V.I.Shumakov Federal Research Center of Transplantology and Artificial Organs

Objective: The formation of a tissue equivalent based on injectable form of microdispersed scaffold— microparticles of decellularized porcine cartilage (DecCp)—seems to be a promising technology for repairing cartilage tissue defects. The aim of this work was to obtain and comparatively study a tissue-engineering construct (TEC) based on DecCp microparticles and mesenchymal stromal cells (MSCs) under static conditions and in perfusion bioreactor. Materials and methods: The decellularization process included freeze-thaw cycles (–196°C/+37°C), the use of surfactants (Triton X-100 and sodium dodecyl sulfate), as well as DNase treatment. The morphology of the surface and the nearest subsurface layer of the samples was studied using scanning electron microscopy. Each TEC consisted of 5 × 105 MSCs and 5 mg DecCp. Results: It was found that, compared with static conditions, the cultivation of MSCs on DecCp microparticles in a perfusion bioreactor for 14 days allows increasing the proliferative activity of cells with subsequent chondrogenic differentiation, as evidenced by the ability of the cellular component of cartilage to synthesize extracellular matrix (ECM), characteristic of cartilage tissue, histochemical analysis of which revealed the presence of collagen and glycosaminoglycans (GAG). Conclusion: The possibility of forming cartilage TECs based on DecCp and MSCs under 3D cultivation conditions both under static conditions and in a perfusion bioreactor was shown. Cultivation of MSCs on DecCp under flow conditions at a rate of 0.5 mL/min contributed to an increase in cell proliferative activity compared with static conditions, and also supported the ability of cells to synthesize ECM, characteristic of cartilage tissue, histochemical analysis of which revealed the presence of total collagen and GAG, which may be evidence of chondrogenic differentiation of MSCs.

Background : The annually increasing number of surgical procedures worldwide makes the issue of adequate perioperative fluid therapy controversial. In the operating room, patients are exposed to various anesthetic and surgical factors, while their hemodynamics constantly change due to surgical trauma, blood loss, and other factors. Therefore, the question of the necessity of intraoperative fluid therapy remains particularly critical. Objective : To analyze various methods for assessing a patient’s volume status and predicting their response to fluid therapy. Materials and methods : The publication search was conducted independently by two researchers using databases and electronic libraries, including PubMed (MEDLINE), Cochrane Library, Google Scholar, and eLibrary. Following the selection process, the 40 most informative publications were identified, which formed the basis of this review. Results : This review describes static hemodynamic parameters, functional tests (mini-fluid challenge, passive leg raise, and end-expiratory occlusion test), and dynamic diagnostic methods (pulse pressure variation, systolic pressure variation and stroke volume variation, inferior vena cava collapsibility index, left ventricular outflow tract velocity time integral, respiratory variation of peak systolic velocity, corrected carotid flow time, and internal jugular vein distensibility index). Numerous studies have demonstrated the superior predictive value of dynamic methods compared to static parameters; however, no universal diagnostic method currently exists. Conclusion : The search for the optimal combination of methods to assess a patient’s volume status and predict their response to fluid therapy remains a subject for further research.

The aim of this study was to conduct a comparative analysis of the bioregulatory role of mesenchymal stem cells (MSCs) in the liver under physiological conditions, in acute and chronic injury with fibrotic remodeling, and during therapeutic correction by implanting exogenous MSCs from healthy tissues into the body. The analysis showed that hepatic MSCs maintain structural homeostasis by interacting with tissue myofibroblasts and migrating immune cells. In acute liver injury that does not deplete adaptive reserves, hepatic (resident) MSCs regulate tissue homeostasis.. Chronic injury that depletes adaptive reserves activates both immune cells and hepatic MSCs, leading to liver inflammation and the transdifferentiation of MSCs into myofibroblasts. These activated fibroblasts overproduce extracellular matrix components, thereby driving liver fibrosis progression. Exogenous apoptotic MSCs from healthy auto- or allogeneic tissues, when administered in cases of chronic liver injury, can compensate for deficient regulatory factors and restore metabolic regulation and structural homeostasis through their paracrine and trophic activity. Their therapeutic potential is maximized when their regulatory properties are enhanced prior to administration and when applied in recipients without irreversible liver injury.

Mesenchymal stem cells (MSCs) are known to assist liver regeneration. In this study, we show a dose-dependent mode of recovery from liver fibrosis after intravenous injections of MSCs. Male Wistar rats experienced a 42-day-long modeling of liver fibrosis via CCl4 poisoning and received either a single injection of 2.5 × 106 MSCs on Day 3 after the last CCl4 dose or two MSC injections on Days 3 and 10. We dynamically monitored levels of liver cytolysis markers and cytokines in the venous blood and performed a histological study of Mallory-stained liver sections. All experimental groups experienced a nearly complete recovery of biochemical markers up to 4 weeks after the end of CCl4 administration, although we observed anti-inflammatory changes in the cytokine levels only in animals treated by two MSC injections. Histological study revealed minor signs of liver damage up to Day 90 in animals receiving two MSC doses with worse pathology in those who received a single MSC dose. Morphometric values stayed consistent with visual data, demonstrating a significantly larger number of binuclear hepatocytes, a smaller number of false lobules, and a lesser area of connective tissue proper in animals treated by two MSC injections. Our results reflect MSC grafting in applied doses to affect liver fibrosis in a dose-dependent mode. These findings provide a deeper understanding of MSC action in liver fibrosis, and the doses applied may serve as a milestone for further studies in humans.

Objective: to investigate the biodegradation of natural silk-based tissue scaffolds (NS-TS) under in vitro and in vivo conditions, assessing their potential for tissue engineering applications.Materials and methods. Two types of NS-TS, Fibroplen-Atlas and Fibroplen-Gas, along with their modified versions, were analyzed. In vitro biodegradation was assessed in Fenton’s solution, while in vivo studies were conducted on rats, with histological and morphometric analysis of the implants at 4, 14, and 56 days post-implantation.Results. In vitro biodegrada- tion studies showed that Fibroplen-Gas completely degraded in <15 days, whereas Fibroplen-Atlas persisted for up to 45 days. In vivo analysis showed gradual resorption of all scaffolds, with Fibroplen-Gas exhibiting more pronounced degradation. Histological examination revealed a macrophage response, formation of foreign-body giant cells, and signs of implant vascularization. Morphometry confirmed a reduction in filament cross-sectional area, particularly in modified samples.Conclusion. Modifications of NS-TS influence their biodegradation rate, inflammatory response, and vascularization.

Acetylated tubulin is a hallmark of flagellar stability in spermatozoa, and studies have demonstrated the ability of CDYL to function as a tubulin acetyltransferase in spermatozoa. Of note, germline conditional knockout of Cdyl can lead to asthenoteratozoospermia and infertility in male mice. However, the role of CDYL gene in human fertility remains uncharacterized. Data were collected through in silico analysis for an infertile man with asthenoteratozoospermia of Han Chinese descent by performing whole-exome sequencing. Light and electron microscopy were used to characterize the sperm cells of the proband, and the pathogenicity of the genetic factors was determined by functional experiments. To overcome fertility problems, intracytoplasmic sperm injections were performed in the couple. Here, we recruited an infertile proband, born to first-cousin parents, displaying idiopathic asthenoteratozoospermia. Whole-exome sequencing identified a splicing mutation (c.103+1G>A) in CDYL, recessively cosegregating in the family. In vitro minigene assays demonstrated that the mutation resulted in aberrant alternative splicing. We found that CDYL co-localizes with Ac-tubulin along the flagella of human spermatozoa. In addition, the expression of Ac-tubulin was severely reduced in spermatozoa from the patient with CDYL mutation. Disruption in CDYL results in thin mid-piece related abnormal flagella morphology and decreased sperm motility. The primary manifestation of sperm ultrastructural abnormalities under the electron microscope is primarily characterized by disorder of axonemal protein complex and anulus. We demonstrated that a homozygous CDYL splicing mutation specifically induces a decrease in microtubule acetylation, resulting in thin mid-piece related asthenoteratozoospermia, providing a novel marker for genetic counseling and diagnosis of male infertility.

Bile leakage is a common complication after liver resection. It often requires repeated interventions or surgery and prolongs the patient’s recovery. The aim of the study was to assess the effectiveness of the leakage test with fat emulsion (the White Test) in preventing postoperative biliary complications. A multicenter (3 hospitals) randomized controlled trial was performed from February 2011 to May 2016. The trial involved only the patients scheduled for major hepatectomies. After liver transection and control of biliary tree leak-proofness, the patients were randomized into two groups—with and without applying the White Test. A comparative assessment of all the White Test participants was conducted. Forty-three patients formed the study group, and 36 patients were included in the control group. The White Test revealed sites of bile leakage (the positive White Test) in 37.2% (16/43) of the patients in the study group. These leakage sites were sealed intraoperatively. One of those patients (6.2%; 1/16) still developed bile leakage after surgery. Bile leakage was still observed in 7.4% (2/27) of patients after the negative White test. The incidence of postoperatively revealed bile leakage in the study and control groups did not have a statistically significant difference: 7% (3/43) and 8.3% (3/36), respectively. All bile leaks were grade B. This study demonstrated that the White Test did not provide any benefit in preventing postoperative bile leakage; therefore, other methods, such as ICG, should be further investigated.

Patients with acute kidney disease (AKD) may need treatment by hemodialysis and/or apheresis, which is often started in haste due to the life-threatening condition. To illuminate patients' lived experiences of being in acute need of such therapy due to AKD, we performed semi-structured interviews analyzed with qualitative content analysis. Eleven patients out of 104 were eligible and completed interviews. Six main categories were found - shock and emotional chaos, loneliness in an unfamiliar situation, a need for tailored information, emotional support from next of kin, the dialysis and/or apheresis experience varied, and the disease-conditioned life. An overarching theme emerged as, 'Will I survive? Will I be cured?' Clear and repeated information and personalized contact were requested to reduce anxiety and give hope during a time of crisis.