CYTOKINE CONTENT IN CELL CULTURE SUPERNATANTS WHEN CULTURED WITH VARIOUS FORMS OF PLATELET LYSATE



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Abstract

The literature describes the high content of a wide range of cytokines and growth factors in platelet lysate (PL). Furthermore, it is noted that its origin from donor blood is more suitable for culturing human cells. We have developed a trophic-proliferative supplement based on PL in both native and lyophilized forms. The effects of this trophic-proliferative supplement are not limited to its influence on the proliferative activity of stem cells. The positive results of PL in the treatment of wounds of various origins are well known. These results are mediated not only by the recruitment of activated neutrophils and macrophages to the wound in the early stages of healing, but also by immunoregulatory effects during the regenerative stage of healing. It has been noted that the most positive results are achieved with a combination of platelet lysate products and fibrin-based preparations. The aim of our study was to analyze cytokine levels in MMSC cell culture supernatants obtained with the addition of defibrinated inactivated human serum or phosphate-buffered saline, enriched with platelet lysate and fibrin. Methods. We assessed the levels of heparin-binding factor (FGF), platelet-derived growth factor (PDGF), vascular endothelial growth factor (VEGF), and the proinflammatory mediators tumor necrosis factor-alpha (TNF-α) and interleukin 1-beta (IL-1β). Results. The levels of the studied growth factors (PDGF, FGF, VEGF), and the pleiotropic cytokine IL-1β) were significantly higher than plasma reference values ​​in all samples, regardless of lyophilization. However, different growth supplement compositions demonstrated different levels of growth factor secretion. TNF-α levels in all study groups were significantly lower than the device's sensitivity and reference values ​​in blood plasma. Conclusions. The experimental results demonstrate a theoretical basis for using 10% fibrin-enriched platelet lysate for wound treatment. We hypothesize that mesenchymal stem cells present in tissues, under the influence of the growth supplement we developed, can actively secrete a number of growth factors (PDGF, FGF, VEGF, etc.), which stimulate angiogenesis, the proliferation of epithelial and muscle cells, fibroblasts, and extracellular matrix synthesis, promoting wound healing. The observed secretion of IL-1β differed little between the study groups; however, its presence provides an immunomodulatory effect and activates connective tissue cells. 

About the authors

Andrey Goncharov

Federal State Autonomous Educational Institution of Higher Education "Immanuel Kant Baltic Federal University" of the Ministry of Science and Higher Education of the Russian Federation, Kaliningrad, Russia

Email: agoncharov59@mail.ru
ORCID iD: 0000-0001-6967-8838
SPIN-code: 7159-0402
Scopus Author ID: 57196504924
ResearcherId: D-3170-2018

MD, PhD, Senior Researcher, Center for Immunology and Cellular Biotechnology, Immanuel Kant Baltic Federal University; Ministry of Education and Science of the Russian Federation

Russian Federation, RF, Kaliningrad, st. A. Nevsky, 14

Valeria Shupletsova

Federal State Autonomous Educational Institution of Higher Education "Immanuel Kant Baltic Federal University" of the Ministry of Science and Higher Education of the Russian Federation, Kaliningrad, Russia

Email: vshupletsova@mail.ru
ORCID iD: 0000-0001-7243-9731
SPIN-code: 5736-4492

Senior Researcher at the Center for Immunology and Cellular Biotechnology

Russian Federation, RF, Kaliningrad, st. A. Nevsky, 14

Olga Melashchenko

Federal State Autonomous Educational Institution of Higher Education "Immanuel Kant Baltic Federal University" of the Ministry of Science and Higher Education of the Russian Federation, Kaliningrad, Russia

Email: OMelashchenko@kantiana.ru
ORCID iD: 0000-0002-0892-1168
SPIN-code: 2600-9368

research fellow at the Center for Immunology and Cellular Biotechnology

Russian Federation, RF, Kaliningrad, st. A. Nevsky, 14

Natalya Gazatova

Federal State Autonomous Educational Institution of Higher Education "Immanuel Kant Baltic Federal University" of the Ministry of Science and Higher Education of the Russian Federation, Kaliningrad, Russia

Email: n_gazatova@mail.ru
SPIN-code: 1174-7330

Senior Researcher, Head of the Laboratory of Experimental Studies of Blood Products at the Center for Immunology and Cellular Biotechnology

Russian Federation, RF, Kaliningrad, st. A. Nevsky, 14

Olga Khaziakhmatova

Federal State Autonomous Educational Institution of Higher Education "Immanuel Kant Baltic Federal University" of the Ministry of Science and Higher Education of the Russian Federation, Kaliningrad, Russia

Email: hazik36@mail.ru
ORCID iD: 0000-0002-5525-3529
SPIN-code: 5906-9527

PhD (Biology), Senior Researcher, Center for Immunology and Cellular Biotechnology

Russian Federation, RF, Kaliningrad, st. A. Nevsky, 14

Vladimir Malashchenko

Federal State Autonomous Educational Institution of Higher Education "Immanuel Kant Baltic Federal University" of the Ministry of Science and Higher Education of the Russian Federation, Kaliningrad, Russia

Email: Vlmalashchenko@kantiana.ru
ORCID iD: 0000-0003-3437-0926
SPIN-code: 9195-4537

PhD (Biology), Senior Researcher, Center for Immunology and Cellular Biotechnology

Russian Federation, RF, Kaliningrad, st. A. Nevsky, 14

Nina Kabanchuk

GBUZ "Blood Transfusion Station of the Kaliningrad Region", Ministry of Health of the Kaliningrad Region, Kaliningrad, Russia

Email: spk@infomed39.ru

Acting Chief Physician, State Healthcare Institution "Blood Transfusion Station of the Kaliningrad Region"

Russian Federation, RF, Kaliningrad, st. Chkalova, 29

Irina Zatrusckina

GBUZ "Blood Transfusion Station of the Kaliningrad Region", Ministry of Health of the Kaliningrad Region, Kaliningrad, Russia

Email: zatrusckina@mail.ru
ORCID iD: 0009-0009-8836-3925

Nurse, State Healthcare Institution "Blood Transfusion Station of the Kaliningrad Region"

Russian Federation, RF, Kaliningrad, st. Chkalova, 29

Larisa Litvinova

Federal State Autonomous Educational Institution of Higher Education "Immanuel Kant Baltic Federal University" of the Ministry of Science and Higher Education of the Russian Federation, Kaliningrad, Russia

Author for correspondence.
Email: larisalitvinova@yandex.ru
ORCID iD: 0000-0001-5231-6910

MD, PhD, Associate Professor, Director of the Center for Immunology and Cellular Biotechnology, Immanuel Kant Baltic Federal University; Ministry of Education and Science of the Russian Federation

Russian Federation, RF, Kaliningrad, st. A. Nevsky, 14

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Copyright (c) Goncharov A., Shupletsova V., Melashchenko O., Gazatova N., Khaziakhmatova O., Malashchenko V., Kabanchuk N., Zatrusckina I., Litvinova L.

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