Editorial
Objective: to monitor current trends and developments in organ donation and transplantation in the Russian Federation based on the 2019 data. Materials and methods. Heads of organ transplant centers were surveyed. Data obtained over years from federal subjects of the Russian Federation and from organ transplant centers in the country were analyzed and compared. Results. Based on data retrieved from the 2019 Registry, only 46 kidney, 31 liver and 17 heart transplant centers were functioning in Russia. In 2019, there were 6,878 potential recipients in the kidney transplant waitlist. This represents 13.7% of the 50,000 dialysis patients in the country. Donation activity in 2019 reached 5.0 per million population; multi-organ procurement rate was 71.6%; 2.9 organs on average were procured from one effective donor. In 2019, there were 10.0 kidney transplants per million population, 4.0 liver transplants per million population and 2.3 heart transplants per million people. Same year, the number of transplant surgeries performed in Russia rose 10.7% from the previous year. Moscow and Moscow Oblast alone have 13 functioning organ transplantation centers. They account for half of all kidney transplant surgeries and 70% of all liver and heart transplants performed in the country. Organ recipients in the Russian Federation have exceeded 16,000 in number. Conclusion. Organ transplantations in Russia keep on increasing – 10–15% per year. Donor and transplant programs are also becoming more effective and efficient. However, the demand for organ transplants far exceeds the current supply of available organs in the Russian Federation. Peculiarities of the development of organ donation and organ transplantation in Russia in 2019 were associated with some factors, such as structure and geographical location of transplant centers, waitlisting of patients, funding sources and amount, and management of donor and transplant programs. The national transplantation registry will be developed taking into account new monitoring and analysis challenges.
Clinical Transplantology
Objective: to reflect on a 5-year experience in liver transplant surgery at the Rostov Regional Clinical Hospital. Materials and methods. Liver transplant was performed in Rostov Oblast in July 2015 for the first time. There were 52 liver transplant surgeries performed in the region by the end of February 2020. Cirrhosis due to viral hepatitis is the leading indication for liver transplantation in 33.3% of patients. The average age of recipients was 43.5 ± 15.8 years. Male recipients accounted for 59.6% of cases. Nine recipients got liver transplants from blood relatives, while 43 recipients received an organ from post-mortem donors. For two patients, liver graft was obtained by splitting the liver into two lobes using the in situ split technique. Results. The average duration of surgery was 5.14 ± 1.92 hours. Blood loss during surgery did not exceed 1400 ml. Up to 93% of lost blood was recovered using the reinfusion system. The need for red blood cell transfusion was observed in 48.1% of cases. Fresh frozen plasma was transfused in all cases. Early postoperative complications were observed in 15 patients (29.4%), and some of them had several complications simultaneously. Biliary and vascular complications, which were eliminated by minimally invasive methods and open surgeries, had a significant influence on liver transplant outcome. In-hospital mortality was 5.6%. The causes of death were intra-abdominal bleeding (1), portal vein thrombosis (1) and biliary sepsis (1). Four more people died in the long term after being discharged from hospital: lung cancer (1), graft rejection (1) and fungal sepsis (2). Conclusion. Liver transplant outcome depends on the skills and experience of the specialists implementing this program. Post-transplant in-hospital and long-term mortality depends on the presence and nature of complications, and on the possibility of early treatment.
Objective: to evaluate the clinical efficacy and outcomes of kidney transplants performed using an alternative immunosuppressive therapy protocol that is based on double induction. Materials and methods. We examined 296 cases of kidney transplants performed in 295 patients between January 1, 2004 and December 31, 2018. Based on induction immunosuppressive therapy regimen, the patients were divided into two groups. Group 1 included patients who underwent transplantation from January 1, 2004 to June 30, 2013 and who used the standard induction immunosuppression protocol. Group 2 included patients who did transplant surgeries between the period January 7, 2013 and December 31, 2018 and who received the “double” induction protocol being analyzed. The method of dividing patients into these groups is associated with routine implementation of the analyzed protocol at the transplantation center since July 1, 2013. Results. Graft and recipient survival rates at all follow-up periods were higher in the group of patients who received the “double” induction immunosuppressive protocol than in the standard group. The studied protocol provides initially better and more stable graft function than in standard therapy. This is especially valuable in centers experiencing difficulties in assessing pre-transplant immunological risk. The graft and recipient survival rates achieved by the analyzed protocol are more pronounced in deceaseddonor kidney transplantation. Conclusion. Positive results obtained from retrospective analysis of the protocol under study justify a prospective randomized study.
Objective: to study the indicators of the monocyte-derived component of the immune system in kidney transplant recipients with satisfactory early and delayed renal transplant function. Materials and methods. The study involved 76 kidney transplant recipients. Concentrations of serum creatinine (sCr), serum urea (sUr) and serum cystatin C (sCysC) were measured. CD14+mid/high and CD14+low were isolated from CD14+ monocytes. CD64- and CD86-expressing cell counts were determined for each subpopulation. Immunological examination was performed before surgery, as well as at days 1, 3, 7, 30, 90, 180 and 360 after surgery. Results. There was significant imbalance between the two monocyte subpopulations before transplantation and in the early post-transplant period (first 3 months). By the end of a 6-month follow-up period, the percentage of CD14+ cells had normalized. The dynamics of the subclasses of CD86-expressing monocytes in the post-transplant period is somewhat different from the dynamics of the total count for these monocytes. However, by the end of a 6-month follow-up period, these biomarkers returned to normal for the group of healthy individuals (CD14+mid/highCD86+ p180 = 0.079; CD14+lowCD86+ p180 = 0.789). CD14+lowCD64+ level was significantly higher in the kidney transplant group than in the control group during the entire follow-up period (p0 = 0.0006, p1 = 0.0001, p7 = 0.005, p30 = 0.005, p90 = 0.007, p180 = 0.0002, p360 = 0.001). On the other hand, CD14+mid/highCD64+ count for up to 180 days was not significantly different from that of the control group (p0 = 0.561, p1 = 0.632, p7 = 0.874, p30 = 0.926, p90 = 0.912), with subsequent significant increase by day 360 of follow-up (p180 = 0.01, p360 = 0.003). We observed a negative correlation between CD14+lowCD86+ level at day 0 and sCr levels at day 7 (r = –0.4; p = 0.008) and day 360 (r = –0.34; p = 0.042) and sCysC level at day 7 (r = –0.57; p = 0.014). A negative correlation was also found between CD14+lowCD86+ at day 1 and sCr levels at day 7 (r = –0.4; p = 0.005) and day 360 (r = –0.39; p = 0.02). There was positive correlation between the CD14+lowCD64+ subpopulation index at day 0 and sCr (r = 0.54; p = 0.008) and sCysC (r = 0.6; p = 0.008) levels at day 7, and also between the CD14+lowCD64+ count at day 1 and sCr (r = 0.55; p < 0.0001) and sCysC (r = 0.58; p = 0.004) levels at day 7. CD14+mid/highCD64+ at day 0 negatively correlated with sCysC level at day 360 (r = –0.85; p = 0.015), while CD14+mid/highCD64+ at day 7 positively correlated with sCysC level at day 360 (r = 0.50; p = 0.016). Conclusion. Before transplant surgery, CD14+mid/high, CD14+mid/highCD86+ , and CD14+lowCD86+ counts were reduced, while those of CD14+low, CD14+mid/highCD64+ and CD14+lowCD64+ were increased. By the 6-month follow-up, all these subpopulations except CD14+mid/highCD64+ had reached values for healthy people. Positive correlation between CD14+mid/high, CD14+lowCD64+ , CD14+mid/highCD86+ , CD14+mid/highCD64+ counts in the early post-transplant period and sCr/sCysC levels in long-term follow-up, as well as negative correlation between CD14+low, CD14+lowCD86+ counts in the early post-transplant period and sCr/sCysC levels in long-term follow-up can serve as a predictor of renal graft function.
Objective: to assess the prevalence of hyperparathyroidism (HPT) and the factors affecting its development in kidney transplant recipients. Materials and methods. The single-center observational cohort study included 97 kidney transplant recipients – 40 men, 57 women, age 50 ± 9 years. Inclusion criteria: more than 12 months of post-transplant period, 3 months of stable renal transplant function. Non-inclusion criterion: therapy with vitamin D, with its alternatives or with cinacalcet. Dialysis ranged from 0 to 132 months (median 18); 46% of patients had pre-operative secondary HPT. A comprehensive laboratory study included evaluation of serum concentrations of parathyroid hormone (PTH), 25-OH vitamin D, calcium, phosphorus, magnesium, total alkaline phosphatase (ALP) activity, albumin, creatinine and daily proteinuria. At the dialysis stage, the target PTH range of 130–585 pg/ ml was used, in the post-transplant period – ≤130 pg/ml. Glomerular filtration rate (eGFR) was calculated using the CKD-EPI (Chronic Kidney Disease Epidemiology Collaboration) formula. Results. Patients were divided into two groups based on PTH threshold level (130 pg/ml): the first with HPT (PTH >130 pg/ml, median 203), the second without HPT (PTH ≤130 pg/ml, median 101). Both groups were comparable in terms of gender, age, primary renal disease, dialysis modality, post-transplant follow-up, and immunosuppressive therapy regimen. In group 1 and group 2 recipients, dialysis therapy, pre-transplant median PTH level, incidence of reoperation and incidence of immediate renal graft function were 30 (14; 50) and 14 (6; 28) months (p = 0.004), 681 (538; 858) and 310 (182; 556) pg/ml (p < 0.001), 17% and 2% (p = 0.028), 51% and 80% (p = 0.005), respectively. At the time of the study, 72% of group 1 recipients had eGFR <60 ml/min, versus 36% of group 2 (p >< 0.001). Among HPT biochemical parameters, there were differences for ionized serum calcium (1.32 ± 0.07 versus 1.29 ± 0.04 mmol/l, p = 0.017) and ALP activity (113 ± 61 versus 75 ± 19 u/l, p = 0.021). Serum vitamin D in both groups reduced in equal measures – 14 ± 4 and 15 ± 6 ng/ml. Conclusion. Persistent HPT in the long-term post-transplant period reaches 48.5%. Risk factors for its development included dialysis for more than 18 months, pre-operative secondary HPT, repeated kidney transplantation, delayed graft function, and eGFR <60 ml/min.
Objective: to study the electrokinetic and aggregation properties, as well as the pro-oxidant and antioxidant processes in red blood cells following kidney transplantation in donors and in recipients in the postoperative period. Materials and methods. Blood from 12 recipients and 5 kidney donors over time – before transplantation, as well as at week 1, months 1, 2, 7, 10 and 12 after surgery, as well as from 8 healthy volunteers who formed the control group. We used microelectrophoresis to measure the electrophoretic mobility of red blood cells, characterizing the electrokinetic properties of cells. Aggregation was calculated microscopically by counting unaggregated red blood cells. Malondialdehyde concentration was measured spectrophotometrically at its absorbance maximum at 530 nm by reaction with thiobarbituric acid. Catalase activity was analyzed by reducing hydrogen peroxide in the sample spectrophotometrically at 240 nm wavelength. The obtained values were compared using the Mann–Whitney U test. Results. Decreased electrophoretic mobility of red blood cells within 2 months after transplantation was associated with increased malondialdehyde concentration and erythrocyte aggregation, decreased catalase activity in kidney recipients, followed by restoration of indicators to the control values. Electrophoretic mobility of red blood cells decreased, while malondialdehyde concentrations increased in donors after surgery. However, the increase was less pronounced than in recipients. The changes indicate that the postoperative period causes changes at the cellular level both in donors and in recipients. This is manifested by decreased stability of erythrocyte membrane structure, which is largely determined by lipid peroxidation processes. At the systemic level, a change in the electrophoretic mobility of red blood cells indicates a stress reaction before and after kidney transplantation in recipients within 2 months after surgery, and in donors in 1–2 months in the postoperative period with gradual increase in the body’s resistance. Conclusion. Kidney transplantation is manifested at the cellular and systemic levels. At the cellular level, there is decreased stability of the membrane structure, which is largely determined by lipid peroxidation processes. At the systemic level, a change in the electrophoretic mobility of red blood cells indicates a stress reaction with gradual increase in the body’s resistance. The data obtained demonstrate changes in the functional properties of red blood cells both in kidney transplant recipients and in donors. These changes need to be taken into account when carrying out therapeutic measures.
Objective: to assess the potentials of using indocyanine green fluorescence angiography in evaluating revascularization of tissue-engineered construct that was obtained from the decellularized biological matrix of primate trachea, including using mesenchymal stem cells, after heterotopic tracheal allotransplantation. Material and methods. Tracheas were obtained from two male hamadryas baboons. After decellularization, 4 cm segments of tracheas were implanted under the lateral part of the latissimus dorsi in two healthy primates, one after recellularization with mesenchymal stem cells (animal 1), and the second without recellularization (animal 2). Immunosuppressive therapy was not performed. Blood flow in the transplanted segment of the trachea was evaluated 60 days after transplantation by surgical isolation of the flap of the latissimus dorsi with the transplanted segment of the trachea, while maintaining blood flow through the thoracodorsal artery. Indocyanine green near-infrared fluorescence angiography was visualized using a FLUM-808 multispectral fluorescence organoscope. Results. Sixty days after implantation, the tracheal cartilaginous framework macroscopically appeared to be intact in both animals, tightly integrated into the muscle tissue. The framework retained its natural color. After intravenous injection of indocyanine green, the tracheal vessels were visualized in both animals. Intercartilaginous vessels and portions of the cartilaginous semi-rings devoid of vessels were clearly distinguished. The entire implanted segment was almost uniformly vascularized. No local disruptions in blood supply were observed. The fluorescence brightness of the tracheal vessels was 193 ± 17 cu and 198 ± 10 cu in animals 1 and 2, respectively. The average muscle brightness in the implantation zone was 159 ± 9 cu and 116 ± 8 cu in animals 1 and 2, respectively. Conclusion. Indocyanine green fluorescence angiography is characterized by high-contrast images and high sensitivity. This facilitates vascular patency visualization and allows to assess the degree of neoangiogenesis after experimental transplantation of the tracheal segment, at different stages of experiment, without euthanizing the animal.
Objective: to evaluate the expression levels of miRNA (miR-27, miR-101, miR-142, miR-339 and miR-424) and its relationship with clinical and laboratory parameters in lung transplant recipients. Materials and methods. The study included 57 lung recipients aged 10 to 74 years (35 ± 15), including six children (9%) – four boys 10, 12, 13 and 17 years and girls 13 and 14 years old – and 51 adult recipients, including 30 men (62.5%). The control group was made up of 14 healthy individuals that were not significantly different by gender and age. Expression levels of the microRNAs studied in blood plasma were determined via quantitative polymerase chain reaction (PCR). Correlations of miRNA expression levels with complete blood count and biochemical blood test indicators were analyzed. Results. Patients with end-stage chronic respiratory failure (potential lung recipients) were found to have significantly higher expression levels of miR-27, miR-101 and miR-339 in plasma than the healthy individuals (p = 0.02, p = 0.03 and p = 0.01, respectively). The expression level of miR-339 correlated with the age of potential lung recipients (p = 0.04). It was a negative correlation (r = –0.46). The expression levels of the other four miRNAs were age independent. The average expression level of miR-424 in lung recipients in the long-term period after lung transplant was higher than in waitlisted patients (p = 0.03). Analysis of the relationship between miRNA expression levels and external respiration function in the long-term post-transplant period showed that miR-142 expression level (r = 0.61; p = 0.04) positively correlates with the Tiffeneau-Pinelli index. This strong correlation, which exceeds 85%, indicates the presence of restrictive lung diseases. A year and more after transplantation, it was found that in the recipients, there were close positive correlations between miR-27, miR-142, miR-424 expression levels and blood leukocyte concentration, as well as between the miR-142 expression level and the sCD40L concentration during this period. Conclusion. A comparative study of the expression level of miRNAs (miR-27, miR-101, miR-142, miR-339 and miR-424) in the blood plasma of patients suffering from end-stage chronic lung diseases of various origin and in lung recipients enables us to conclude that further studies of the miRNA panels are needed in order to assess their effectiveness as potential molecular and genetic markers of post-transplant complications.
Assessing the viability and monitoring the function of liver graft in the early postoperative period are critical clinical tasks. One possible solution is to determine the changes in concentration of blood glucose, its metabolites and glycerol in the graft using interstitial microdialysis. Objective: to study the dynamics of interstitial glucose, lactate, pyruvate and glycerol in the early post-liver transplant period – depending on the initial graft function (IGF) – and to compare with the results of standard laboratory blood tests. Materials and methods. Four selected clinical observations of deceased donor liver transplantation are presented. Two of the observations showed normal IGF, one observation – early allograft dysfunction (EAD), complicated by hepatic artery thrombosis (HAT), while one observation demonstrated primary non-function (PNF). Collection of microdialysis samples began after arterial reperfusion of the liver graft and continued continuously for 7 days or until death. Standard blood biochemistry and coagulation tests were performed at least once a day. Results. With normal IGF and a smooth postoperative period, interstitial concentrations of glucose, lactate, pyruvate and glycerol remained stable throughout the observation period, ranging from 5 to 20 mmol/L, 1.1 to 7.5 mmol/L, 90 to 380 μmol/L, and 10–100 μmol/L, respectively. EAD was associated with initially higher levels of glucose, lactate, and pyruvate. With HAT development, there was a rapid (within 2–4 hours) five-fold increase in interstitial concentration of lactate with simultaneous decrease in glucose and pyruvate levels to 0.1 mmol/L and 11 μmol/L, respectively. In the case of PNF, there was an initially high concentration of interstitial lactate – 16.4 mmol/L, which increased further to 35.5 mmol/L. Glucose concentration was close to 0. Changes in interstitial glucose, its metabolites and glycerol concentrations chronologically preceded the corresponding changes in peripheral blood composition by 3–5 hours. Conclusion. Microdialysis measurement of interstitial glucose, lactate, pyruvate and glycerol concentrations facilitates real-time monitoring of liver graft viability and function. The high sensitivity of the method could help in accelerating diagnosis of vascular complications (HAT in particular), as well as graft dysfunction with other causes. Therefore, the method is feasible in clinical practice.
Heart Transplantation and Assisted Circulation
Objective: to analyze the inflow cannula of an implantable axial-flow blood pump for a long-term left ventricular assist system in order to minimize thromboembolic complications. Materials and methods. Hemodynamics was considered for 4 different designs of the inflow cannula, from 0 mm to 25 mm long. Areas at the base of the cannula received the most attention. Analysis was performed using the OpenFOAM software. Results. It was revealed that sizes of stagnation and recirculation zones directly depended on the length of the cannula when placed in the left ventricle. Accordingly, longer cannula increases the risk of thrombosis. Conclusion. The design of an inflow cannula determines the likelihood of thrombosis in the cannula. Longer inflow cannula increases stagnation and recirculation zones. This provides a basis for a search for other possible modifications.
Objective: to carry out the implantation of an artificial left ventricle of the heart based on a disk-type pump in an acute experiment on a large mammal (mini-pig). Materials and methods. To test the surgical technique of implantation and assess the biocompatibility of the apparatus for mechanical support of blood circulation based on a viscous friction pump, an acute experiment was conducted on an animal. A large mammal (mini-pig weighing 90 kg) was used as an experimental model. The implantation of the pump was performed extracorporeally according to the scheme «the apex of the left ventricle – the descending thoracic aorta». During the experiment, invasive blood pressure, central venous pressure, cardiac arrhythmias, body temperature, blood gas composition, activated coagulation time were monitored. Under the control of transesophageal echocardiography, the pump operation mode was set with parameters – speed 2400–2600, productivity 4 ± 0.5 l/min, average IAD – 70–80 mm Hg. Results. In the course of the experiment, the fundamental possibility of using the developed disk-type pump as a device for supporting blood circulation was proved. For 4 hours, the pump provided adequate hemodynamic parameters with an average productivity of 4 ± 0.5 l/min and 2500 rpm. After 4 hours of operation of the pump in the conditions of inactivated heparin (AST – 114 sec), no blood clots were found between the pump disks. Conclusion The hemodynamics feature of the disk pump allows you to develop sufficient performance parameters to ensure adequate blood circulation. The mechanism of action of the «boundary layer» minimizes the risk of blood clots in the pump cavity. However, the topographic and anatomical features of the pig’s body do not allow experiments with a long observation period.
The aim of the study was an in vitro hydrodynamic study of the developed prosthetic heart valve of the second generation, designed to carry out an implantation using «valve-in-valve» method. Material and methods. Prototypes of the developed prosthesis were studied under simulated physiological conditions of the heart using a Vivitro Labs pulse duplicator (Canada) in a comparative aspect with «UniLine» clinical commercial aortic valve bioprosthesis (Russia). Samples were tested by simulating sutureless implantation procedure. Results. The developed valves showed satisfactory hydrodynamic characteristics – for all cases of «implantation» from the position of the average trans-prosthetic gradient (6.1–11.1 mm Hg) and the effective orifice area (1.60–1.81 cm2 ). The analysis of the regurgitation fraction allowed us to determine the optimal sizes for implantation using «valvein-valve» method, which subsequently will form the basis of sizing guidelines for size selection. A qualitative analysis of the leaflet’s work demonstrated the presence of slight asymmetry for a number of prostheses – in case of mismatch of sizes when simulating «valve-in-valve» procedure. Conclusion. The tests demonstrate the viability of the developed design from the standpoint of hydrodynamic efficiency and determines the basic rules of selecting a prosthesis for reimplantation relative to the primary valve.
Clinical Cases
Regenerative Medicine and Cell Technologies
Objective: to study the effect of various types of suture materials, potentially suitable for cardiovascular surgery, on experimental surgical outcomes. Materials and methods. Polypropylene sutures (Prolene 6/0), titanium nickelide (TiNi) sutures (6/0) and absorbable polydioxanone sutures (Monoplus 6/0) were used in the study. Male Wistar rats were used for in vivo studies. The effect of suture materials on abdominal adhesions was studied. In vivo calcification process was examined, and response of blood components in contact with suture materials was also assessed in vitro. Results. There is a negative inflammatory response to suture materials. The severity of this response depended on the type of material used. Polypropylene sutures demonstrated the most severe inflammatory response provoking massive adhesion formation. In addition, large calcium deposits were found both in the suture area and in the thickness of the biomaterial, stitched with prolene and implanted subcutaneously in the rats. Titanium nickelide sutures showed high hemocompatibility and biocompatibility. The Monoplus sutures caused minimal inflammatory response and provoked calcification of the biomaterial to a lesser degree. Conclusion. The suture material could have significant effects on surgical outcomes and could cause postoperative complications.
Organ preservation
LITERATURE REVIEWS
Transplantological Aid Management
Objective: to provide guidelines on how to improve the quality of health care in transplantation services for Russian regions; the recommendations were prepared by the Shumakov National Medical Research Center of Transplantology and Artificial Organs (Shumakov Center) based on field trips made by Shumakov Center from 2018 to 2019 in order to assess the status and development prospects of organ donation and organ transplantation in various regions of the Russian Federation. Materials and methods. Based on the over 40 field trips made by the Center, analytical reports, as well as recommendations were prepared by the Ministry of Health of Russia for regional health authorities and transplant centers. Guidelines were selected from these recommendations to address the most significant and common gaps and errors inherent in organ donation and organ transplantation in the regions. On the basis of the analytical reports, guidelines for the roadmap (plan) for organ donation and transplantation in the region were prepared. Results. A methodological framework for solving a wide range of managerial challenges on organ donation and organ transplantation is proposed for heads and experts at healthcare institutions and medical organizations. Conclusion. Organ donation and organ transplantation should be developed for the regions using the roadmap (plan) prepared and approved by the regional health authority. This approach brings stability and consistency to the development process, creates a resource base for development, and provides coordination and oversight.