Widefield Fluorescence Microscopy in Preclinical Studies of Biomedical Materials, Scaffolds, and Biomedical Cell Product
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MER MATERIALS FOR BIOMEDICINE
Widefield Fluorescence Microscopy in Preclinical Studies of Biomedical Materials, Scaffolds, and Biomedical Cell Products M. N. Egorikhinaa,*, D. Ya. Aleinika, I. N. Charykovaa, Yu. P. Rubtsovaa, V. V. Yudinb, and A. G. Morozovb a Privolzhsky
b
Research Medical University, Nizhny Novgorod, 603005 Russia Razuvaev Institute of Organometallic Chemistry, Russian Academy of Sciences, Nizhny Novgorod, 603137 Russia *e-mail: [email protected] Received January 16, 2020; revised February 25, 2020; accepted February 25, 2020
Abstract—We present the results of studies of the interaction of cells with various materials and products from them intended for biomedical use. Widefield fluorescence microscopy enables evaluating the cytotoxicity of samples, cell viability, morphology, and proliferative activity in the process of interaction with the test samples, as well as adhesion, migration, and cell distribution both on the surface and in the structure of scaffolds and biomedical cell products. The methods of fluorescence microscopy make possible intravital studies of the interaction of cells with materials without additional processing or destruction of samples. DOI: 10.1134/S1063784220090133
INTRODUCTION The development of regenerative medicine is inextricably linked with the search for new materials. Innovative materials are used independently as medical devices or serve as the basis for scaffolds and biomedical cell products (BMCPs). Materials intended for biomedical use are extremely diverse in terms of characteristics, composition, and properties: metals, bioceramics, curable synthetic polymers, decellularized matrices, hydrogels (synthetic/natural), etc. Materials play a key role in regeneration processes, ensuring successful cellular events and integration of structures into the patient’s body. For this reason, their interaction with cells at the stage of development of materials should be evaluated. There is no doubt that, if the material adversely affects the morphology and functional activity of cells in an in vitro model, it will have an adverse effect when applied in clinical practice. Preclinical studies in an in vitro model are limited to the study of the cytotoxicity of the material, as a rule, by indirect methods—for example, using the widespread MTT test. Visual in vivo assessment of the state and functioning of cells on materials using standard methods of light and phase-contrast microscopy is difficult, since most often, these objects are not transparent. Therefore, a more detailed study, such as the characteristics of cell adhesion, their distribution on the surface and/or in the structure of the material, and the viability and change in proliferative activity in the process of interaction with materials, requires more complex research methods [1] related to the death of cells and/or their separation from the matrix
and its destruction. Additional manipulations can lead to cell loss and, accordingly, significant errors in the analysis of the results. All of the above
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