Evaluation of structural and biological characteristics of decellularized Wharton's jelly from human umbilical cord

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Introduction. Wharton's jelly of the human umbilical cord is a connective tissue of extraembryonic origin that maintains characteristics of the embryonic phenotype, including the capacity for rapid tissue regeneration and scar-free healing of fetal wounds. Decellularization refers to the removal of cells and cellular components from biological tissues while preserving the essential structural and compositional features of the extracellular matrix.

The aim of the work was to evaluate the structural and biological characteristics of the decellularized extracellular matrix from Wharton's jelly of the human umbilical cord.

Material and methods. Decellularization was performed by a detergent method using a sterile solution of sodium dodecyl sulfate at a concentration of 0.01% for 24 h at room temperature. The component composition of Wharton's jelly of the human umbilical cord before and after the decellularization process was assessed using spectral analysis methods. To study the biological characteristics of the decellularized extracellular matrix from Wharton's jelly of the human umbilical cord, the MTT test and the subcutaneous implantation model in mice were used.

Results. The content of total collagen by hydroxyproline in Wharton's jelly of the human umbilical cord before decellularization ranged from 244.8 to 507.2 μg/mg, and after – from 398.9 to 777.3 μg/mg, hyaluronic acid and sulfated glycosaminoglycans – from 11.5 to 16.5 and from 16.1 to 22.5 μg/mg before decellularization and from 15.6 to 22.1 and from 25.6 to 29.6 μg/mg after decellularization, respectively. Multiple collagen types (I, III, IV, V, VI, XII), as well as fibronectin, lumican, decorin, biglycan, and tenascin were identified in Wharton's jelly of the human umbilical cord. The absence of cytotoxicity of model media based on extracts from decellularized extracellular matrix from Wharton's jelly of the human umbilical cord was revealed. No signs of rejection or enhanced cellular inflammatory response were observed during subcutaneous implantation in mice.

Conclusions. The practical possibility of developing a drug and/or medical device for regenerative medicine based on decellularized extracellular matrix from Wharton's jelly of the human umbilical cord was demonstrated.

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作者简介

D. Tovpeko

S.M. Kirov Military Medical Academy

编辑信件的主要联系方式.
Email: tovpeko.dmitry@gmail.com
ORCID iD: 0000-0003-0286-3056
SPIN 代码: 3698-4656

Junior Researcher, Research Laboratory (Military Therapy) of the Research Department (Experimental Medicine) of the Research Center

俄罗斯联邦, Academician Lebedev st., 6, lit. G, Saint Petersburg, 194044

A. Kondratenko

S.M. Kirov Military Medical Academy; Saint Petersburg State Pediatric Medical University

Email: kondraa24@gmail.com
ORCID iD: 0000-0002-8511-5864
SPIN 代码: 1668-3497

Ph.D. (Biol.), Researcher, Research Laboratory (Cellular Technologies) of the Research Department (Medical and Biological Research) of the Research Center, Senior Lecturer, Department of Histology and Embryology named after Professor A.G. Knorre

俄罗斯联邦, Academician Lebedev st., 6, lit. G, Saint Petersburg, 194044; Litovskaya st., 2, Saint Petersburg, 194100

M. Okolitenko

S.M. Kirov Military Medical Academy

Email: matveyoko@mail.ru
ORCID iD: 0009-0002-4011-1699
SPIN 代码: 4319-3889

Cadet, Faculty 3 (Training of Doctors for the Aerospace Forces)

俄罗斯联邦, Academician Lebedev st., 6, lit. G, Saint Petersburg, 194044

D. Raguzina

S.M. Kirov Military Medical Academy

Email: daryavyacheslavovna@inbox.ru
ORCID iD: 0009-0008-0526-5557
SPIN 代码: 9154-8250

Junior Researcher, Research Laboratory (Water and Food Expertise) of the Research Department (Food and Water Supply) of the Research Center

俄罗斯联邦, Academician Lebedev st., 6, lit. G, Saint Petersburg, 194044

B. Zakopayko

S.M. Kirov Military Medical Academy

Email: bogdanzakopayko@gmail.com
ORCID iD: 0009-0004-9762-0886
SPIN 代码: 1589-8989

Junior Researcher, Research Laboratory (Military Therapy) of the Research Department (Experimental Medicine) of the Research Center

俄罗斯联邦, Academician Lebedev st., 6, lit. G, Saint Petersburg, 194044

A. Mittenberg

S.M. Kirov Military Medical Academy; Institute of Cytology

Email: a.mittenberg@gmail.com
ORCID iD: 0000-0002-3675-0597
SPIN 代码: 4791-0747

Ph.D. (Biol.), Senior Researcher, Research Laboratory (Cellular Technologies) of the Research Department (Medical and Biological Research) of the Research Center, Leading Researcher, Head of the Proteomics and Mass Spectrometry Group of the Center for Cellular Technologies

俄罗斯联邦, Academician Lebedev st., 6, lit. G, Saint Petersburg, 194044; Tikhoretsky pr., 4, Saint Petersburg, 194064

S. Shabelnikov

Institute of Cytology

Email: buddasvami@gmail.com
ORCID iD: 0000-0002-5693-5310
SPIN 代码: 1951-4801

Ph.D. (Biol.), Senior Researcher, Proteomics and Mass Spectrometry Group of the Center for Cellular Technologies

俄罗斯联邦, Tikhoretsky pr., 4, Saint Petersburg, 194064

E. Lapina

Institute of Cytology

Email: kathie.lap@yandex.ru

Senior Research Assistant, Cellular Biotechnology Laboratory of the Center for Cellular Technologies

俄罗斯联邦, Tikhoretsky pr., 4, Saint Petersburg, 194064

E. Alexander-Sinclair

Institute of Cytology

Email: elga.aleks@gmail.com
ORCID iD: 0000-0001-6704-3062
SPIN 代码: 2548-5932

Researcher, Cellular Biotechnology Laboratory of the Center for Cellular Technologies

俄罗斯联邦, Tikhoretsky pr., 4, Saint Petersburg, 194064

D. Zemlyanoy

Saint Petersburg State Pediatric Medical University

Email: zemlianoj@mail.ru
ORCID iD: 0000-0003-4716-809X
SPIN 代码: 3871-7531

Ph.D. (Med.), Associate Professor, Associate Professor of the Department of General Hygiene

俄罗斯联邦, Litovskaya st., 2, Saint Petersburg, 194100

E. Bagramyan

S.M. Kirov Military Medical Academy

Email: elin.bagramian@yandex.ru
ORCID iD: 0009-0009-8650-2426

Post-Graduate Student, Department of Obstetrics and Gynecology

俄罗斯联邦, Academician Lebedev st., 6, lit. G, Saint Petersburg, 194044

V. Chernov

S.M. Kirov Military Medical Academy; Agrophysical Research Institute

Email: vechernov@mail.ru
ORCID iD: 0000-0002-2440-3782
SPIN 代码: 8315-1161

Ph.D. (Biol.), Senior Researcher, Research Laboratory (Cellular Technologies) of the Research Department (Medical and Biological Research) of the Research Center, Senior Researcher, Department of Plant Light Physiology and Bioproductivity of Agroecosystems

俄罗斯联邦, Academician Lebedev st., 6, lit. G, Saint Petersburg, 194044; Grazhdanskiy pr., 14, Saint Petersburg, 195220

L. Kalyuzhnaya

S.M. Kirov Military Medical Academy

Email: terrestra@mail.ru
ORCID iD: 0000-0001-6698-4872
SPIN 代码: 1348-3306

Dr.Sc., (Med.), Senior Researcher, Research Laboratory (Tissue Engineering) of the Research Department (Medical and Biological Research) of the Research Center

俄罗斯联邦, Academician Lebedev st., 6, lit. G, Saint Petersburg, 194044

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2. Fig. 1. Scheme for obtaining decellularized extracellular matrix from Wharton's jelly of the human umbilical cord

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3. Fig. 2. Hydroxyproline content (left) and calculated total collagen content using a coefficient of 0.135 (right) in Wharton's jelly of the human umbilical cord before and after the decellularization process

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4. Fig. 3. Content of hyaluronic acid (left) and sulfated glycosaminoglycans (right) in Wharton's jelly of the human umbilical cord before and after the decellularization process

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5. Fig. 4. Proteomic profile of Wharton's jelly of the human umbilical cord before and after the decellularization process

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6. Fig. 5. Viability of human dermal fibroblasts cultured in model media based on extracts from decellularized extracellular matrix from Wharton's jelly of the human umbilical cord added immediately after adhesion (left) and 24 h after cell adhesion (right) (Tukey's test: *** – p <0.0001, ** – p <0.001, * – p <0.1, ns – not significant compared to control)

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7. Fig. 6. Morphology of human dermal fibroblasts cultured in model media based on extracts decellularized extracellular matrix from Wharton's jelly of the human umbilical cord added immediately after adhesion (left) and 24 h after cell adhesion (right)

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8. Fig. 7. Decellularized extracellular matrix from Wharton's jelly of the human umbilical cord under mouse skin at different days after implantation. Hematoxylin and eosin staining

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