BLOOD CYTOKINE PROFILE AND LESION SITE REPAIR IN DYNAMICS OF EXPERIMENTAL THERMAL TRAUMA AFTER LOCAL AND SYSTEMIC MELATONIN ADMINISTRATION

Burn injuries are one of the key medical and social problems. Despite the significant achievements in combustiology, the slow healing and the appearance of infection are the key problems in burn patients, which lead to a longer hospitalization period, to reduction of life quality and to emotional disorders. Up to 70% of all complications after thermal trauma (TT) are connected with infection – first of all, pneumonia, infections of urinal tract. The forming of infectious complications, including sepsis, after TT is associated with excessive immunosuppressive reactions, as compensation for a long, stable proinflammatory response, in particular, owing to hyperproduction and effects of IL-10, IL-4, TGF-β. Aim: to study the influence of systemic and local usage of MT with original dermal film (DF) on reparation and serum cytokine concentration indicators in dynamics of experimental TT. The study was conducted using 84 rats – males of Wistar line, which were divided randomly into 4 groups: 1st group (n = 12) – intact monitoring, 2nd group (n = 30) – animals with TT, 3rd group (n = 21) – animals with TT and DF with MT use on the region of burn, 4th group (n = 21) – animals with TT and intraperitoneal injection of MT. To model TT of IIIA degree and relative area 3,5%, isolated skin area of interscapular area was immersed in distilled water at a temperature of 98-99 °С at 12 s. The DF with MT (at a concentration of 0.005 g/g) on 12 sm2 – area in 3rd group was used daily for 5 days. The MT was injected intraperitoneally daily at the dose of 10 mg/kg for 5 days. The wound area was calculated, the interleukin-4 (IL-4), tumor necrosis factor alpha (TNFα), interferon-gamma (IFNγ) were determined in serum on 5th,10th and 20th day from the moment of TT induction in each group. During experimental TT in dynamic monitoring from 5th to 20th day the absolute and relative areas of wound defect are reduced, because of that the epithelization speed and its part of area reduction are progressively increasing, on 5th,10th and 20th day the concentration of TNFα and IL-4 in serum is increasing with maximum values on 10th day of monitoring. Local usage of MT in DF during TT accelerates the healing of burn wound and lowers the TNFα and IL-4 concentration in serum on 5th, 10th and 20th day. Intraperitoneal use of MT during TT accelerates the healing of burn wound and lowers the TNFα and IL-4 concentration in serum on 5th and 20th day. The reparation accelerating effect of MT during TT is more expressed in locale usage in DF rather than using intraperitoneal injection

Immune changes at the injured sites in oxazolone-induced ulcerous colitis: Influence of ozone therapy

Impaired immunoregulation and development of autoimmune response to antigens of own intestinal microbiota and inflammation-altered antigens of colonic cells represent the key links in pathogenesis of inflammatory bowel diseases. Multimodal biological effects of ozone presunme the usage of local and systemic ozone therapy in complex treatment of many inflammatory diseases of the gastrointestinal tract. The aim of our work was to study effects of intraperitoneal and rectal ozone therapy upon immune parameters of the lesion focus in oxazolone-induced ulcerative colitis in the course of time. The study was carried out on 64 adult male inbred Wistar rats weighing 240±20 g. Experimental ulcerative colitis was produced by oxazolone treatment (4-ethoxymethylene-2-phenyl-2-oxazolin-5-one) (SigmaAldrich, USA). The ozone-oxygen mixture was injected intraperitoneally or rectally at a concentration of 1.0-1.2 mg/l, once a day, in a volume of 10 ml, at the 6-day course. The results of experiments were recorded on the days +2, +4 and +6. The concentrations of IL-17 and IL-23 was determined in a homogenate of intestinal tissues (Bender Medsystems, Austria) using a Personal LAB analyzer; expression of CD4 and FoxP3 on intestinal lymphocytes was determined by immunohistochemistry technique (ElisaKit, China). The observed tissue damage of large intestine showed an increase from day 2 to day 6 of oxazolone-induced ulcerative colitis. The total number of lymphocytes significantly increased upon development of experimental colitis, with parallel decrease in the number of CD4+ lymphocytes and FoxP3-positive T lymphocytes. IL-17 and IL-23 concentrations in the tissues increased with the severity of inflammatory changes in the lesion focus. Intraperitoneal ozone administration was associated with significant reduction of lymphocyte contents in the damaged tissues on the 6th day, whereas the numbers of CD4+ and FoxP3 positive T lymphocytes normalized by the 6th day. The levels of IL-17 and IL-23 increased from day 2 to day 6, with a lower IL-23 values on day 6 as compared with non-treated animals. Rectal administration of ACS led to the normalization of FoxP3 cells on the 6th day to the values of intact animals. The levels of proinflammatory cytokines (IL-17 and IL-23) significantly decreased on the 6th day as compared to the group of animals without treatment, which could be due to anti-inflammatory properties of ozone

Clinical and Morphological Analysis of Efficacy of Intraperitoneal Ozone Application in Experimental Colitis: Preclinical Randomized Experimental Study

Background. Inflammatory bowel diseases — Crohn’s disease and ulcerative colitis — are chronic gastrointestinal diseases affecting young people of working age. An alternative to basic therapy (5-aminosalicylic acid) for inflammatory bowel disease is the use of ozone, which has anti-inflammatory, immunomodulatory, antibacterial properties and no side effects in therapeutic concentrations. Objective. To perform clinical and morphological analysis of efficacy of intraperitoneal ozone application in experimental colitis.Methods. The study was conducted on 73 male Wistar rats weighing 200-250 g. The animals were divided into four groups by simple randomization. Check studies were performed on the second, fourth and sixth days. Oxazolone-induced colitis was simulated in two stages using a 3%-alcohol oxazolone solution. Ozone-acid mixture was obtained on “UOTA-60-01” unit (“Medozone”, Russia). Rectal suppositories with 5-aminosalicylic acid were prepared on the basis of rectal suppositories “Salofalk”. Clinical status was assessed daily according to the disease activity index (DAI) scale. Morphological evaluation of colon lesion tissue fragments was carried out using a PrimoStar microscope (CarlZeiss, Germany). Colon tissue damage was assessed using tissue damage index (TDI). Statistical analysis was conducted with SPSS Statistics 19 (IBM, USA).Results. Clinical and morphological picture of the large intestine lesion in oxazolone-induced colitis on days 2, 4 and 6 is consistent with the changes typical of inflammatory bowel disease in humans. Daily intraperitoneal insufflation of ozone at a dose of 0.05 mg/kg in oxazolone-induced colitis leads to partial restoration of DAI, reduction in neutrophils, eosinophils, histiocytes, and fibroblasts in the lesion, as well as to a decrease in ulcerous defect diameter and TDI. The effects of intraperitoneal insufflations of ozone in oxazolone-induced colitis as compared to rectal suppositories with 50 mg of 5-aminosalicylic acid every 12 hours were less marked for the DAI index on day 4; for the number of eosinophils, plasma cells, histiocytes — on day 2, 4 and 6; for lymphocytes — on day 6.Conclusion. Clinical and morphological picture of the large intestine lesion in ozone-induced colitis correlates with the changes typical of inflammatory bowel disease in humans. The positive effect of ozone in ozone-induced colitis was driven by its anti-inflammatory properties through the activation of Nrf2 and by its antioxidant properties through the inhibition of Keap1

Time course of lymphocyte profile after femoral bone fracture

Isolated fractures of femur account for more than 10% of all road traffic injuries. Traumatic injury of femoral bone triggers a cascade of interrelated neuroendocrine reactions at systemic level, primarily at the hypothalamic-pituitary-adrenal axis, systemic response of immune system, initiated by release of tissue degradation products, cytokines and other mediators of damage into systemic blood circulation. Specific cellular reactions in response to traumatic injury to bone tissue include both innate and adaptive immune responses. In this regard, there is still scarce information on changes in blood lymphocyte subpopulations observed after closed isolated fracture of the femoral diaphysis at the middle third, before and after surgery. The aim of the present study was to evaluate the subpopulations of peripheral blood lymphocytes following closed isolated fracture of the femoral diaphysis with bone displacement in thecourse dynamics of surgical treatment, thus being required for studies in pathogenesis, development of diagnostic criteria and creating innovative treatment approaches. The study included 20 apparently healthy men and 36 men with closed isolated fracture of the femoral diaphysis of the middle third (32A and 32B, by AO/ASIF clinical classification, coded according to ICD-10 S72.3). The exclusion criteria were as follows: exacerbation of chronic comorbidities, diseases of lymphatic system and haematopoietic organs, oncological diseases, and evidence of osteoporosis. The spectrum of blood lymphocyte subsets was assessed on days 5, 7 (immediately after surgery) and on day 18 after closed isolated fracture of femoral diaphysis. We have found that, on the day 5 after IPBC along with leukocytosis in peripheral blood, the number of T-regulatory cells, cells with markers of early (CD25+) and late activation (HLA-DR+) proved to be increased, whereas representation of NK cells was decreased. On the day 7 after IPBC and immediately after surgery, leukocytosis persisted in blood, along with increased number of T-regulatory cells, CD3+ cells with early and late activation markers. On the day 18 after closed isolated fracture of the femoral diaphysis, the total numbers of leukocytes, T-lymphocytes, T-helpers, T-regulatory cells, T cells with an early activation marker are restored in peripheral blood, whereas the number of T-lymphocytes expressing HLA-DR+ molecules showed a significant increase

Immunotropic effects of vitamin D3 in original rectal suppositories in experimental ulcerative colitis

Increased incidence of ulcerative colitis (UC) is a prerequisite for searching new therapeutic approaches, primarily with an opportunity of site-directed impact on the colon lesion. UC pathogenesis is associated with dysregulated immune response, and limited effectiveness of basic therapy for the disorder. Vitamin D3 exhibits antioxidant, anti-inflammatory, immunomodulatory and other properties, it has been shown to be effective in some autoimmune diseases, thus prompting us to study its effect on immune status in UC. We aimed for studying the effect of vitamin D3, as a component of original rectal suppositories, upon clinical course and indexes of immune status in experimental UC. UC in rats was modeled with 3% oxazolone solution. The vitamin D3-containing suppositories (1500 IU) weighing 300 mg were administered per rectum every 12 hours for 6 days. On days 2, 4 and 6 of UC, the clinical features were assessed as well as blood leukocyte counts, numbers of CD3+, CD45RA+; absorbing and NBT-reducing abilities of blood neutrophils were determined; IgM, IgG, IL-6 and IL-8 concentrations in serum were also studied.The DAI index increased in non-treated UC, along with raised neutrophil numbers in blood, their absorption and NBT-reducing activity was also increased, the total number of lymphocytes, including CD3+, CD45RA+ became higher, serum concentrations of IgM, IgG, IL-6, IL-8 increased. Local use of vitamin D3 in UC reduces DAI parameters, causes decrease in blood neutrophil counts, reducing and partially restoring absorptive and NBT-reducing abilities of neutrophils, decline of total lymphocyte counts in blood, partially restoring the CD3+ and CD45RA+ numbers, causing decline and partial restoration of serum IgM, IgG, IL-6, IL-8 concentrations. An association between clinical signs and indexes of immune status in UC was established under the conditions of vitamin D3 use. Conclusions: The protective effect of vitamin D3 in UC can be mediated by its antioxidant effect, changes in production of immunoregulatory cytokines, modulation of Th1-, Th2-, Th17-dependent reactions and Treg activity, being a pre-requisite for further studies to clarify the mechanism of vitamin D3 immunotropic action in UC,with an opportunity of using it in clinical practice

Current therapeutic approaches to haemostasis correction in covid-19: a systematic review

Background. The coronavirus disease 2019 (COVID-19) caused by the SARS-CoV-2 virus has swept across countries worldwide. Despite an unprecedented volume of research, few drug therapies have been proved effective. The lack of evidence-based strategies entailed many practical treatments. Hypercoagulability observed in COVID-19 patients has sparked a debate in the medical community on therapeutic value of anticoagulants.Objectives. A review of up-to-date evidence supporting the therapeutic effect of unfractionated and low molecular-weight heparin as anticoagulant in treatment for COVID-19. Methods. Russian-language and foreign literature was mined in the RSCI, Scopus, PubMed, medRxiv and eLibrary databases for the years 2020–2021, with considering selected impactive publications within 1991–2019 as well. The query keywords were COVID-19, heparin [гепарин], hemostasis [гемостаз], thromboembolism [тромбоэмболия]. Peer-reviewed scientific journals received priority. Content and descriptive analytics were used as research tools.Results. The review surveyed 84 literature sources, with 51 articles selected for downstream analysis. We highlight usage of heparin and its fractions in treatment for COVID-19 and preclinical evidence verifying the antiviral and anti-inflammatory properties of heparin and synthetic heparin-like drugs in COVID-19. The known and plausible side effects demanding additional prospective randomised controlled trials on anticoagulant application in COVID-19 are reviewed, with an assessment of oral direct-acting anticoagulant drug efficiency.Conclusion. Drug-based therapies for haemostasis correction in COVID-19 are currently limited. The paucity of evidence warrants heparin usage as a safer therapy in acute COVID-19 compared to oral anticoagulants. However, the balance of its potential benefits vs. risks must be observed. The benefits and risk uncertainty in heparin treatment require randomised clinical trials and further studies to evaluate safety of direct-acting oral anticoagulants after the patient’s discharge in COVID-19

Melatonin in the dermal film limits the blood lymphocyte death in experimental thermal trauma

According to WHO data, about 11 million people need medical care after burns every year. In the overall structure of burns, the share of thermal trauma (TT) is 80%. Lymphocytopenia in TT is a risk factor for infectious complications and limited repair, and the development of new tools for TT therapy using dermal films is demanded in combustiology. The aim of the study was to evaluate changes in blood lymphocyte parameters, i.e., quantitative composition and their death during experimental thermal damage under the influence of the originally developed dermal film with melatonin (MT) in 49 inbred rats. The grade IIIA TT of 3.5% body surface was modeled by contact with boiling water for 12 s. Dermal films based on sodium carboxymethylcellulose supplemented with MT at a concentration of 0.005 g/g were applied daily for 5 days. The total numbers of lymphocytes, CD45RA+ and CD3+ cells, counts of lymphocytes with signs of partial necrosis, early and late apoptosis were assessed in blood. Relative decrease in the area and rate of the burn wound epithelization were also calculated. In animals with TT, the number of blood lymphocytes decreased on days 5, 10 and 20, including CD45RA+ and CD3+, along with increased amounts of lymphocytes with signs of necrosis, late and early apoptosis. By the term of 20 days, the burn wound area was reduced by 11.5%. Usage of dermal films with MT increased the amount of CD3+ cells in blood on days 5 and 10, CD45RA+ on days 5, 10 and 20, being associated with decreased number of lymphocytes showing signs of early apoptosis on days 5, 10 and 20, as well as features of necrosis and late apoptosis on days 5 following TT, accelerates the healing of a burn wound on days 5, 10 and 20 after TT. with a 20 cent reduction of its area by the day 20. Epithelization rate of the burn wound when applying MT-supplemented dermal film on days 5, 10 and 20 increases, along with higher amounts of CD3+ in the blood, and reduced counts of lymphocytes with signs of early apoptosis

Immunological aspects of the use of melatonin in experimental thermal trauma

The prevalence of thermal trauma, the high risk of infectious and non-infectious short- and long- term complications, and the limited effectiveness of the therapeutic approaches used are prerequisites for the search and pathogenetic justification of new therapies, among which the endogenous homeostasis regulator with pleiotropic properties melatonin attracts attention.The aim of the work is to investigate the immunological aspects of intraperitoneal use of melatonin (MT) in experimental thermal trauma (TT).The work was performed on 158 rats of the Wistar line, grade III TT and a relative area of 3.5% were simulated by skin immersion in water at 98-99 °C for 12 s. MT was administered intraperitoneally daily at a dose of 10 mg/kg for 5 days. The quantitative composition of blood cells was evaluated on a hematological analyzer. Plasma concentrations of IL-4, TNFa, IFNg, and CRP were determined on an automatic enzyme immunoassay using rat-specific test systems, and MT by capillary electrophoresis.With experimental TT, against the background of a progressive increase in the number of leukocytes in the blood from 5 to 20 days due to neutrophils, monocytes, basophils, the number of lymphocytes decreases. With TT, the concentration of CRP increases in serum on days 5 and 10. The content of TNFa, IL-4 increases on days 5, 10 and 20 in the absence of significant changes in the concentration of IFNg. The concentration of serum MT does not change significantly. Intraperitoneal use of MT in TT leads to a partial restoration of the number of lymphocytes in the blood on day 5. Evaluation of the cytokine profile in serum revealed a decrease in the concentration of TNFa on days 10 and 20, no significant changes in the concentration of IL-4 and IFNg were recorded, the concentration of CRP decreased on day 5. The concentration of serum MT increases by 5 days.With TT on the 5th, 10th, 20th day of the experiment, the number of neutrophils, monocytes, basophils in the blood increases, decreases – lymphocytes, the serum content of CRP, TNFa, IL-4 increases, the content of IFNg and melatonin does not change. Intraperitoneal use of MT in TT partially restores the number of lymphocytes in the blood, the concentration of CRP, TNFa. A decrease in serum concentrations of TNFa and CRP in TT under the conditions of MT use suggests a limitation of the acute phase response as a consequence of the antioxidant, anti-inflammatory effect of MT, which can accelerate healing and reduce the area of the lesion of TT

Effect of systemic ozone use on oxidative modification of lipids and proteins in the colon in experimental colitis

The aim of the work was to study the clinical status, the content of lipid peroxidation products, oxidative modification of proteins in the lesion of the colon in oxazole-induced colitis (OIC) under conditions of intraperitoneal application of ozone.Цель работы – изучить клинический статус, содержание продуктов перекисного окисления липидов, окислительной модификации белков в очаге повреждения толстой кишки при оксазолон-индуцированном колите (ОИК) в условиях внутрибрюшинного применения озона

Morphological and immunohistochemical changes in oxazolone-induced ulcerative colitis under ozone therapy

The aim of the study was to investigate the effect of intraperitoneal and rectal ozone therapy on the morphology and morphometry parameters of the damage zone in oxazole-induced ulcerative colitis in the dynamics of the experiment.Цель исследования – изучить влияние внутрибрюшинной и ректальной озонотерапии на морфологию и показатели морфометрии зоны повреждения при оксазолон-индуцированном язвенном колите в динамике эксперимента