V. Rozhina**, F.S. Akhatova***, V.G. Evtugyn****, A.O. Rozhin*****, R.F. Fakhrullin******
Kazan Federal University, Kazan, 420008 Russia
E-mail: *irishmukhametov@gmail.com, **rozhinaelvira@gmail.com, ***akhatovaf@gmail.com, ****hitachiht7700@gmail.com, *****rozhinartemkzn@gmail.com, ******kazanbio@gmail.com
Received February 11, 2020
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DOI: 10.26907/2542-064X.2020.4.557-572
For citation: Ishmukhametov I.R., Rozhina E.V., Akhatova F.S., Evtugyn V.G., Rozhin A.O., Fakhrullin R.F. Preparation and description of magnetic modified colloidal particles of silicon dioxide for recognition of HeLa cells. Uchenye Zapiski Kazanskogo Universiteta. Seriya Estestvennye Nauki, 2020, vol. 162, no. 4, pp. 557–572. doi: 10.26907/2542-064X.2020.4.557-572. (In Russian)
Abstract
Modification of the cell surface by the methods of nanoarchitectonics allows changing the physical and chemical properties of cells. Thus, it is possible to get imprinted colloid particles based on template cells that are able to recognize and selectively attach to cells. Application of nanomaterials in the inorganic coating composition expands their biomedical potential. For example, doping of cell imprints with magnetic nanoparticles allows manipulating the cells associated with shell fragments by the external magnetic field. In this work, we developed colloidal cell imprints based on silicon dioxide doped with iron oxide magnetic nanoparticles capable of binding to HeLa cells. The method of chemical coprecipitation was used to synthesize iron oxide magnetic nanoparticles. The hydrodynamic size and ζ-potential of nanoparticles were measured by the dynamic light scattering method. The morphology of magnetic nanoparticles was analyzed with the help of transmission electron microscopy and dark-field microscopy. The sol-gel process was used to coat HeLa cells by silicic acid derivatives doped with magnetic nanoparticles. Coating formation on cells was observed with scanning electron microscopy. Colloid cell imprints were obtained after the breakage of the inorganic coating by the ultrasonic treatment. Subsequently, colloid particles were cultivated with the HeLa cell line and observed with bright-field microscopy. Additionally, the morphology of the coating and cell imprints was visualized by the atomic force microscopy. Spherical magnetic nanoparticles with a diameter of about 110 nm were obtained. Silica coating formation on the HeLa cells was demonstrated. Furthermore, it was established that colloid imprints obtained after the decomposition of the silica-based shell are capable of binding to cells. Therefore, we successfully manipulated the cells coated with the silica-based shell doped with magnetic nanoparticles.
Keywords: nanoarchitectonics, magnetic nanoparticles, cell surface engineering, HeLa cell line, cell encapsulation
Acknowledgments. The work is performed according to the Russian Government Program of Competitive Growth of Kazan Federal University, as well as supported by the Russian Foundation for Basic Research and the Government of the Republic of Tatarstan (project no. 18-44-160001), by the Russian Foundation for Basic Research (project no. 18-34-00306), and by the grant for young scientists of the Republic of Tatarstan (project no. 05-129-sh G/2020).
Some microscopic images were obtained using the equipment of the Electron Microscopy Laboratory, Interdisciplinary Center for Analytical Microscopy, Kazan Federal University. Scanning electron microscopic images were obtained as part of the program of short-term scientific and educational internships in electron microscopy of carbon-based materials at N.D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences (operator: I.V. Chistyakov).
Figure Captions
Fig. 1. Formation of an inorganic coating on the mammalian cells and subsequent fragmentation of this coating to recognize template cells.
Fig. 2. Dark-field image of colloidal magnetic nanoparticles coated with polyallylamine hydrochloride (a), spectral profile of the individual magnetic particle (b). Scanning electron microscopic image of the magnetic nanoparticles coated with polyallylamine hydrochloride (c). Hydrodynamic size (d) and ζ-potential (e) of the magnetic nanoparticles coated with polyallylamine hydrochloride.
Fig. 3. Effect of the external magnetic field on the shifting of HeLa cells with the SiO2-based coating and magnetic nanoparticles towards the magnet (a) and the movement pattern of the encapsulated cells in the external magnetic field (b). Circles – cells with the tracked movement. Arrows – shifting of cells at certain points in time.
Fig. 4. Scanning electron microscopic images of the eukaryotic HeLa cells with the SiO2 coating (a) and with the SiO2@MNP-PAH coating (b); HeLa cells bound with the shell fragments based on SiO2 (c) and SiO2@MNP-PAH (d); HeLa cells selectively bound with the fragments in the suspension of E. coli cells (e). Red arrows – magnetic nanoparticles, white circles – fragments on the cells. Cell nuclei stained with DAPI.
Fig. 5. AFM visualization of the HeLa cell line: control cells (a–c); cells with a silica coating doped with magnetic nanoparticles (d–f); adhesion of the shell fragments on the cell surface (g–i). a, d, g – surface topography in the Peak Force Error channel; c, f, i – adhesion of the surface in the Adhesion channel. White arrows – magnetic nanoparticles, yellow arrows – adherent shell fragments.
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