Innovative Biosystems and Bioengineering

Cord Blood as a Corrector of Hematological and Cytomorphological Characteristics of Lymphohemopoietic Organs in Experimental Atopic Dermatitis

Hanna Koval — 2023-09-11

Background. Atopic dermatitis (AD) is an inflammatory skin disease caused by a shift in the interaction between cells of the immune system and the skin. Steroid anti-inflammatory drugs used to treat AD often lead to numerous side effects. Therefore, the development of new natural multifunctional preparations with immunomodulatory activity is of great importance.

Objective. To determine the changes in indices of the leukocytes cluster of peripheral blood and determine the peculiarities of cytomorphological changes in the spleen and lymph nodes (LN) of rats with induced AD before and after administration of cryopreserved (cHCBL) and lyophilized (lHCBL) forms of human cord blood leukoconcentrate.

Methods. The experiments were conducted using 6-month-old Wistar rats. AD was induced by topically applying a 5% alcohol-acetone solution of dinitrochlorobenzene (DNCB) to the skin of the rats' back for 21 days. Each form of the preparation (cHCBL and lHCBL) was administered intraperitoneally in a 0.5 ml dose containing 5´10⁶ cells, one day after the final DNCB treatment. The following parameters were assessed in the blood: leukocyte count and the neutrophil-to-lymphocyte ratio (N/L index). Сytomorphological characteristics were determined in the spleen and inguinal LN.

Results. In AD, specific changes in the cytomorphological characteristics of the lymphohematopoietic complex (LHС) structures, namely the spleen and LN, were observed alongside alterations in the leukocyte cluster indices. The most notable changes in the spleen included the dominance of the white pulp over the red pulp and the prevalence of follicles of stages II and III of development in the LN. Animals with AD displayed pronounced leukocytosis and an increased N/L index in their blood. The study demonstrates the positive corrective effects of cHCBL and lHCBL on the morphological characteristics of LHC and the specified blood parameters.

Conclusions. The effectiveness of the cHCBL and lHCBL use for correcting indices of leukocytes cluster of peripheral blood and the structural organization of lymphohemopoietic organs in the development of AD have been proven.

Potential Alphavirus Inhibitors From Phytocompounds – Molecular Docking and Dynamics Based Approach

Maneesha Sharma — 2023-09-20

Background. Alphaviral diseases are an economic burden all over the world due to their chronicity and distribution worldwide. The glycoproteins E1 and E2 are important for binding to the surface of the host cell by interacting with the receptors and non-structural proteins named nsP2 and nsP4 are important for the replication of virus, so can be an important drug discovery target.

Objective. We are aimed to explore the in silico interaction between plant-based compounds (phytocompounds) and specific protein targets, such as nonstructural protein nsP4 and glycoprotein E2 of Sindbis virus (SINV), nsP2 and E2 of Chikungunya virus (CHIKV), and glycoproteins E1 and E2 of Ross River virus (RRV).

Methods. A library of phytochemicals from Indian medicinal plants was prepared using databases and converted to 3D structures. Protein structures (nsP2, nsp4, E1, E2) were obtained and refined, followed by molecular docking with AutoDock Vina. Promising ligands were evaluated for properties, cytotoxicity, and mutagenicity, considering drug-likeness and potential issues. Molecular Dynamics simulations assessed complex stability.

Results. We analyzed 375 phytocompounds against these targets using molecular docking, modeling, and molecular dynamics for SINV, CHIKV, and Ross River (RRV) virus proteins. Granatin A has been found to successfully bind to the target sites of SINV nsP4, CHIKV E2, and CHIKV nsP2 with binding affinity values of -16.2, -20.6, and -18.6 Kcal/mol respectively. Further, stability of CHIKV E2 – Granatin A complex was done by performing molecular dynamic simulation and the complex was stable at 60ps.

Conclusions. This research provides valuable insights into the development of effective antiviral drugs against alphaviruses, emphasizing the importance of natural compounds and their interactions with viral proteins. This study might pave the way for further exploration of these small molecules as effective anti-alphaviral therapeutic agents.

Disturbed Cytokine Profile in Adjuvant Arthritis – A Target of the Therapeutic Potential of Dendritic Cells Derived From Cryopreserved Precursors

Hanna Kisielova — 2023-09-21

Background. One of the primary causes of rheumatoid arthritis development is the disruption of the immune system's natural tolerance to its own antigens, leading to an imbalance in the body's cytokine profile. A promising method of correcting such a condition is restoring antigen-specific tolerance, in the formation of which tolerogenic dendritic cells (tolDCs) take part.

Objective. Experimental substantiation of the possibility of correcting the cytokine profile of animals with adjuvant arthritis (AA) by using tolDCs from cryopreserved bone marrow precursors.

Methods. The study was carried out on the CBA/H mice. The development of AA was assessed using a clinical indicator – the arthritis index. The levels of pro- (TNF-a, IL-6, IFN-g) and anti-inflammatory (IL-10, IL-4) cytokines in the blood serum of AA-afflicted animals were measured before and after administration of tolDCs. These tolDCs were obtained from native (NatDCs) or cryopreserved (CryoDCs) using different methods bone marrow mononuclear cells (MNCs). On the 14th day after inducing AA, the animals received intravenous injections of tolDCs (5´10⁵/mouse). One week later, the cytokine levels in the animals' blood serum and the arthritis index were assessed.

Results. Throughout the development of AA, a unidirectional increase in TNF-α and IL-6 levels and a reduction in the content of anti-inflammatory cytokines were observed, which was accompanied by joint swelling in the animals. CryoDCs exhibited a more pronounced corrective effect on both pro- and anti-inflammatory cytokines compared to NatDCs, as evidenced by a decrease in the arthritis index, a clinical manifestation of the pathology.

Conclusions. The possibility of correcting the disturbed cytokine profile and the clinical state of animals during the development of AA through the use of tolDCs derived from cryopreserved MNCs has been proven. Specific cryopreservation conditions for MNCs have been developed, which facilitate the generation of tolDCs from them with a greater capacity, compared to derivatives of native MNCs, to correct the cytokine profile and clinical status of animals with AA.