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No1-2(5) 2022

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DOI 10.37219/2528-8253-2022-1-29
Minin YuV, Karas AF, Karas GA, Kucherenko TI, Chaika SP, Virych PA, Shuvalova NS, Raskaliei TY
Morphological investigation of reparative processes in the area of the defect of cartilage tissue of the external ear of the rabbit after the local application of allogenic mesenchymal stem cells
Minin Yuriy V.
State Institution “O.S. Kolomiychenko Institute of Otolaryngology of National Academy of Medical Science of Ukraine”; Kyiv, Ukraine.
Department of inflammatory diseases of ENT-organs, leading researcher.
Doctor of Medical Sciences, Professor
ORCID ID: 0000-0002-7836-3480
Scopus Author ID: 6508004691
Email: mininy1956@gmail.com
ORCID ID: https://orcid.org/0000-0002-7836-3480
Scopus Author ID: 6508004691

Karas Anton F.
State Institution “O.S. Kolomiychenko Institute of Otolaryngology of
National Academy of Medical Science of Ukraine”; Kyiv, Ukraine
Laboratory of Biophysics
Head of Laboratory, PhD of Biological Sciences
E-mail: akaras2022@gmail.com
ORCID ID: https://orcid.org/0000-0003-2324-780X

Karas Halyna A.
State Institution “O.S. Kolomiychenko Institute of Otolaryngology of
National Academy of Medical Science of Ukraine”; Kyiv, Ukraine
Laboratory of Biophysics,
Leading researcher, PhD of Biological Sciences
E-mail: gkaras@ukr.net
ORCID ID: https://orcid.org/0000-0003-3098-3813

Kucherenko Tetiana I.
State Institution “O.S. Kolomiychenko Institute of Otolaryngology of National Academy of Medical Science of Ukraine”; Kyiv, Ukraine.
Department of inflammatory diseases of ENT-organs, Senior Researcher
E-mail: tkucherenko@hotmail.com
ORCID ID: https://orcid.org/0000-0002-2158-8101
Scopus Author ID: 6701383048

Chaika Svitlana P
State Institution “O.S. Kolomiychenko Institute of Otolaryngology of National Academy of Medical Science of Ukraine”; Kyiv, Ukraine
Laboratory of Biophysics
Senior researcher
Candidate of Biological Sciences
E-mail: svetlach555@gmail.com
ORCID ID: https://orcid.org/0000-0003-4324-5529

Virych Pavlo A.
State Institution “O.S. Kolomiychenko Institute of Otolaryngology of National Academy of Medical Science of Ukraine”; Kyiv, Ukraine
Laboratory of Biophysics
Researcher, PhD of Biological Sciences
E-mail: sphaenodon@ukr.net
ORCID ID: https://orcid.org/0000-0002-1463-1992
Scopus Author ID: 57193401578

Shuvalova Nagiya S
State Institution “O.S. Kolomiychenko Institute of Otolaryngology of
National Academy of Medical Science of Ukraine”; Kyiv, Ukraine
Laboratory of Biophysics
Senior Research Fellow, PhD of Medical Sciences
E-mail: Shuvalovanadiia@gmail.com
ORCID ID: https://orcid.org/0000-0002-6390-5996

Raskaliei Tetiana Ya.
Bogomolets National Medical University
Kafedra histolohiyi ta embriolohiyi Dotsent Kandydat medychnykh nauk
Department of Histology and Embryology
Assistant professor, PhD of Medical Sciences
E-mail: t_darmogray@ukr.net
ORCID ID: https://orcid.org/0000-0002-6737-2381
Scopus Author ID: https://www.scopus.com/authid/detail.uri?authorId=57195438833

Abstract

Purpose: to study the features of the restoration of rabbits outer ear elastic cartilage tissue in the area of the defect after its filling with allogeneic mesenchymal stem cells (MSCs).
Materials and methods: Studies were performed on 24 sexually mature rabbits. All animals under general anaesthesia underwent surgery to create a standardized linear defect and were divided into experimental (15 rabbits) and control (9 rabbits) groups. In the experimental group, the defect was filled with suspensions of MSCs in the amount of 5 million cells, and in the control group of cells was not introduced. MSCs were obtained from the
umbilical cord of rabbits by the enzymatic method. After the first passage of the MSCs culture, was studied MSCs stem potential and tested ability to differentiating. Material for morphological studies was taken after 1, 2 and 3 months, cryostat sections were prepared hematoxylin-eosin and picrofuxin staining were performed according to van Gizon, as well as morphometry.
Results: The introduction of suspensions of allogeneic MSCs in the experiment in the cartilage defect of  the outer ear of rabbits causes healing with the activation of pre-existing embryonic cartilage cells, the appearance of cells in a state of division in the adjacent zone, activation of angiogenesis and promotes fibrotization with a predominance of loose connective tissue 1 month after surgery. After 3 months, a more pronounced activation of chondrogenesis is observed with the growth of chondrocytes and their increased number in lacunae, the appearance of isogenic groups of cells, which is combined with vascular activation and the growth of dense scar connective tissue,increased fibroblasts and the number of collagen fibers in the defect area, as well as in the absence of a complete restoration of the characteristic cartilage structure. At that time, in the control group, in the process of defect healing 1 month after the operation, along with the detection of a reduced zone of activation of cartilage chondrocytes and collagen fibers, the development of destructive-dystrophic processes of chondrocytes and the extracellular cartilage matrix was observed. The morphometry data quantitatively confirm a more pronounced activation of chondrogenesis in the areas adjacent to the cartilage defect under the conditions of MSC injection compared to the control. The difference is 20.8% 1 month after the operation and 14.2% 3 months later. The results obtained indicate the promise of using MSCs to activate reparative processes in the area of rabbits outer ear cartilage tissue defect.

Keywords

reparative medicine, cartilage, mesenchymal stem cells, experimental studies.


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