THE INFLUENCE OF MESENCHYMAL STEM CELLS ON THE COURSE OF THE IMMUNE PROCESS IN RATS WITH ACUTE PERIODONTITIS
DOI:
https://doi.org/10.11603/mcch.2410-681X.2021.i2.12259Keywords:
acute periodontitis, inflammatory process, immunological reactivity, mesenchymal stem cellsAbstract
Introduction. Methods of regenerative therapy are beginning to be widely used in medicine. Treatment of periodontitis is a complex task of dentistry, therefore, there is a search for new methods that would be effective in this pathology.
The aim of the study – to learn the effect of stem cells on the state of cellular and humoral immunity in conditions of experimental periodontitis.
Research Methods. The study was carried out on white out bred male rats weighing 180–200 g. Periodontitis was caused by the introduction of lipopolysaccharide into the gum tissue at 40 microliters (1 mg/ml) every other day for 14 days. On days 1, 7, 14, and 21 after the last LPS injection, the rats were decapitated under thiopental anesthesia (50 mg/kg). Material from intact animals served as control. MSCs were obtained from pregnant females, approximately on the 21–24th day of pregnancy. An enzymatic method was used to obtain viable MSCs. The cultivation was carried out in a CO2 incubator at a temperature of 37 °C and a CO2 concentration of 5 %. Stem cells were injected into the gum of rats with a single injection at the rate of 1 million cells per 1 kg of body weight. To maximize the preservation of cell viability, MSCs were injected within 30 min after the suspension was obtained. Studies of the cellular (CD4 +, CD8 +, CD3 +, CD20 +) immune system were performed by immunofluorescence method using monoclonal antibodies to CD4 + -, CD8 + -, CD3 + -and CD20 + -antigens of rats conjugated to fluorescein isothiocyanate (FITC) produced by “Beckman Coulter” (USA). Immunoreactivity of the organism was studied by the content of serum immunoglobulins A, M, G by the method of solid-phase enzyme-linked immunosorbent assay using a set of reagents "eBioscience, Inc" using the analyzer "StatFax". The obtained digital data were processed by the method of variation statistics. The significance of differences in the compared parameters between different samples was determined by using the Student's t-test (with a normal distribution of results) or Mann-Whitney (in the case of a distribution that was not normal).
Results and Discussion. Under the conditions of modeling of acute periodontitis, the production of serum immunoglobulins increased significantly in the initial stages of the experiment with a subsequent decrease to the 21st day. Under the conditions of MSC injection, the production of immunoglobulins was less intense than in animals without correction. The level of CD4 + cells in animals with acute periodontitis increased significantly in the initial stages, with a subsequent decrease to the 21st day. After the injection of MSC, the level of CD4 + cells was lower and on the 1st day was 130.8 % of healthy animals, which was 88.6 % of the level of animals that were not corrected. By the day 21, CD4 + cell levels continued to decline and did not differ significantly from animals without pathology. The dynamics of CD8 + cell levels were similar, but the growth was less than that of CD4 +. The dynamics of the CD4 + / CD8 + regulatory index indicates a significant increase in animals with acute periodontitis, with a further decrease to the 21st day The content of CD3 + and CD20 + cells in the serum of animals with acute periodontitis on the 1st day significantly exceeded that of intact animals with a further decrease to the 21st day. The correction with the use of MSCs was accompanied by a less intense increase in the level of CD3 + and CD20 + cells.
Conclusions: In animals with acute periodontitis there is a significant increase in CD4 + lymphocytes, a violation of the ratio of major subpopulations of lymphocytes (CD4 + and CD8 +), accompanied by a significant increase in the immunoregulatory index, and dysfunction of the humoral immune system, manifested by blood imbalance and reducing the resistance of the humoral part of the immune system. The use of MSCs significantly compensates for the imbalance caused by the pathological process of the immune system, contributing to the normalization of the immunoregulatory index and the main classes of immunoglobulins.
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