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Acellular dermal matrix for chronic wound treatment: dry or wet?
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1Department of Histology and Embryology, Ivan Horbachevsky Ternopil National Medical University, Ternopil, Ukraine;
2Department of Medical Biochemistry, Ivan Horbachevsky Ternopil
National Medical University, Ternopil, Ukraine;
3Department of Pathophysiology, Ivan Horbachevsky Ternopil
National Medical University, Ternopil, Ukraine;
4Central Research Laboratory, Ivan Horbachevsky Ternopil
National Medical University, Ternopil, Ukraine;
5Department of Pharmacology and Clinical Pharmacology,
Ivan Horbachevsky Ternopil National Medical University, Ternopil, Ukraine;
*e-mail: shevchukoo@tdmu.edu.ua
Received: 27 May 2026; Revised: 19 June 2026;
Accepted: 27 July 2026; Available on-line: 04 August 2026
Background. Chronic wounds pose a serious clinical and economic burden, and their effective treatment requires the development of new biomaterials that promote tissue regeneration. Acellular dermal matrices (ADMs) represent a promising regenerative strategy; however, their biological properties largely depend on the manufacturing protocol. Objective. To compare the biocompatibility and cytotoxicity of two porcine acellular dermal matrix variants, dry (d-ADM) and wet (w-ADM), and to evaluate the therapeutic efficacy of the biocompatible matrix in an experimental porcine model of chronic full-thickness wounds. Methods. Biocompatibility was assessed in vitro using human umbilical cord-derived mesenchymal stem cells (hU-MSCs) by evaluating cell morphology, viability, proliferation, and lactate dehydrogenase (LDH) release. The therapeutic efficacy of the selected ADM was investigated in a porcine chronic full-thickness excisional wound model and compared with non-adhesive dressings and Promogran Prisma. Wound healing was evaluated after 28 days using histological, immunohistochemical (TNF-α, TGF-β, Ki-67), and morphometric analyses. Results. The wet ADM preserved hU-MSC morphology, viability, and proliferative capacity, whereas the dry ADM induced marked cytotoxicity, increased LDH release, and extensive cell death. In vivo, treatment with w-ADM significantly accelerated wound healing compared with both control dressings, resulting in complete and stable re-epithelialization, minimal inflammatory infiltration, organized collagen remodeling, and improved tissue architecture. Immunohistochemical analysis demonstrated significantly lower expression of TNF-α, TGF-β, and Ki-67, indicating resolution of inflammation and transition toward tissue remodeling and regenerative stabilization. Conclusions. Wet acellular dermal matrix demonstrated excellent biocompatibility and superior regenerative performance, whereas the dry matrix exhibited unacceptable cytotoxicity. Wet ADM represents a promising scaffold for regenerative treatment of chronic wounds and warrants further investigation as a clinically applicable biomaterial, including in combination with mesenchymal stem cell-based therapies.
Effects of pyrrolidinedione-thiazolidinone hybrid molecules on the genotoxicity in vitro
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1Department of Regulation of Cell Proliferation and Apoptosis,
Institute of Cell Biology, National Academy of Sciences of Ukraine, Lviv, Ukraine;
2Molecular Design Center, Danylo Halytsky Lviv National Medical University, Lviv, Ukraine;
3Department of Pharmaceutical, Organic and Bioorganic Chemistry,
Danylo Halytsky Lviv National Medical University, Lviv, Ukraine;
*e-mail: nataliyafiniuk@gmail.com
Received: 25 May 2026; Revised: 17 July 2026;
Accepted: 27 July 2026; Available on-line: 04 August 2026
Background. Pyrrolidinedione-thiazolidinone hybrid molecules represent a promising class of anticancer candidates; however, a comprehensive evaluation of their genotoxic and mutagenic safety profile is essential before further preclinical development. Objectives. This work aimed to investigate the genotoxic potential of pyrrolidinedione-thiazolidinone hybrid molecules Les-6287 and Les-6294 that possessed antineoplastic activity. Methods. The mutagenic potential was evaluated using the Ames bacterial reverse mutation test with Salmonella typhimurium strains TA98 and TA100 in the presence and absence of metabolic activation (S9 fraction). The effects at the chromosomal level were assessed using the Allium cepa anaphase-telophase chromosome aberration assay. Primary DNA strand break induction was quantified using the alkaline comet assay. Results. Les-6287 and Les-6294 at 10 and 100 µM concentrations do not produce a mutagenic activity exceeding 1.6 in either S. typhimurium strain TA98 or TA100, with or without S9-mediated metabolic activation. In the A. cepa anaphase-telophase assay treatment with Les-6287 and Les-6294 did not result in a statistically significant elevation in chromosomal aberration frequency relative to the negative control (2.8%), with observed values ranging from 3.3% to 4.3% across all tested concentrations. No significant alterations in the mitotic index were recorded. The alkaline comet assay revealed no significant increase in primary DNA damage, with percent tail DNA of 1.3-1.9% in treated peripheral blood mononuclear cells. Conclusion. Pyrrolidinedione-thiazolidinone hybrid molecules Les-6287 and Les-6294 at concentrations up to 100 µM do not pose a genotoxic or mutagenic risk under the tested experimental conditions. Further targeted safety assessments are needed prior to progressing Les-6287 and Les-6294 to preclinical evaluation as candidate antitumor agents.
Hemostatic system imbalance in the pre-eclampsia model in rats
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1SI “Institute of Pharmacology and Toxicology, National Academy
of Medical Sciences of Ukraine”, Kyiv;
2Palladin Institute of Biochemistry, National Academy
of Sciences of Ukraine, Kyiv;
3Shupyk National Medical Academy of Postgraduate Education, Kyiv, Ukraine.
*e-mail: anna_shayakhmetova@yahoo.com
Received: 03 June 2026; Revised: 17 June 2026;
Accepted: 27 July 2026; Available on-line: 04 August 2026
Background. Preeclampsia (PE) is a multisystem disorder in pregnancy, that results from placental ischemia, oxidative stress-induced placental damage and placental factors dissemination that cause systemic endothelial dysfunction. Patients with preeclampsia often develop a severe prothrombotic state, the mechanisms of which are not fully elucidated. Objectives. The aim of this study was to assess the informativeness of hemostatic system parameters that characterize the degree of coagulation pathway activation and anticoagulant pathway capacity capacity in experimental preeclampsia model in rats for predicting the development of thrombotic complications in PE. Methods. Wistar females rats were used in the study. To induce a preeclampsia-like syndrome, N(ω)-nitro-L-arginine methyl ester (L-NAME), at a concentration of 0.3 g/l in the drinking water was administered ad libitum from the 8th to the 14th day of gestation. On gestational day 21 the rats were anesthetized and blood samples were collected. Urinary and serum concentrations of creatinine, urea, and uric acid were determined using a fully automated Chemistry Analyzer and commercially available kits. Fibrinogen, soluble fibrin monomeric complexes and protein C concentrations, prothrombin time and antithrombin III activity in the blood plasma were estimated with a standard spectrophotometric methods. Results. Increased plasma fibrinogen, significant shortening of plasma clotting time, decrease in both protein C and AT III and increase in SFMCs plasma levels were detected in rats with experimentally L-NAME induced preeclampsia, indicating not only activation of the coagulation system but also disruption of the balance between procoagulant and anticoagulant pathways. Conclusions. L-NAME-induced PE in rats resulted in a pronounced imbalance of the hemostatic system and reproduced key pathophysiological features of PE, including excessive procoagulant shift, intravascular thrombin generation and impairment of anticoagulant mechanisms, making this model a valuable tool for studying mechanisms of disease progression. Analysis of hemostasis parameters in this model, indicates that soluble fibrin, protein C and antithrombin III are the most informative markers for assessing the risk of intravascular coagulation during preeclampsia.
Markers of prothrombotic state of patients recovered after COVID-19
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1Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
2Ivan Horbachevsky Ternopil Medical University, Ternopil, Ukraine;
3International Research and Innovation in Medicine Program, CSMC, Los-Angeles, CA, USA;
*e-mail: d.korolova@gmail.com
Received: 08 May 2026; Revised: 18 July 2026;
Accepted: 27 July 2026; Available on-line: 04 August 2026
Background. In recent period the attention of scientists and doctors was focused mainly on methods of COVID-19 treatment. However, clinical course of coronavirus disease strongly affects blood coagulation with the tendency for venous and arterial thrombosis, leading to long-term imbalance between coagulation and anticoagulation systems, which can cause serious danger for health of post-COVID-19 (PCD) patients. Objective. In our work we focused on the analysis of the molecular markers of hemostasis system activation in blood plasma of patients recovered after COVID-19. Methods. Blood plasma samples of 119 seropositive patients recovered from COVID-19 were analysed. The patients were divided into a high-risk group (HRG, n = 163) and a non-risk group (NRG, n = 28) according to WHO recommendations. Patients were examined in a recovery time scale of 1-30; 31-60 days and more than 2 months. Blood plasma samples of seronegative healthy volunteers were collected as the reference group (n = 19). The concentration of soluble fibrin (SF) and D-dimer was measured with sandwich ELISA, generation of activated protein C (PC) and prethrombin-1 (Pre-1)) level with assay using specific chromogenic substrates, and fibrinogen concentration with spectrophotometric method after initiation of coagulation by Ancistron-H. Results. Concentration of fibrinogen, SF and Pre-1 were increased in a blood plasma of patients of NRG and HRG groups in comparison to the reference group. Increasing of fibrinogen and Pre-1 concentrations correlated with COVID-19 stage. The fibrinogen and SF concentrations remained high more than 2 months after recovery in HRG group and elevated after 2 months after recovery in NRG group. D-dimer and protein C levels were elevated in 8% and 11% of PCD patients respectively. Conclusions. Long-term prethrombotic state was more expressed in HRG group of PCD patients. Monitoring of the main coagulation markers can indicate the risk of intravascular thrombus formation in PCD patients. SF, Pre-1 and elevated fibrinogen concentrations were selected as useful thrombosis markers in PCD patients.
Estimation of circulating microRNAs as biomarkers in early stages of chronic kidney disease
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1College of Veterinary Medicine, Al-Qasim Green University,51013, Babylon, Iraq;
2College of Science, Al-Qasim Green University, 51013, Babylon, Iraq;
*e-mail: hamza14shukri72@gmail.com
Received: 29 April 2026; Revised: 24 June 2026;
Accepted: 27 July 2026; Available on-line: 04 August 2026
Background. Chronic kidney disease (CKD) is a progressive disorder that affects 10–13% of the worldwide population. The conventional biomarkers, like serum creatinine and estimated glomerular filtration rate (eGFR), have poor sensitivity in detecting early stage CKD, especially as it relates to Type 2 Diabetes Mellitus (T2DM). Recent studies suggest that circulating microRNAs (miRNAs) are sensitive molecular markers of renal pathology. Objectives. This study quantified the serum levels of miR-21, miR-155 and miR-192 in early-stage CKD patients with or without co-existing T2DM and determined the diagnostic performance of these microRNAs compared to cystatin C and eGFR. Methods. A case-control study was conducted with a total of 100 participants classified into three groups; healthy controls (n = 30), CKD patients with T2DM (n = 35) and CKD patients without T2DM (n = 35). The expression of serum miRNA was measured with real-time quantitative PCR using the 2⁻ΔΔCt method, U6 snRNA as internal reference. Cystatin C was determined by particle-enhanced immunonephelometry; eGFR was calculated using the CKD-EPI equation. Statistical analyses were performed using Kruskal-Wallis test, Spearman partial correlations, receiver operating characteristic (ROC) analysis, and multivariate logistic regression. Results. Significant upregulation of all three miRNAs in both CKD groups (P < 0.001) with a differing magnitude of fold-change compared with controls and the highest amplification for miR-192 was detected. ROC analysis identified miR-192 to be the best performing single biomarker (AUC = 0.937; 95% CI: 0.880–0.978). Spearman partial correlation networks showed strong positive relations of miRNAs with cystatin C (ρ = 0.74–0.80) and stronger negative associations with eGFR (ρ = −0.69 to −0.78). Conclusions. Circulating miR-21, miR-155 and miR-192 are promising minimally invasive biomarkers better than conventional markers for early CKD detection. Cystatin C was co-measured with it, yielding a highly accurate diagnostic panel especially in the diabetic nephropathy sub-phenotype. Clinical applicability requires validation in large multicentre cohorts.
Cation and substrate specificity of the sodium pump and the mechanokinetics of the “contraction–relaxation” process in smooth muscle under the action of calix[4]arene C-1130
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1Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv, Ukraine;
*e-mail: o.maliuk@biochem.kiev.ua;
2Educational and Scientific Institute of High Technologies,
Taras Shevchenko National University of Kyiv, Kyiv, Ukraine;
3Institute of Organic Chemistry, National Academy of Sciences of Ukraine, Kyiv, Ukraine
Received: 29 April 2026; Revised: 24 June 2026;
Accepted: 27 July 2026; Available on-line: 04 August 2026
Background. Plasma membrane Na+,K+-ATPase plays an important role in maintaining ionic homeostasis and regulating myometrial contractility. Its inhibition may alter uterine smooth muscle responses to uterotonic stimulation. Objective. This study aimed to determine the mechanism by which calix[4]arene C-1130 inhibits Na+,K+-ATPase activity and to evaluate its effects on oxytocin-induced myometrial contractions. Methods. Na+,K+-ATPase activity was studied in suspensions of perforated plasma membranes isolated from myometrial cells. The kinetic parameters of ATP hydrolysis were determined at different ATP and Mg2+ concentrations. The effects of C-1130 on oxytocin-induced isometric and isotonic contractions were examined in myometrial preparations with preserved endometrium. Results. Calix[4]arene C-1130 decreased the maximum rate of Na+,K+-ATPase-mediated ATP hydrolysis without significantly affecting the apparent affinity or cooperativity parameters for ATP and Mg2+. At a concentration of 10 μM, C-1130 enhanced oxytocin-induced isometric myometrial contraction and increased the normalized maximal rates of both contraction and relaxation phases. Under isotonic conditions, C-1130 increased the velocity and enhanced the power characteristics of muscle contraction. Conclusions. Calix[4]arene C-1130 inhibits myometrial Na+,K+-ATPase through a noncompetitive mechanism. The enhancement of oxytocin-induced contractile muscle activity is likely associated with disruption of ionic homeostasis in myocytes caused by inhibition of the plasma membrane Na+,K+-ATPase.
Role of fibrinogen and fibrin in initiation of thrombin generation
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Palladin Institute of biochemistry, National Academy of Sciences of Ukraine, Kyiv, Ukraine;
*e-mail: nudovenko26@gmail.com
Received: 07 May 2026; Revised: 27 June 2026;
Accepted: 27 July 2026; Available on-line: 04 August 2026
Background. Thrombin is the key enzyme of blood coagulation that converts fibrinogen into fibrin and activates multiple coagulation factors. However, the temporal relationship between thrombin-mediated fibrin formation and the availability of thrombin for other physiological substrates during the initiation of coagulation remains insufficiently understood. Objective. To investigate the role of fibrinogen and fibrin in regulating thrombin activity during the initiation phase of thrombin generation in plasma. Methods. Thrombin generation was initiated in human plasma using activated partial thromboplastin time (APTT) reagent or Thromborel. Fibrin formation was monitored turbidimetrically at 405 nm. Thrombin amidolytic activity toward the chromogenic substrate S2238 was measured spectrophotometrically at 405 nm. Fibrin polymerization was inhibited with the synthetic peptide GPRP. The effects of plasma dilution and inhibition of fibrin polymerization on the temporal profiles of fibrin formation and thrombin activity were analyzed. Results. During initiation of thrombin generation with both APTT-reagent and Thromborel, the beginning of cleavage of S2238 by thrombin coincided with the end of fibrin clot formation, marked by turbidimetric curve. At 1:3 plasma dilution, fibrin formation lag time was 210 s, amidase activity lag phase was 410 s. At 1:10 dilution, the lag time for fibrin formation was 73 s, amidase activity was 200 s, indicating that dilution (and lower fibrinogen) accelerates thrombin generation. With 1 mM GPRP, fibrin and amidase activity lag times increased by an order of magnitude. Conclusions. Thrombin generated during initiation of thrombin generation acts primarily on fibrinogen, remaining bound to the molecule throughout its conversion into fibrin and clot formation. Its amidase activity emerges only after protofibrils undergo lateral association into fibrin clot. Once the binding sites in the fibrin clot are saturated, thrombin becomes available for other substrates (FV, FVIII, FXI) and physiological functions.
Purification and physicochemical properties of Bacillus licheniformis 249 protease with elastolytic and fibrin(ogen)olitic activity
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1Danylo Zabolotny Institute of Microbiology and Virology,
National Academy of Sciences of Ukraine, Kyiv;
2Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
3Odesa I. I. Mechnikov National University, Odesa, Ukraine;
*e-mail: alena.gudzenko81@gmail.com
Received: 22 April 2026; Revised: 27 May 2026;
Accepted: 27 July 2026; Available on-line: 04 August 2026
Background. Proteolytic enzymes with a broad substrate specificity are of considerable interest for potential application in biotechnology and medicine. Proteases with specificity for elastin, fibrinogen and fibrinogen are of particular interest, in particular as antithrombotic agents. Marine microorganisms due to adaptive mechanisms to environment could be a potential producers of enzymes capable of maintaining catalytic activity in a wide range of physicochemical conditions. Objective. The aim of this work was to purify the protease produced by Bacillus licheniformis 249, isolated from the bottom sediments of the Black Sea and to study the physicochemical and catalytic properties as well as substrate specificity of enzyme. Methods. The enzyme was obtained from the supernatant of the bacterial culture liquid by precipitation with ammonium sulfate (90% saturation) with subsequent gel-permeation chromatography on Toyopearl HW-65F and ion-exchange chromatography on Toyopearl DEAE-650M. Results. The purified enzyme had a molecular weight of about 30.0 kDa, maximal activity at 37°C, pH 8,0 and 11,0 and was characterized by high elastolytic, fibrinogenolytic and fibrinolytic activities with a values of 2697.3; 1620 and 1615 U/mg protein, respectively. Electrophoretic analysis of fibrinogen hydrolysis products revealed selective cleavage of the Aα chain, which allows us to attribute the studied enzyme to α-fibrinogenases. Conclusion. The combination of elastolytic and fibrin(ogen)olytic activities, as well as stability in a wide range of physicochemical conditions, indicate the promising potential of this enzyme for use in medicine and various biotechnological processes.