Aim. To study the frequency of fatty hepatosis in liver biopsies of consecutive brain death donors before cold preservation. Materials and methods. Liver biopsies (before cold preservation) of 300 consecutive donors with brain death were studied. Histological preparations were stained with hematoxylin and eosin, and tricolor Masson staining was performed. Results. The frequency of different degrees of fat hepatosis in men and women did not differ significantly (>0.05). Fat dystrophy of hepatocytes was absent in more than half of the cases (n = 182; 60.7%). A slight degree of fatty degeneration was diagnosed in 57 (19,0%) donors. In total, 239 (79.7%) donor livers were absolutely suitable for transplantation. Moderate degree of steatosis, which is associated with early biliary complications, was detected in 18 (6.0%) cases, and severe degree, which is a contraindication to the use of the organ for transplantation, was detected in 43 (14.3%) cases. Conclusion. Before cold preservation, liver from brain death donors is relatively rarely unsuitable for transplantation.

The emergence of direct-acting antivirals (DAAs) has become the basis for a new potential treatment for chronic hepatitis C (CHC) in patients with decompensated cirrhosis, who previously had no other alternative than liver transplantation (LT). However, optimal timing of antiviral therapy (AVT) remains an issue. Objective: to present a spectrum of clinical outcomes in LT waitlisted patients with HCV-related cirrhosis, who received and did not receive DAA therapy. Materials and methods. Enrolled for the study were 49 waitlisted patients with HCV-related end-stage liver diseases. The patients were divided into 2 groups: Group 1 included 40 patients who received DAA therapy before LT, while Group 2 consisted of 9 patients who did not receive antiviral treatment while on the LT waiting list. Results. The sample was represented in most cases by patients who had MELD/Na score <20. Only six had MELD/Na score >20, but <25. At the time of analysis, 38 patients had reached 12 weeks post AVT. Of these, 35 (92.1%) had sustained virologic response (SVR). Of these, 51.4% (n = 18) of cases showed decreased MELD/Na. There were no changes in 22.9% (n = 8). Increased MELD/Na was noted in 25.7% (n = 9). In 42.8% (n = 15) of cases, sustained elimination of HCV infection led to delisting. Among patients without SVR, increased MELD/Na was observed in all cases (n = 3). In the non-AVT group, one patient showed improved liver function (11.1%); in the rest, MELD/Na either remained stable or continued to increase - 44.5% (n = 4). A comparison of the frequency of deaths depending on AVT showed statistically significant differences (p < 0.001, V = 0.728). Among the non-AVT patients, the likelihood of waitlist death increased 66.5 times (95% CI: 7.99-554). Conclusion: DAA therapy carries significant advantages for waitlisted patients with MELD/Na score <25.

Objective: to evaluate the effectiveness of vacuum-assisted closure (VAC) therapy in comparison with standard treatments for infected and chronic non-healing wounds after kidney transplantation. Materials and methods. From June 2018 to November 2019, 75 kidney transplants from deceased donors were performed at the Transplantation Ward of Botkin City Clinical Hospital. There were 47 men (62.6%) and 28 women (37.4%). Standard surgical technique was used. Immunosuppressive therapy was carried out according to a three-component scheme with anti-CD25 monoclonal antibody induction (basiliximab) intraoperatively and on day 4. All patients received antibiotic therapy with protected third-generation cephalosporins for 7 days after surgery. Postoperative complications were evaluated according to the Clavien-Dindo classification. Standard methods, including daily dressings using modern dressing materials (group I) and VAC therapy (group II) were used for treating infected and chronic non-healing wounds. Results. 30-day mortality in the postoperative period was zero. Postoperative complications were recorded in 11 patients (14.6%), of which 7 had postoperative wound complications. Group I included 3 patients (1 with a Klebsiella pneumonia-infected wound and 2 with chronic non-healing wounds and no microflora growth). Group 2 had 4 patients (3 with infected wounds (Esherichia coli - 1, Klebsiella pneumonia - 2) and 1 with chronic non-healing wound). Complete cleansing of wound, absence of bacterial growth according to the microbiological examination, and maturation of granulations according to histological examination were considered as the criteria upon which a wound could be sutured in both groups of patients. The average time between the start of treatment and secondary suturing in group 1 patients was 33.11 ± 5.43 (28-37) and 15.01 ± 3.15 (13-17) days in group 1 and group 2 respectively. Conclusion. VAC therapy in patients with wound complications resulting from kidney transplantation, in comparison with standard treatment, can achieve rapid wound cleansing, acute inflammation relief and accelerated maturation of mature granulation tissue, thereby improving treatment outcomes in this category of patients.

Aim: to study the effect of cardiac contractility modulation (CCM) in patients with chronic heart failure (CHF) and atrial fibrillation (AF). Materials and methods. In a group of 100 patients with CHF and AF, the following studies were performed before implantation of the CCM and after 6 months of follow-up: 12-channel ECG, transthoracic Echocardiography, 6-minute walk test, determination of the level of pro-natriuretic N-terminal peptide (NT-proBNP), and a questionnaire based on the Minnesota quality of life questionnaire for patients with CHF (MHFLQ). All patients received long-term optimal medication therapy for CHF before surgery. Results. The results show a positive effect of the use of MCC in patients with CHF and AF on reverse LV remodeling, functional class of CHF, and levels of NT-pro-BNP regardless of the form of AF. Conclusion. The use of MCC may be a promising treatment method in addition to optimal medication therapy in patients with CHF and AF.

A young female patient who developed anastomotic biliary stricture following an orthotopic liver transplantation was observed. A self-expandable metallic stent was placed to correct the stricture. At the 8th month of her repeat pregnancy, the stent broke asymptomatically into half. Fortunately, the second childbirth, like the first one, had no complications. Eighteen months later, due to obstruction of fragments by sludge and gallstones, re-stenting was performed with a coated biliary stent. Four years and five months later, recurrent jaundice occurred due to occlusion of the second stent. This was addressed by surgical removal of both stents. Two years after surgery, the bile ducts remain completely patent. We found only two cases in literature on a similar extremely rare biliary stenting complication. It has been suggested that stent deformation may be related to pregnancy. The feasibility of using stenting in benign biliary strictures in some clinical situations is discussed.

Objective: to study the peculiarities of the induction effect of total RNA (tRNA) from xenogenic bone marrow cells (BMCs) on regeneration processes in the recipient's native liver with extensive liver resection using an adoptive transfer model. Materials and methods. The study was carried out on an adoptive transfer model using male Wistar rats (n = 20) and guinea pigs (n = 17). The donors were rats (n = 10). 12 hours after extensive liver resection (70-75%), tRNA was isolated from BMCs and injected into intact (non-operated) recipients intraperitoneally at a dose of 30 μg/100 g of weight. The induction effect of the tRNA on operated rats was studied in 3 groups of recipients: Group 1 (control, n = 5) - administration of saline to guinea pigs; Group 2 (control, n = 10) - administration of tRNA from a donor rat to a recipient rat (allogeneic transfer); Group 3 (experiment, n = 12) - administration of tRNA from a donor rat to a recipient guinea pig (xenogeneic transfer). In histological preparations of recipient livers, after 48, 72 hours and 7 days, we studied the mitotic activity of hepatocytes and the features of the microscopic picture of the liver. The significance of differences in the compared groups was assessed using the parametric Student's t-test. Results. The ability of BMC tRNA to tissue-specifically activate regenerative and immune responses in the liver after extensive resection was found to depend on the donor and recipient species identity. Introduction of allogeneic donor tRNA in the recipient's liver resulted in predominant enhancement in hepatocyte mitotic activity (p < 0.05). The use of xenogeneic donor tRNA leads to enhanced activity of only immuno-inflammatory reactions in the recipient's liver, such as sinusoidal cell activation, lymphocytic infiltration into sinusoids, and portal tract infiltration by inflammatory cells. Conclusion. To induce regenerative processes in the liver, tRNA obtained from allogeneic BMCs should be used.

Correcting the pleural cavity space or filling large residual cavities (up to 500-700 cm³), arising as a result of extensive combined resections of the lung or extrapleural pneumolysis in tuberculosis and other lung diseases, still remains a challenging issue. The surgical methods used to correct the pleural cavity space are traumatic in nature. Moreover, various biological and synthetic materials used are not effective enough. Objective: to conduct an in vivo study of the biocompatible properties of laboratory samples of porous materials based on polylactide (PLA) and polycaprolactone (PCL) as potential materials for pleural implants development, as part of the general problem of developing a resorbable porous implant for intra- and extrapleural implantation and in situ formation of a «biological filling» to correct the volume of the pleural cavity. Materials and methods. In vivo subcutaneous implantation was performed in Wistar rats. The experiment involved the following samples: No. 1 - 3.0%; No. 2 - 4.0%; No. 3 - 1.7%. The ratio of the polymers in the solution was, respectively: 3/1, 1/3 and 1/1 PLA/PCL. Highly porous implants were obtained by lyophilization. The porosity of the samples ranged from 96.0% to 98.3%. The Young's modulus was from 100 to 1800 kPa. In the control group, a Mentor silicone implant shell was used. The explantation time was 1, 2, 3, 4, 5, 8, 12, 14 weeks. Histological, histochemical and immunohistochemical studies of explants and surrounding local tissues were conducted. Results. Reaction of local tissues to the implantation of three types of samples of different composition from PLA/PCL, accompanied by material resorption processes, replacement by fibrous tissue, vascularization and encapsulation, without perifocal inflammation and reactive changes, indicates the biocompatibility of the materials studied. In control samples with silicone implant, a long-lasting perifocal reaction from eosinophilic leukocytes was revealed, which prevents us from excluding the possibility of an allergic reaction to the implant material in the surrounding tissues. Conclusion. In vivo experiments on the small animals show the biosafety and high biocompatibility of laboratory samples of bioresorbable highly porous matrices based on polylactide and polycaprolatcon as potential materials for development of pleural implants. Further studies with scaling of laboratory samples and a detailed study of the dynamics of biodegradation of porous matrices in vivo in large animals are required. The need for further improvement in laboratory samples of bioresorbable pleural implants is associated with giving the porous matrices antibacterial, bioactive and X-ray contrast properties.

3D Bioprinting is a dynamically developing technology for tissue engineering and regenerative medicine. The main advantage of this technique is its ability to reproduce a given scaffold geometry and structure both in terms of the shape of the tissue-engineered construct and the distribution of its components. The key factor in bioprinting is bio ink, a cell-laden biocompatible material that mimics extracellular matrix. To meet all the requirements, the bio ink must include not only the main material, but also other components ensuring cell proliferation, differentiation and scaffold performance as a whole. The purpose of this review is to describe the most common materials applicable in bioprinting, consider their properties, prospects and limitations in cartilage restoration.

Calcification of biomaterials used in prosthetic heart valves has been a challenging issue in cardiovascular surgery. The objective of this work is to compare the efficiency of polyvinyl alcohol (PVA) and tannic acid (TA) modification of xenomaterials, pre-stabilized with glutaraldehyde (GA) and ethylene glycol diglycidyl ether (EGDE), in reducing calcification. Analysis of mechanical properties evaluated under uniaxial tension, showed a significant increase in the tensile strength of the test samples compared to the control (unmodified) samples (p < 0.05). Additional treatment of GA-fixed tissue with PVA and TA significantly reduced the amount of calcium in the samples implanted into rats for a 60-day follow-up (p < 0.05). The level of calcification of samples prestabilized with EGDE and treated with PVA and TA did not differ from the control group (p = 0.063). Cumulative analysis of the study results demonstrated that the GA-fixed biomaterial modified with PVA and TA can reduce calcium-binding activity and increase strength. This indicates the prospects for clinical application of the proposed treatment methods. This being said, the issue of long-term body response requires further study of the long-term stability of the modified biomaterial under physiologic blood flow conditions.

This paper presents the immediate outcomes of valve-sparing operations on the aortic valve and ascending aorta in radical correction of congenital and acquired heart disease. Materials and methods. The study enrolled 50 patients with aortic insufficiency who were operated upon at Shumakov National Medical Research Center of Transplantology and Artificial Organs from 2011 to 2019. The mean age was 48 ± 16 years, 64% of them were men (n = 32). The study included patients with tricuspid (n = 36, 72%) and bicuspid (n = 14, 28%) aortic valves. Aortic valve reimplantation was performed in 32 (64%) patients, aortic root remodeling - in 1 (2%). 17 (34%) patients had no aortic root reconstruction or remodeling. Aortic valve reimplantation was done in 4 (8%) cases in combination with coronary artery bypass grafting, and in 4 (8%) with mitral and tricuspid valve repair. Results. Thirty-day mortality was 0%. In 1 case (2%), a permanent pacemaker was installed due to complete atrioventricular block. There were no neurological and coronary events, and cases of endocarditis. In all patients (100%), aortic valve insufficiency after surgical correction did not exceed grade 1 according to echocardiographic follow-up examination. On aortic valve mean and peak gradients were 8 ± 6 and 15 ± 7 mm Hg, respectively. Findings. Type I and II valve-sparing reconstructive surgery (for bicuspid and tricuspid aortic valves) is an excellent alternative to prosthetic repair with great postoperative outcomes, low valve-associated complications and low mortality. 

In 2017, the European Society of Cardiology outlined the importance of the problem of diagnosing myocardial ischemia-reperfusion injury following coronary artery bypass grafting. Myocardial injury can be accompanied by a critical decline in the cardiac index and an increase in cardiac troponin I plasma levels. The prognostic value troponin I elevation after coronary artery bypass grafting is poorly understood. Objective: to determine the prognostic value of troponin I plasma levels in relation to a fall in the cardiac index after coronary artery bypass grafting (CABG). Task: To determine the probability the cardiac index falling below 2.2 for troponin I levels in the first hours, and on days 1, 2, 3, 4 after CABG. Materials and methods. The single-center, non-randomized prospective study, running from 2016 to 2019, included 336 patients admitted for elective surgical treatment of coronary artery disease. The CABG patients were divided into three observation groups: off-pump (n = 175), on-pump (n = 128), and pump-assisted (n = 33). Troponin I levels were measured in the first hours, and on days 1, 2, 3, 4 after surgery using the Pathfast Compact immunoassay analyzer. Cardiac index was measured by invasive method. Results. In patients with a cardiac index higher than 2.2, troponin I level did not exceed 0.5 ng/mL in the off-pump group, 6 ng/mL in the on-pump group, and 3.5 ng/mL in the pump-assisted group. Patients with cardiac index lower than 2.2 have comparable troponin I levels in all groups - 21 ng/mL. Troponin I thresholds on day 1 after surgery, which, when exceeded, was associated with the likelihood of the cardiac index falling below 2.2, was 3.78 ng/mL in the off-pump group, 9.67 ng/mL in the on-pump group and 17.06 ng/mL in the pump-assisted group. Conclusion. After off-pump CABG, clinically significant myocardial injury should be expected at lower troponin I levels (3.78 ng/mL) than after on-pump CABG (9.67 ng/mL) and pump-assisted CABG (14.7 ng/mL).

Re-interventions after pulmonary autograft aortic valve replacement (Ross procedure) may be associated with dysfunction of the neoaortic, neopulmonary, or both operated valves. Late dysfunction, other than infective endocarditis, is associated with underlying conditions, technical errors, and unsuitable pulmonary trunk replacement materials. Re-interventions are technically complex, while tactical approaches have not been definitively formulated. Objective: to analyze re-interventions in patients after Ross procedure, technical approaches and immediate outcomes. Material and methods. Between 2001 and 2019, 14 patients were reoperated upon within 2 days to 21 years after primary Ross procedure. Early prosthetic endocarditis (2) and technical errors (1) were the reasons for early postoperative re-intervention. Neoaortic valve insufficiency (7), including pulmonary valve dysfunction (2), pulmonary valve degeneration (2), pulmonary prosthetic valve endocarditis (1), aortic, pulmonary and mitral valve endocarditis (1) were the reasons for late postoperative re-intervention. Based on the lesion volume, neoaortic valve replacement (3), neoaortic root replacement (6), including pulmonary valve/trunk replacement (8), and pulmonary trunk stenting (2) were performed. Results. In-hospital mortality was 7.1%. One patient died of early endocarditis after primary procedure. The postoperative period for the remaining patients was uneventful. Microscopic examination of the neoaorta revealed fragmentation of elastic fibers and rearrangement of tissue histoarchitectonics. In the pulmonary position, the aortic allograft and stentless xenograft had severe calcification and valve stenosis. Conclusions. Neoaortic valve insufficiency associated with cusp prolapse and neoaortic root dilatation may be the reasons for re-interventions after the Ross procedure. The second reason for re-interventions is valve graft dysfunction in the pulmonary trunk position. Elective reoperations on the neoaortic root and/or lung graft, despite the large volume, can be performed with low mortality and morbidity. Aortic allografts and xenografts for reconstruction of the right ventricular outflow tract (RVOT) is unjustified due to early and more severe dysfunction compared to pulmonary allograft.

Objective: to analyze the in-hospital and long-term outcomes of classical carotid endarterectomy (CEE) in extended atherosclerotic lesions in comparison with the outcomes of this operation in local atherosclerotic plaque (AP). Materials and Methods. This study, which lasted from January 2010 to December 2020, included 148 patients with extended AP and hemodynamically significant internal carotid artery (ICA) stenosis. The term “extended” was understood as a hemodynamically significant lesion ≥ 5 cm long. These patients made up Group 1. Group 2 was formed over the same period of time from 632 patients with hemodynamically significant stenosis <5 cm long. In both cohorts, CEE with repair of the reconstruction zone with a diepoxide-treated xenopericardial patch was performed. Long-term follow-up was 71.4 ± 45.6 months. Results. The groups were comparable in terms of frequency of in-hospital complications: death (group 1: 0.67%, n = 1; group 2: 0.5%, n = 3; p = 0.74; OR = 1.42; 95% Cl 0.14-13.6), myocardial infarction (MI) (group 1: 0.67%, n = 1; group 2: 0.5%, n = 3; p = 0.74; OR = 1.42; 95% CI 0.14-13.6), ischemic stroke (group 1: 0%; group 2: 0.5%, n = 3; p = 0.91; OR = 0.6; 95% CI 0.03-11.8), combined endpoint (death + MI + stroke) (group 1: 1.35%, n = 2; group 2: 1.4%, n = 9; p = 0.74; OR = 0.94; 95% CI 0.2-4.43). The groups were also comparable in terms of frequency of long-term complications: death (group 1: 2.0%, n = 3; group 2: 2.05%, n = 13; p = 0.76; OR = 0.98; 95% CI 0.27-3.5), MI (group 1: 2.7%, n = 4; group 2: 2.4%, n = 15; p = 0.95; OR = 1.14; 95% CI 0.37-3.49), ischemic stroke (group 1: 5.4%, n = 8; group 2: 5.2%, n = 33; p = 0.9; OR = 1.03; 95% CI 0.46-2.29), ICA occlusion and restenosis (group 1: 12.8%, n = 19; group 2: 13.3%, n = 84; p = 0.99; OR = 0.96; 95% CI 0.56-1.63), combined endpoint (death + MI + stroke) (group 1: 10.1%, n = 15; group 2: 9.6%, n = 61; p = 0.98; OR = 1.05; 95% CI 0.58-1.91). Analysis of survival graphs revealed no significant intergroup differences for all types of complications (lethal outcome: p = 0.56; MI: p = 0.73; stroke/mini-stroke: p = 0.89; ICA restenosis/occlusion: p = 0.82; combined end point: p = 0.71). Their increase was uniform in both groups. However, more than half of all ICA restenoses and occlusions were visualized in the first 6 months after CEE. Conclusion. Implantation of a long patch (≥ 5 cm) is not characterized by increased incidence of restenosis and all adverse cardiovascular events during in-hospital and long-term follow-up.

The risk of severe infectious complications associated with provision of medical care continues to be a pressing issue in modern surgery. Legionella pneumophila, characterized by its wide distribution in water supply systems and is highly active in film formation, represents a dangerous/important cause of hospital-acquired pneumonia. Patients requiring immunosuppression, including organ transplant recipients, are in the special risk group. Prevention of hospital-acquired legionellosis in patients at risk is essential due to severe clinical manifestations and high mortality. Objective: to summarize the practical experience in detecting contamination of water supply systems by Legionella pneumophila strains in multidisciplinary hospitals in Moscow. Materials and methods. Isolation of Legionella pneumophila strains from water and biofilms of water supply systems in multidisciplinary hospitals in Moscow and serotyping of this pathogen using bacteriological, molecular genetic and enzyme immunoassay methods. Results. Legionella pneumophila content in water reached high levels. The peculiarities of Legionella pneumophila contamination of hot water supply systems included formation of stable biofilms, in which other hospital-acquired pathogens were also identified. The share of Legionella pneumophila «SG 1», which causes up to 80% legionellosis cases in the world, was 13% in the water of the hospitals surveyed. The most effective measures for prevention of legionellosis are actions aimed at ensuring water biosecurity. Conclusion. There are potential risks of disease in the surgical wards of hospitals providing medical care, including in immunocompromised patients. Due to potential risks, prevention of hospital-acquired legionellosis is a necessary component of ensuring the safety of treatment for immunosuppressed patients.

Heart transplantation continues to be the gold standard treatment for end-stage chronic heart failure. As with any cardiac surgery, heart transplantation is associated with postoperative complications. One of the most common complications is postoperative pericardial effusion. Heart recipients have a greater risk of developing pericardial effusion than patients after cardiac surgery on their own heart, due to surgical and immunological features. Severe pericardial effusions negatively affect the postoperative period and may be the cause of life-threatening conditions. Identification of risk factors, prevention, early diagnosis and treatment of this disease can significantly reduce the risks of adverse events in this group of patients. The purpose of this literature review is to analyze the development and course of pericardial effusion in heart recipients in world practice.

Bronchial complications are among the main causes of impairing postoperative period and thansplant failure. Severe bronchial complications are very rare but have a high mortality rate. Light forms decrease transplant function and while progressing can leads to life-threatening conditions without required treatment. Nowadays there is a huge necessity in classification of diagnostic and bronchial complications treatment on different terms after lung transplantation. Methods of observation bronchoscopy and interventional bronchology are allowing us to realize prevention, diagnostic and treatment bronchial complications.

Tissue engineering has significant potential for solving the problems of durability of biological tissues when used in cardiac and vascular reconstructive surgery. A decellularization technology has been proposed for obtaining a biomaterial, morphologically and functionally similar to the damaged human heart tissue. This review discusses various aspects and models of biological tissue decellularization, including the modern technology of using supercritical carbon dioxide as the most eco-friendly and promising method.

This paper reviews the current main approaches to valve-sparing aortic root reconstruction. The advantages of valve-sparing surgeries are obvious - low mortality, longer survival, better quality of life of the operated patients, since the techniques save the heart's pumping reserves and free the patient from continuous intake of direct-acting oral anticoagulants and laboratory control of the hemostasis system, as well as other prosthesis-associated specific complications.

Primary biliary cholangitis (PBC), formerly known as primary biliary cirrhosis, is an organ-specific autoimmune disease predominantly affecting middle-aged women. It does not occur in children. PBC prevalence varies depending on the geographic location of the country. Over the past 30 years, there has been an increased incidence of PBC, while significant progress has been made in understanding the pathogenesis of PBC due to the development of innovative technologies in molecular biology, immunology and genetics. The presence of antimitochondrial antibodies and cholestasis on biochemical analysis is sufficient to make a diagnosis, without the need for liver biopsy. Small- and medium-sized bile ducts are the targets of PBC. In the first stage of the disease, granulomatous destruction of the bile ducts occurs; in the second stage, loss of bile ducts, their proliferation, increased size of the portal tracts with chronic inflammation; in the third stage - fibrosis with septal formation, loss of bile ducts and cholestasis; in the fourth stage - liver cirrhosis. Previously, the survival rate of PBC patients ranged from 7.5 to 16 years. However, it has improved significantly with ursodeoxycholic acid and obeticholic acid treatment. If there is no effect from treatment and end-stage liver failure sets in, liver transplantation is performed.

According to the World Health Organization, corneal blindness is the fourth most common cause of blindness and visual impairment worldwide. In Russia, up to 18% of blindness is caused by corneal damage. Limbal stem cell deficiency (LSCD) is one of the causes of corneal blindness and visual impairment due to anterior epithelial replacement with fibrovascular pannus. Bilateral LSCD may develop in patients with aniridia, Steven-Jones syndrome, and severe corneal burns of both eyes, leading to severe decrease in visual acuity in both eyes and, as a consequence, physical disability associated with blindness. In such cases, cell therapy, based on autologous oral epithelial culture as an alternative to allogeneic limbus transplants, is proposed for reconstruction of the anterior corneal epithelium. This new treatment method promotes corneal reepithelization, better visual acuity, reduced nonspecific ocular complaints and improved quality of life of patients. The effectiveness and significant increase in the frequency of transparent engraftment of donor corneas after cell therapy drives huge interest in this topic all over the world. This review presents literature data on the features of histotopography and methods for obtaining a cultured autologous oral mucosal epithelium, on cell markers that are used to identify epithelial cells, and on methods for creating cell grafts for subsequent transplantation to the corneal surface in LSCD patients.