RU2776218C1 - Method for treating chronic corneal erosions - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention relates to medicine, namely, to ophthalmology, and can be used for treating chronic corneal erosion. The method involves taking blood in sterile test tubes containing 3.8% sodium citrate solution, centrifuging the blood for 5 minutes at 1,500 rpm to produce autologous conditioned plasma or at 3,500 rpm to produce platelet-enriched plasma. 1 ml of the resulting plasma is applied to the corneal surface in layers by finely spraying over the entire surface of the cornea for 2 to 3 seconds, followed by drying the ocular surface with an air flow until protective plasma film dries and adheres to the cornea, so that at least 10 layers are applied according to the above scheme until a multilayer protective plasma film with a high the adhesion degree is created on the corneal surface.

EFFECT: possibility of reducing the time for regeneration of the corneal epithelium, preventing the recurrence of epithelial defects, corneal neovascularisation and turbidity due to the creation of a multilayer autologous protective film on the corneal surface and introduction of adhesive molecules, growth factors, dissolved fibrin, and cytokines.

1 cl, 3 dwg, 1 ex

Description

The invention relates to medicine, namely to ophthalmology, including experimental, and can be used to treat chronic corneal erosions both in experimental conditions on animals and in the clinic in the treatment of patients.

Clouding of the cornea due to injuries or inflammatory diseases of various etiologies in 33-40% of cases leads to visual disability. At the same time, monocular blindness after injuries, erosions and ulcers of the cornea, annually in the world is the outcome of 1.5-2 million people [Kulikov A.N., Chernysh V.F., Churashov S.V., Gavrilyuk I.O. On the use of an amniotic membrane to optimize corneal epithelialization in the early stages of a severe eye burn (clinical observation) // Modern technologies in ophthalmology. - 2017. - no. 3. - S. 130-132; Kulikov A.N., Churashov S.V., Gavrilyuk I.O., Tarabrina V.A., Platonov N.I., Muravieva O.A., Troyanovsky R.L. Experimental model of recurrent corneal erosion // Ophthalmology. - 2019. - Vol. 16. - No. 2. - S. 230-235.]. In this regard, the problem of visual rehabilitation of patients with traumatic, inflammatory and degenerative diseases of the cornea remains relevant.

Among inflammatory diseases of the anterior segment of the eye, chronic corneal erosions, including persistent corneal erosion (PER) and recurrent corneal erosion (CER), deserve special attention. Despite the advances in modern ophthalmology, chronic corneal erosions are difficult to treat and almost always lead to a significant decrease in visual acuity. The main feature of these erosions is the polymorphism of etiological factors and clinical course, which complicates timely diagnosis and prevents effective therapeutic measures aimed at stopping the inflammatory process and enhancing the processes of corneal regeneration [Kasparova E.A., Kasparov A.A., Amir M.R. . a. N., Marchenko N.R., Makarova M.A., Borodina N.R., Fedorov A.A., Smirennaja E.V. Recurrent corneal erosion // Recidivirujushhaja jerozija rogovicy. [Cataract & Refractive Surgery]. Kataraktal'naja i refrakcionnaja hirurgija. - 2010. - T. 1. - P. 51-53; Trufanov S.V., Malozhen S.A., Polunina E.G. Syndrome of recurrent corneal erosion (review) // Ophthalmology. - 2015. - T. 12. - No. 2. - From 4-12.].

At the same time, modern ideas about the pathogenesis of PER and ER indicate that, regardless of etiology, the main cause of the development of the disease is the inferiority of the basement membrane and the failure of the "adhesion complex" of the epithelium to the Bowman's membrane, therefore, the main goal of the treatment of PER is to stimulate repair and regeneration, as well as the restoration of a full-fledged "adhesion complex" of the basement membrane. [Tarabrina V.A. Experimental model of recurrent corneal erosion // Proceedings of the Russian Military Medical Academy. - 2019. - Vol. 2. - No. S1. - S. 205-208.].

The use of fetal epithelial cells and keratoblasts, amniotic membrane, and platelet-rich plasma (PRP) currently seem to be the most promising approaches to the treatment of patients with chronic corneal erosions.

ObTP is a concentrate containing up to 1000×109 cells/l, i.e. 3-4 times more than in undiluted blood. Human platelets, in turn, have a pronounced regenerative effect due to alpha granules, which are a natural depot of growth factors: platelet-derived growth factor (PDGF), transforming growth factor (TGF-b), insulin-like growth factor (IGF-I, IGF- II), fibroblast growth factor (FGFb), epidermal growth factor (EGF) and vascular endothelial growth factor (VEGF), affecting the restoration of damaged epithelium by enhancing the processes of proliferation, migration and differentiation of epithelial cells [Fedoseeva E.V., Chentsova E. .V., Borovkova N.V., Alekseeva I.B., Romanova I.Yu. Morphofunctional features of platelet-rich plasma and its application in ophthalmology // Ophthalmology. - 2018. - T. 15. - No. 4. - S. 388-393]. At the same time, ObTP contains not only growth factors, but also adhesive molecules (fibrin, fibronectin and vitronectin) and cytokines that stimulate repair and anabolic processes in damaged tissues, and also have an anti-inflammatory effect [Fernández-Barbero, Juan Galindo-Moreno, Pablo Avila-Ortiz, Gustavo Caba, Octavio Sánchez-Fernández, E Wang, Hom-Lay. Flow cytometric and morphological characterization of platelet□ rich plasma gel // Clinical oral implants research. -2006. - T. 17. - no. 6. - C. 687-693.].

Since GTP is derived from the patient's own blood, its use does not carry the risk of parenteral transmission of infections such as HIV or hepatitis. The use of ObTP does not cause hyperplastic processes, carcinogenesis or tumor growth, since growth factors affect receptors located in the cell membrane, and not in the cell nucleus. Growth factor second messengers initiate normal gene expression rather than the pathological expression that underlies tumor development. Growth factors are not mutagens and do not block the feedback mechanisms of tissue repair and regeneration processes.

Thus, due to the presence of growth factors, adhesive molecules, and dissolved fibrin, ObTP can be considered as a promising substrate for the treatment of patients with chronic corneal erosions. However, at present, ObTP is used for this purpose only in the form of installations on the ocular surface, which is mediated with a number of difficulties:

1. accumulation of ObTP in the conjunctival fornix and further formation of a plasma clot in these places;

2. high consumption of obTP;

3. the difficulty of achieving long-term adhesion of the plasma clot on the corneal surface;

Under such conditions, the development of new methods for the treatment of chronic corneal erosions is very relevant. At the same time, for the treatment of this pathology, the possibility of using an easier-to-obtain autologous conditioned plasma (Autologous Conditioned Plasma (ACP)) has not been described in the literature at all.

For the treatment of chronic corneal erosion, there is a method in the literature for the treatment of recurrent corneal erosion of various origins [RF Patent No. subsequent replacement of it every 6-7 days, installation of picloxidine hydrochloride 0.05%, administration of dexpanthenol 5% and Balarpan-N 0.01%, 2 drops 4 times a day of each drug, with an interval of 5-7 minutes. Additionally, from the first day of treatment, a solution consisting of 1 ml of Oftandexamethasone diluted in 10 ml of Oftolik is instilled in a decreasing pattern: 1st week - 4 times a day, 2nd week - 3 times a day, 3rd week - 2 times per day, 4th week - 1 time per day. The installation of MKL and the introduction of picloxidine hydrochloride 0.05%, Balarpan-N 0.01% and dexpanthenol 5% are carried out until the cornea is completely epithelized.

The disadvantages of this method are the duration and complexity of the treatment regimen, the need for an intensive regimen of installations and a large number of drugs, which increases the risk of developing individual intolerance to the drugs used.

A known method for the treatment of corneal erosion [RF Patent No. 2173122 "Method of treating corneal erosion", 2001], according to which a multilayer layer of allogeneic or xenogenic epithelial cells is grown on the inner surface of the contact lens. The lens is used, on the one hand, as a matrix and cell carrier, and on the other hand, as a method for fixing the graft.

The disadvantage of this method is the complexity and high cost of cell cultivation, which requires the use of complex expensive equipment, consumables and techniques, which minimizes the chances for the widespread introduction of this method of treatment.

Closest to the proposed invention is a method for the treatment of chronic corneal erosion of herpetic etiology [RF Patent No. 2653260 "Method of treating chronic corneal erosion of herpetic etiology", 2017], which is performed in two stages. The first stage includes antiviral, antibacterial, reparative therapy for 3 weeks. At the second stage of treatment, a course of instillations of platelet-rich plasma is carried out 6 times a day in combination with a subconjunctival injection of platelet-rich plasma once a week. The number of procedures is from 3 to 5 times.

The main disadvantages of this method are:

1. the impossibility of providing conditions for adhesion of autoplasma on the corneal surface due to rinsing or dilution with lacrimal fluid, which in some cases leads to the absence of a clinical effect;

2. the impossibility of creating a multilayer autologous film with a high degree of adhesion;

3. The accumulation of ObTP in the conjunctival fornix and the further formation of a plasma clot in these places reduces the effectiveness of the treatment due to the deformation of the plasma film on the corneal surface;

4. uneconomical use of obTP due to its high consumption;

5. difficulty in achieving long-term adhesion of the plasma clot on the corneal surface;

6. The method is positioned for the treatment of chronic corneal erosions of herpetic etiology.

The technical result when using the invention is to reduce the time of regeneration of the corneal epithelium, prevent recurrence of epithelial defects, neovascularization of the cornea, as well as its clouding due to the creation of a multilayer autologous protective film on the surface of the cornea and the introduction of adhesive molecules, growth factors, dissolved fibrin and cytokines.

This technical result is achieved by the fact that in a method for treating chronic corneal erosion, including blood sampling into sterile tubes containing 3.8% sodium citrate solution, its centrifugation for 5 minutes at 1500 rpm to obtain autologous conditioned plasma and at 3500 rpm / min to obtain platelet-rich plasma and use the resulting autologous plasma, 1 ml of the obtained autoplasma is applied to the corneal surface in layers by finely dispersed spraying it over the entire surface of the cornea for 2-3 seconds and further drying the ocular surface until the protective film dries and adheres to the cornea from autoplasma so that at least 10 layers are applied according to this scheme until a multilayer autologous protective film with a high degree of adhesion is reached on the corneal surface.

The proposed method can significantly reduce the time of regeneration of the corneal epithelium, prevent recurrence of epithelial defects, neovascularization and clouding of the cornea in the optical region. The therapeutic effect of the method is based on the more rapid formation of a normal "adhesion complex" between the corneal epithelium and Bowman's membrane. It has been experimentally shown that layer-by-layer application of autologous plasma alternating with drying of the ocular surface makes it possible to achieve complete epithelization of the cornea by the 7th day. This autoplasma spray exposure is sufficient to completely cover the cornea with a thin layer of protective film, while it has been experimentally proven that a 10-layer coating provides the claimed result.

The invention is illustrated by the following figures: FIG. 1 - method of dispersion of autologous plasma; in fig. 2 shows a photograph of a rabbit's eye on the 3rd day after irrigation of autologous plasma; in fig. 3 - the same, on the 7th day.

The implementation of the method for the treatment of chronic corneal erosions will be described further on the example of a pilot study. At the preparatory stage, blood was taken from an experimental animal (Chinchilla rabbit) into sterile test tubes containing 3.8% sodium citrate solution, after which the blood was centrifuged for 5 min (at 1500 rpm to obtain ACP and at 3500 rpm to obtain ObTP), and 1 ml of the obtained autoplasma was applied to the corneal surface in layers by finely dispersed spraying over the entire surface of the cornea for 2-3 seconds and further drying of the eye surface until the protective film from autoplasma dries and adheres to the cornea so that, according to this scheme, at least 10 layers were applied until a multilayer autologous protective film with a high degree of adhesion was reached on the corneal surface, while excess autoplasma from the conjunctival fornix was blown away by an air stream at the stage of drying the ocular surface, which prevented the formation of a plasma clot in these places, and its fine dispersion repeatedly reduced consumption, which in preparation ovitive stage allowed to take a smaller amount of autologous blood (Fig. one).

The essence of the invention is illustrated by the following example. The study was performed on 16 rabbits (32 eyes). At the first stage, a model of chronic corneal erosion was created for all animals according to our earlier patent for the invention [RF Patent No. 2709834 "Modeling method in the experiment of recurrent corneal erosion", 2019]. At the second stage, the animals were divided into four groups (4 rabbits each). In the 1st main group (MG), treatment was carried out according to the proposed method by spraying 1 ml of autologous plasma according to the following scheme: irrigation of the entire surface of the cornea with autologous plasma, after which the corneal surface was dried. According to this scheme, 10 layers of ObTP were applied. In the 2nd MG, autologous plasma was installed 1 drop every 1 min for 10 min (10 drops) once, in the 3rd MG, treatment was carried out according to the above method daily for 5 days. In the control group (CG), no autologous plasma treatment was used.

In the 1st OG, a significant acceleration of regeneration processes was observed. On the 3rd day of the experiment, a deep turbidity of the corneal stroma was noted in the form of a "cloud" due to its edema, and the deepithelialization area was 97.3% (Fig. 2) and had no significant differences from CG. Already on the 7th day of the experiment, complete epithelization of the cornea, the absence of newly formed vessels and opacities in the optical region were noted (Fig. 3).

In MG II, the absence of an epithelial defect was noted only from the 20th day of the experiment, while the growth of newly formed vessels over the entire surface of the cornea and the development of opaque opacity in the optical region were recorded up to the 30th day of the experiment.

In TG III, complete epithelialization of the cornea was noted on the 9th day of the experiment, which was accompanied by the absence of newly formed vessels and opacities in the optical region.

In the CG, the absence of an epithelial defect was noted only on the 34th day of the experiment, along with the persistence of turbidity and neovascularization up to the 86th day.

Thus, the proposed method can significantly reduce the time of regeneration of the corneal epithelium, prevent recurrence of epithelial defects, neovascularization and clouding of the cornea in the optical region by creating a multilayer autologous protective film on the corneal surface and introducing adhesive molecules, growth factors, dissolved fibrin and cytokines into the erosion area. .

Distinctive features that provide an advantage in the treatment of chronic corneal erosion according to the proposed method over existing analogues are as follows:

1. due to the alternation of spraying and drying of the eye surface, it is possible to provide conditions for adhesion of autoplasma on the surface of the cornea;

2. the method allows creating a multilayer autologous protective film with a high degree of adhesion on the corneal surface;

3. Excess autoplasma does not accumulate in the conjunctival fornix, but is blown away by air flow at the stage of drying the ocular surface, which prevents the formation of a plasma clot in these places;

4. Due to the fine spraying of autoplasma, its consumption is many times reduced, which makes it possible to take a smaller amount of autoblood at the preparatory stage;

5. the technical simplicity of the manipulation is combined with the efficiency of the formation of long-term adhesion of the plasma protective film on the corneal surface;

6. The method is intended for the treatment of chronic corneal erosions of various etiologies.

Claims (1)

A method for treating chronic corneal erosion, which includes taking blood into sterile tubes containing a 3.8% sodium citrate solution, centrifuging it for 5 minutes at 1500 rpm to obtain autologous conditioned plasma or at 3500 rpm to obtain platelet-rich plasma and the use of the resulting autologous conditioned or platelet-rich plasma, characterized in that 1 ml of the resulting plasma is applied to the corneal surface in layers by finely dispersed spraying it over the entire surface of the cornea for 2-3 seconds and further drying the eye surface with an air stream until it dries and adheres to the cornea protective plasma film so that at least 10 layers are applied according to this scheme until a multilayer protective plasma film with a high degree of adhesion is achieved on the corneal surface.

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