Corneal fibroblasts and keratocytes differentiated from fibroblasts were used for cell therapy. Mechanical injury to the mouse cornea was modeled in the form of a tunnel defect into the depth of the cornea in the central optical zone. Corneal thickness and transparency were assessed by optical computed tomography at 2, 4, 6, and 8 weeks after the injury. The therapeutic effectiveness of fibroblasts was delayed and was associated with paracrine influences of keratocytes. The conditioned medium of keratocytes reduced adhesion ability of fibroblasts and their proliferative and migratory activities. In case of keratocyte therapy, corneal thickness and transparency began to recover after 2 weeks and reached the level of a healthy cornea by week 8. The results of our study demonstrate the efficacy of correction of corneal opacification by stromal cells.

Polymorphism of genes of transforming growth factor TGFB and its receptors (TGFBRI, TGFBRII, and TGFBRIIII) in patients with primary open-angle glaucoma was analyzed. The frequency of the TGFBRII CC genotype in patients is increased relative to the control group (OR=6.10, p=0.0028). Heterozygosity in this polymorphic position is reduced (OR=0.18, p=0.0052). As the effects of TGF-β is mediated through its receptors, we analyzed complex of polymorphic variants of the studied loci in the genome of patients. Two protective complexes consisting only of receptor genes were identified: TGFBRI TT:TGFBRII CG (OR=0.10, p=0.02) and TGFBRII CG:TGFBRIII CG (OR=0.09, p=0.01). The study showed an association of TGFBRII polymorphism with primary open-angle glaucoma and the need to study functionally related genes in the development of the disease, which should contribute to its early diagnosis and prevention.

Relevance. Around the world, corneal blindness caused by impaired transparency is one of the leading causes of vision loss. Damage to the cornea can also lead to the development of opacities. The pathogenesis of corneal opacities is based on pathomorphological changes in the corneal stroma. To explore the therapeutic potential of biomedical products and develop new treatments, a standardized and reproducible in vivomodel of corneal stromal injury is needed. In experimental ophthalmology, mouse models of various corneal injuries are widely used, but the dynamics of posttraumatic restoration of the ultrastructural organization of the stroma have been little studied. Purpose. To evaluate structural and functional changes in the corneal stroma of mice in the dynamics of post-traumatic recovery. Material and methods. Injury to the corneal stroma was performed using a sharp 33G needle in the form of a tunnel defect in the thickness of the cornea of C57BL/6 mice. The presence of post-traumatic damage and the dynamics of corneal recovery were assessed using instrumental methods (biomicroscopy of the anterior segment of the eye with a fluorescein test, optical coherence tomography (OCT) before, immediately after the injury, and after 2 and 4 weeks after injury. Transmission electron microscopy was performed using a JEM 1400 electron microscope. Morphometry of digital images was performed using Image J software, calculating the minimum nearest interfibrillar distance, diameter and fibril density. Results. The formation of a corneal stromal wound was confirmed by the development of reactive edema according to OCT data and the accumulation of fluorescein in all damaged corneas. 2 weeks after the injury, complete epithelization of the wound surface was observed. According to OCT data, 2 weeks after the injury, an increase in corneal thickness by 34% and an increase in the opacity index by 1.5 times were recorded compared to a healthy cornea. By the 4th week of observation, the corneal thickness and opacity index remained at a high level in the injured cornea. After the formation of injury to the corneal stroma, a decrease in the diameter of collagen fibrils, an increase in fibril density and the shortest nearest interfibrillar distance were revealed throughout the observation period. onclusion. A mechanical model of corneal trauma performed using a sharp 33-gauge needle induces the formation of stable stromal opacities. Instrumental and morphological research methods did not reveal stromal restoration during the entire observation period (4 weeks). Key words: corneal trauma, optical coherence tomography, corneal thickness, corneal transparency, corneal morphology, corneal fibrils

Purpose. Description of a clinical case of a patient with the development of macular edema after suffering COVID-19. Material and methods. A description of the clinical observation of a patient with the development of macular edema after a new coronavirus infection (COVID-19) was carried out. Patient Ch., 68 years old, used a standard ophthalmological examination, ultrasound, optical coherence tomography. Results. The patient developed uveitis complicated by macular edema after a novel coronavirus infection (COVID-19). Antiinflammatory treatment was carried out using autologous platelet lysate in the conditions of the Novosibirsk branch of the S. Fyodorov Eye Microsurgery Federal State Institution. The prescribed treatment made it possible to maintain visual functions and improve life quality of the patient. Conclusion. COVID-19 is a serious pathology that can be accompanied by the development of panuveitis with cystic macular edema, which causes visual impairment. The management of such patients requires a multidisciplinary approach, regular monitoring and timely adequate treatment Keywords: uveitis; macular edema; new coronavirus infection

Aim. To identify endogenous risk factors for the development of glaucoma and cataracts based on the results of a comparative analysis of the nature of complex genetic trait distribution, including variants of genes for a number of cytokines and receptors for them, metalloproteinases, and their tissue inhibitors included in the genome of patients. Materials and methods. The study included 501 people of the Caucasian race born and living in the Siberian region of Russia. They were divided into three groups of patients – patients with primary open-angle glaucoma (POAG) (n = 99), patients with senile cataract (n = 100), and the control group (n = 302) without ophthalmic pathology. Genotyping of the analyzed polymorphic loci was carried out by real-time PCR using the SYBRGreen I dye and TaqMan probes and by restriction fragment length polymorphism (RFLP) for different polymorphisms. Results. The results of the study on the frequency of the analyzed genetic traits among patients with POAG compared to the control group showed the presence of combined genetic traits. The frequency of their detection in POAG was high and characterized by the two-digit value of the odds ratio, high values of specificity (99–100%), and high diagnostic coefficient. A direct comparison of the distribution of two ensembles of genes which protein products are involved in the extracellular matrix remodeling revealed a significant number of genetic traits characteristic of both diseases. This indicates significant differences in the implementation of the genetic predisposition to their development. Conclusion. The data obtained indicate the possibility of developing reliable laboratory criteria (riskometers) for predicting predisposition to the development of POAG and early diagnosis at the stage of preclinical manifestations.

Purpose. Purpose: to study the content of pro-inflammatory cytokines and matrix metalloproteinases in the intraocular fluid in patients with advanced stage of primary open-angle glaucoma. Material and methods. 50 patients were examined with a verified diagnosis of the advanced stage of poag. the control group consisted of 30 patients with a diagnosis of uncomplicated cataract. the concentration of the determined biologically active molecules was carried out by the method of flow fluorimetry on a two-beam laser analyzer -Bio-plex 200, Bio-rad, USA. Results. A statistically significant increase in the concentrations of Il-6, 8, 12, 17, mIp-1β matrix metalloproteinase-2 was found in the intraocular fluid of patients with poag. Conclusion. The data obtained indicate the significance of the local inflammatory process, as well as disturbances in the structure of the extracellular matrix in the mechanisms of poag development. Keywords: primary open-angle glaucoma, intraocular fluid, cytokines, matrix metalloproteinases

КЛИНИЧЕСКИЙ СЛУЧАЙ СЕМЕЙНОЙ ФОРМЫ НЕВОИДНОЙ ПИГМЕНТАЦИИ СЕТЧАТКИБратко Г. В. 1 , Дмитриев Д.Д. 1 , Трунов А.Н

Abnormal expression of matrix metalloproteinases (MMP) in watery moisture in patients with glaucoma may affect regulation of intraocular pressure (IOP). MMP activity is regulated by tissue metalloproteinase inhibitors (TIMP). The imbalance between tissue metalloproteinase inhibitors and matrix metalloproteinases may contribute to the development of glaucoma. Genetic factors, including polymorphism of matrix metalloproteinase genes and their inhibitors genes, can regulate the level of their expression, thereby affecting susceptibility to disease. Our aim was to perform comprehensive analysis of the MMP2 (rs243865), MMP3 (rs3025058), MMP9 (rs3918242) polymorphisms, and TIMP1 (rs4898), TIMP2 (rs8179090) tissue inhibitor genes polymorphisms in the patients with stage II (advanced) primary open-angle glaucoma. 99 patients (52 men and 47 women) with a verified diagnosis of stage II primary open-angle glaucoma were examined. The comparison group consisted of 100 age-matched persons (81 women and 19 men) without ophthalmic disorders. The single-nucleotide polymorphisms in promoter regions of MMP2, TIMP1, TIMP2 genes were analyzed by the TaqMan method, the MMP3 and MMP9 genes, by means of restriction fragment length polymorphism technique. Statistical evaluation was carried out using the specialized package of IBM SPSS Statistics 23 programs. The critical level of significance was assumed to be 0.05. The differences in the distribution of MMP2 rs243865 allelotypes with decreased frequency of TT genotype were found in the patient group and, vice versa, increased heterozygosity rates were revealed among them. In addition, the frequency of TIMP1 rs4898 heterozygous genotype was decreased in this group as compared to control sample. Four MMP/TIMP complex genotypes are positively associated with the development of pathology. Two of them were of bilocus type, i.e., MMP2-1306TC:TIMP2-418GG, and MMP3-11715A6A:TIMP1 372CC whereas two three-locus constellations were revealed, i.e., MMP2-1306TC:MMP9-1562CC:TIMP2- 418GG, and MMP3-11715A6A:MMP9-1562CC:TIMP1 372CC. There are nine MMP/TIMP complexes, the frequency of which in patients with glaucoma was significantly reduced when compared with control group. Polymorphism of regulatory regions of MMP2, MMP3, MMP9 genes and distinct gene variants of their inhibitors (TIMP1, TIMP2 genes) can be considered potential markers of the POAG development associated with an imbalance of MMP/TIMP activities.

ПОСТКОВИДНЫЕ ОФТАЛЬМОЛОГИЧЕСКИЕ ПРОЯВЛЕНИЯ НА ЗАДНЕМ ОТРЕЗКЕ ГЛАЗАБеляева А.И., Сафронова М

The proliferative activity of populations of stromal cells (fibroblasts) obtained from human corneal lenticles under conditions of their differentiation into keratocytes was studied. It was shown that during differentiation, the number of dividing fibroblasts and the frequency of divisions, and motor activity of these cells (speed of movement along the cell trajectory and the length of the trajectory) sharply decreased. These findings indicate a decrease in the proliferative activity of fibroblasts under conditions of their differentiation and transformation into keratocytes. A period of 17 days is sufficient for differentiation of corneal fibroblasts into keratocytes.

The article presents a literature review of the modern concept of anatomical and physiological structure and functioning of the cornea. The strict morphological structure and corneal tissue homeostasis ensure its transparency. Studying the mechanisms that regulate the constancy of the corneal tissue internal environment allows us to get closer to understanding the prospects forregenerative therapy for the corneal stroma pathology. The article discusses in detail the role and functional potential of corneal stromal cells, which are capable of reverse cytologic differentiation, which primarily ensures the maintenance of tissue homeostasis and corneal transparency. The functional activity of corneal cells can change for a number of reasons, which may be exogenous, iatrogenic (trauma, infection, etc.) or endogenous. Endogenous causes include: cell autoregulation pathologies (for example, enzyme defects); defects in transport systems leading to tissue hypoxia; disorders of the neuro-humoral regulation of trophism. The physical reason forthe violation of the corneal transparency is an increase in the light scattering. The article presents five main causes of increased light scattering in the opaque cornea, and also provides an overview of the main substances – components and products of cellular synthesis of corneal stromal cells: cytokines and growth factors (complex of the signal molecule and the SDF1/CXCR4 receptor, insulin-like growth factor 1, tumor necrosis factor alpha, intercellular adhesion molecule 1, erythropoietin, neurotrophic factors, etc.). Thus, corneal opacity can be caused by a single pathogenic mechanism or be the result of a complex effect of several factors. The main processes of tissue homeostasis regulation are aimed at maintaining the unique morphological structure of the cornea.

Immunohistochemical and ultrastructural analysis revealed signs of structural alterations in neurons and autophagy in all layers of the human retina at the end-stage glaucoma. The most pronounced destructive changes associated with swelling and destruction of mitochondria, endoplasmic reticulum, and Golgi apparatus, as well as structural signs of impaired synaptic activity and apoptosis were noted in ganglion, bipolar, and amacrine neurons. In the structure of photoreceptor cells, alone with destructive processes associated with structural alterations of rods and cones in the outer membrane discs, as well as swelling of organelles, we observed processes aimed at the maintenance of cell homeostasis. Structural signs of autophagy (mainly mitophagy) and changes of the ultrastructural organization in rod neurons were more pronounced than in cones.

Purpose: to study the structural organization of the vascular bed of human retina in the terminal stage of primary open-angle glaucoma (POAG).Material and methods. We performed a comparative immunohistochemical analysis of the content of vessels in the retina of 13 eyes of patients in the terminal stage of POAG, enucleated for medical reasons, and 17 eyes with uveal melanoma, using the markers of blood vessels endothelium CD34. The ultrastructural organization of the interstitium and endothelial cells of retinal microvessels was studied by electron microscopy and morphometry.Results. A significant increase in the volume density of the interstitium and a decrease in the volume density of CD34+-blood vessels in the retina of patients in the terminal stage of POAG, as compared with uveal melanoma, were revealed. An increased volume density of luminal and basal caveolae and the formation of transendothelial channels in the cytoplasm of endotheliocytes of retinal blood capillaries in the terminal stage of POAG were noted.Conclusion. In the terminal stage of POAG, the interstitial spaces of the retina are increased and the volume density of blood vessels is dropping. The increased volume density of luminal and basal caveolae and the formation of transendothelial channels in the cytoplasm of blood capillary endotheliocytes indicate the growth of transcytosis and the permeability of the blood-retinal barrier.

The aim: to conduct a clinical and diagnostic evaluation of the effectiveness of the administration of a complex of drugs in the region of hemolymphocirculation in optical neuropathy in patients with edematous form of endocrine ophthalmopathy.Materials and methods. The results of diagnosis and treatment of 31 patients (61 eyes) with optical neuropathy on the background of edematous exophthalmos in endocrine ophthalmopathy were analyzed. To identify hidden forms of optical neuropathy, such studies were prescribed as computer microperimetry on a confocal infrared ophthalmoscope, a complex of electroretinographic studies: registration of maximum ERG, oscillatory potentials. Latent forms of optical neuropathy were detected in 22 cases. In 9 cases, there were obvious forms of optical neuropathy. All patients underwent a 10-day course of intensive complex treatment, consisting of injections into the region of hemolymphocirculation (projection of the pterygoid fossa) No. 6–8 (No. 3–4 on each side) with an interval of 24 hours of a drug mixture, the formulation of which included Lidocaine 20 mg, Dexazone 4 mg, Hemase 3000 UNITS, Dalargin 1 mg.Results. In all patients, there was a pronounced positive dynamics in the form of a significant (from 0.6 to 1.0) increase in visual acuity, a decrease in exophthalmos from (2.0 to 3.0 mm), an increase in color and contrast sensitivity.Conclusions. Due to the violation of venous and lymphatic outflow due to thickening of extraocular muscles and retrobulbar fiber, injections into the hemolymphocirculation region (projection of the pterygoid fossa) of drugs with a wide range of decongestant and metabolic effects are justified, effective and safe.

Purpose. To show the effectiveness and safety of the correction of residual ametropia by the PRK method after previously performed keratorefractive operations LASIK, FemtoLASIK, ReLEx SMILE. Material and methods. All patients were divided into 3 groups: Group 1 – previously performed LASIK surgery with insufficient stroma thickness under the valve and patients with unknown parameters of the corneal flap operated in third-party clinics 35 patients (50 eyes), age 46.3 ± 21.9 years, the value of the spherical equivalent – 2.41 ± 1.45. Group 2 – patients after femtolasik 6 patients (6 eyes), age 30 ± 8.3 years, spherical equivalent value – 1.83 ± 0.76. Group 3 – patients after ReLExSMILE 8 patients (11 eyes), age 33 ± 4.4 years, spherical equivalent value – 0.88 ± 0.66. The minimum period after the initial keratorefractive surgery was 1 year, the maximum follow-up period was 4 years. Results. When analyzing the results 6 months after the operation of photorefractive keratectomy: group 1 – the value of the spherical equivalent – 0.26 ± 0.46; group 2 – 0.11 ± 0.2; group 3 – 0.19 ± 0.34. In Group 1, 4 patients (5 eyes) had a loss of lines (3 patients (4 eyes) – 1 line, 1 patient (1 eye) – 2 lines) of maximum visual acuity, all other patients had UDVA to CDVA before the operation of PRK. In groups 2 and 3, the maximum figures of visual acuity were taught. Conclusions. The use of transepithelial photorefractive keratectomy allows to obtain high clinical and functional results, is a safe, easy-to-perform, universal and highly effective method of correcting residual ametropias after various previously performed keratorefractive operations. Keywords: PRK, PRK after LASIK, PRK after FemtoLASIK, PRK after SMILE.

The medical and social significance of one of the most severe inflammatory diseases of the posterior segment of the eye, which includes acute retinal necrosis (ARN), is determined by the high degree of disability of patients. ARN occurs in somatically healthy people, but occurs more often among people suffering from immunosuppressive diseases or receiving immunosuppressive therapy. The diagnosis of ARN is made on the basis of clinical examination data according to the criteria developed in 1994 by the American Society of Uveitis Specialists. The prognosis of ARN is relatively unfavorable: the pathological process leads to practical blindness in 64 %, in 60 % there is a significant decrease in visual acuity. ARN is complicated by the development of retinal detachment in 65–75 %. Treatment of ARN includes intensive antiviral, anti-inflammatory, immunomodulatory therapy. Purpose: description of the observation of a patient with acute retinal necrosis over a long period of time. Material and methods. A long-term clinical observation of a patient with acute retinal necrosis for the period from 2008 to 2021 is presented. The patient underwent a standard ophthalmological examination, optical coherence tomography of the macular area, and ultrasound. Results. The development of ARN in the patient was observed after ARVI. Antiinflammatory treatment of ARN and surgical treatment of complications were carried out in the conditions of the academician S.N. Fyodorov Federal State Institution National Medical Research Center «Intersectoral Research and Technology Complex «Eye microsurgery» Ministry of Health of the Russian Federation», Novosibirsk Branch. The presented clinical case allows us to demonstrate the severe course of the pathological process and the development of serious complications. Timely conservative, laser and surgical treatment, active dynamic monitoring, despite the unfavorable prognosis for vision, made it possible to maintain residual vision in the patient. Conclusions. ARN is an important problem that requires further study and development of effective methods for early diagnosis, treatment and prevention of recurrence of this disease. Keywords: acute retinal necrosis; anti-inflammatory treatment

КЛИНИЧЕСКИЙ СЛУЧАЙ ПАЦИЕНТКИ С ОСТРЫМ РЕТИНАЛЬНЫМ НЕКРОЗОМАрбеньева Н.С. 1

We studied the effect of conditioned media from limbal epithelial stem cells, fibroblasts, and corneal keratocytes on the functional activity of human limbal mesenchymal stem cells. It was shown that the conditioned media from limbal epithelial stem cells reduced proliferative activity and inhibited migration of limbal mesenchymal stem cells. In the conditioned media of limbal epithelial stem cells, increased concentrations of VEGF and TNFα and reduced concentration of BDNF, vimentin, and fibronectin were found. The conditioned medium from corneal stromal cells did not affect functional activity of mesenchymal stem cells in the limbus. These data contribute to the understanding of the interaction of cells in the limbal niche and with corneal cells essential for the maintenance of the cellular homeostasis in the cornea.

Purpose. To study the content of cytokines and matrix metalloproteinases in the intraocular fluid in patients with advanced stage of primary open-angle glaucoma. Material and methods. 50 patients were examined with a verified diagnosis of the advanced stage of POAG. The control group consisted of 30 patients with a diagnosis of uncomplicated cataract. The concentration of the determined biologically active molecules was carried out by the method of flow fluorimetry on a two-beam laser analyzer Bio-Plex 200, Bio-Rad, USA. Results. A statistically significant increase in the concentrations of IL-6, 8, 17, TGF-1, 2, 3, matrix metalloproteinase-2 and tissue inhibitors of matrix metalloproteinases-1, 2, 3, 4 was found in the intraocular fluid of patients with POAG. Conclusion. The data obtained indicate the significance of the local inflammatory process, as well as disturbances in the structure of the extracellular matrix and its remodeling in the mechanisms of POAG development. Keywords: primary open-angle glaucoma, intraocular fluid, cytokines, matrix metalloproteinases, tissue inhibitors of matrix metalloproteinases.

Background: glaucoma is one of the leading causes of blindness worldwide. Diagnosis of glaucoma at an early stage is challenging. Therefore, genetic factors predisposing to the development of primary open-angle glaucoma (POAG) are investigated. One of these predictors is tumor necrosis factor α (TNFα), a multifunctional proinflammatory cytokine involved in glaucoma pathogenesis. Point mutations in the regulatory region of the TNFα gene are hypothesized to be associated with POAG risk. Aim: to analyze the polymorphism of three positions of TNFα gene promoters and their complexes in West Siberian Caucasians with POAG and healthy volunteers. Patients and Methods: the study enrolled 401 individuals, i.e., 99 patients with POAG stage 2 and 302 individuals without POAG (randomized control group). All participants signed an informed consent form. Single nucleotide TNFα gene promoter polymorphism (rs361525, rs1800629, rs1800630) was analyzed. Genotyping was performed by restriction analysis of gene amplification products. Results: the occurrence of minor genotype TNF-308*АА was significantly higher in POAG (odds ratio 11.41, p=0.0011). The occurrence of two other genotypes demonstrated no significant differences between groups. Three complex genotypes were positively associated with POAG (TNF-863*CC:TNF-308*AA, TNF-308*AA:TNF-238*GG и TNF-863*CC:TNF-308*AA:TNF-238*GG). We failed to identify any single nucleotide polymorphism or complexes negatively associated with POAG. Conclusion: minor genotype TNF-308*АА is an essential factor of POAG pathogenesis. Two other polymorphic gene variants were associated with POAG as a part of complex genotypes. These findings demonstrate that potential polymorphic associations should be considered in the case-control analysis. Keywords: primary open-angle glaucoma, polymorphism, TNFα gene, gene promoter, complex genotypes. For citation: Shevchenko A.V., Prokof’ev V.F., Konenkov V.I. et al. Association of TNF-α gene promoter polymorphism with primary open-angle glaucoma. Russian Journal of Clinical Ophthalmology. 2022;22(1):11–15 (in Russ.). DOI: 10.32364/2311-7729-2022-22-1-11-15.