RU2151580C1 - Method for activating cornea endothelium proliferation by applying glecomen solution - Google Patents

Abstract

FIELD: medicine. SUBSTANCE: method involves applying solution of the following composition: heparin 5-100 units, glycosamineglycanes 0.01-0.5 mg, glycosamineglycanes-copper complex 0.005-0.15 mg in 1 ml of balanced salt solution. EFFECT: reduced endothelium loss in postoperative time; reduced cornea edema, increased endothelium density in cases of keratopathy. 6 cl

Description

 The invention relates to medicine, and more particularly to ophthalmology.

It is known that the possibilities of mitotic division in the endothelium of the human cornea are extremely limited. Endothelial regeneration, as a rule, occurs as a result of hypertrophy and an increase in the area of endothelial cells and their migration. With a decrease in the density of endothelial cells as a result of an accidental or operating injury, or eye disease below a critical level (less than 1000 cells / mm ² ), endothelial-epithelial dystrophy of the cornea can develop.

 A known method of activating the proliferation of the corneal endothelium using a solution containing heparin, copper ions and a balanced saline solution in the following ratio of components: heparin 200 units, copper ions 0.002 mg in 1 ml of 0.15 M phosphate buffer at pH 7.4 (US Pat. RF N2077878 dated 06/30/93). Such a solution has a stimulating effect on the activity of the endothelium, however, when using such a solution in a small percentage of cases the occurrence of hyphema due to an unbalanced amount of heparin is observed, which reduces the effectiveness of the effect.

 The objective of the invention is to provide a safe and effective method of activating the proliferation of corneal endothelium to prevent a decrease in endothelial density during surgical interventions, as well as the restoration of damaged corneal endothelium in postoperative keratopathy and the development of secondary endothelial-epithelial dystrophy.

 The technical result of the invention is to reduce the loss of endothelium in the postoperative period, and with postoperative keratopathy and the initial stage of secondary endothelial-epithelial dystrophy, a decrease in corneal edema, an increase in endothelial density and the disappearance of a patient’s discomfort.

The technical result is achieved by the fact that in the method of activating the proliferation of the corneal endothelium, comprising introducing a solution of heparin in a balanced saline solution into the anterior segment of the eye, according to the invention, the corneal endothelial proliferation is activated using the Glekomen solution of the following composition:
Heparin - 5-100 units.

Glycosaminoglycans - 0.01-0.5 mg
Glycosaminoglycans-copper complex - 0.005-0.15 mg
in 1 ml of a balanced saline solution, while a solution with a pH of 7.2 - 7.6 is used as a balanced saline solution in the following ratio of components, wt.%:
Sodium Chloride - 0.85 - 0.95
Monosubstituted sodium phosphate - 0.0025 - 0.0040
Disubstituted sodium phosphate - 0.0115-0.0135
Purified Water - Else
One of the variants of the invention is that during cataract extraction with implantation of an intraocular lens (IOL) or during implantation of an IOL with a refractive purpose, an activating solution is used to fill the anterior chamber of the eye at the end of the operation, or irrigation is carried out during the entire operation with an activating solution in an amount of 50-200 ml

 To reduce the loss of density of endothelial cells after surgery, the activating solution is administered subconjunctively at 0.3 ml daily for 3 days.

In the postoperative period with keratopathy, or with the development of the initial stage of secondary endothelial-epithelial dystrophy, the activating solution is administered subconjunctively at 0.3 ml daily for 5 to 10 days, or by phonophoresis through the cornea in a continuous mode for 10 minutes at an intensity of 0.2 - 0.3 W / cm ² using 5 ml of solution daily, while 5-8 sessions are carried out.

 The activating solution may additionally contain viscoelastic in an amount of 90 to 99 wt.% And they fill the front chamber of the eye at the initial stage of the operation.

 Heparin and glycosaminoglycans are naturally occurring polysaccharides built from repeating disaccharide components that typically contain amino sugar and uronic acid. In addition to heparin, glycosaminoglycans include keratan sulfate, chondroitin sulfate, hyaluronic acid, heparan sulfate, and dermatan sulfate. All glycosaminoglycans (GAGs) are polyanions due to the presence of acid sulfate groups or carboxyl groups of uronic acids in their structure. Many of their functional properties are associated with this feature of GAG. GAGs are integral components of connective tissue. They actively participate in water-salt metabolism, are able to reduce the inflammatory and exudative response in injuries, and stimulate the repair of damaged corneal tissue, and especially endothelium and keratocytes.

 Heparin, in addition to the listed properties, has a specific effect - it activates the fibroblastic growth factor from the descemetic membrane of the cornea, which in turn causes endothelial proliferation.

 The GAG-copper complex, obtained under physiological conditions (osmolarity and pH equal to tissue fluid), contains copper ions, which increase cell mobility, a necessary component of regeneration processes.

 Boundary concentrations of the components of the activating solution were determined experimentally. The concentration of heparin was expressed in units. anticoagulant activity, as when heparin is isolated from different sources, one antiviral agent corresponds to one weight unit. Therefore, the same weight units of heparin may have different anticoagulant activity, including one that leads to increased bleeding. This can lead to complications when using an activating solution. A heparin concentration of 5 u / ml is the minimum necessary to activate fibroblastic growth factor, and at a concentration of heparin above 100 u / ml, increased bleeding is observed.

 At a GAG concentration below 0.01 mg / ml, the effect of enhancing proliferation of rabbit corneal endothelium in the dosed injury zone was not observed. At a GAG concentration above 0.5 mg / ml, a decrease in the proliferation of keratocytes of the corneal stroma was observed by electron microscopy.

 At a concentration of the GAG-copper complex from 0.005 to 0.15 mg / ml, accelerated recovery of the rabbit corneal endothelium in the zone of the dosed defect was observed. At a concentration of the GAG-copper complex above 0.15 mg / ml, inhibition of the endothelium was observed, and at a concentration of the GAG-copper complex below 0.005 mg / ml, acceleration of endothelial recovery was not observed.

 Viscoelastics are viscous solutions of various substances used in ophthalmic surgery to maintain the anterior chamber volume and mechanically protect the corneal endothelium from injury during surgery. They are introduced into the anterior chamber of the eye at the initial stage of the operation and are displayed at the final stage. Currently, solutions of methyl cellulose derivatives are used in surgical practice - Occucoat (Eye World, v.2, N3, p.62, 1997), Visiton (US Pat. RF N 2114587), sodium hyaluronate - Provisc, Healon (J. Cataract Refract Surg ., 1998, 24, 1130 - 1135), sodium chondroitin sulfate - Viscoat (J. Cataract Refract Surg., 1998, 24, 678 - 683).

A solution for stimulating proliferation of the corneal endothelium is prepared as follows. First, a GAG complex with copper ions (GAG-copper) is obtained. To do this, to a solution of GAG with a concentration of 0.5 to 2.0 mg / ml in 0.1 M Tris-HCl buffer pH 7.4 add copper sulfate in a five-fold excess by weight, mix and leave at 4 - 8 ^o C for a day. The GAG-copper complex is isolated using gel chromatography, freeing itself from low molecular weight impurities. The concentration of the resulting GAG-copper solution was measured spectrophotometrically at a wavelength of 535 nm with a dye of 1,9-dimethylmethylene blue (Connective Tissue Research., 1982, v.9, p. 247-248.). Then, the components of the activating solution are mixed in concentrations according to the claims.

 The method is as follows.

 During surgical interventions involving opening the anterior chamber of the eye, such as cataract extraction or IOL implantation, etc., the anterior chamber is filled with the Glekomen activating solution at the final stage of the operation so that it directly contacts the corneal endothelium in the first postoperative hours, gradually replaced by the patient's secreted chamber moisture. With similar surgical interventions with an activating solution, irrigation of the anterior chamber can be carried out throughout the entire period of the operation.

 When performing the same surgical interventions with the Glekomen activating solution, additionally containing viscoelastic, the anterior chamber is filled in place of the traditionally used viscoelastics at the initial stage of the operation. At the final stage of the operation, said activating solution is withdrawn from the anterior chamber.

 To reduce the loss of endothelial cells in the early postoperative period after cataract extraction, implantation of IOL, etc., a course of subconjunctival injections of Glekomen activating solution is carried out after the operation every day for 3 days at 0.3 ml.

In cases of postoperative keratopathy or the initial stage of endothelial-epithelial dystrophy of the cornea, the Glekomen activating solution is administered subconjunctively daily for 5-10 days, 0.3 ml each. In similar cases, the Glekomen activating solution can be administered by phonophoresis. To this end, 5 ml of the solution is placed in a prosthetic bath placed on the cornea, and phonophoresis is carried out continuously for 10 minutes at an intensity of 0.2-0.3 W / cm ² . Phonophoresis is carried out daily for 5-8 days.

 Examples.

Example 1. Patient M., 53 L., the diagnosis of immature complicated cataract of the right eye. The density of endothelial cells (PEC) 2500-2600 cells / mm ² . Phacoemulsification with IOL implantation was performed. At the end of the operation, 0.3 ml of the Glekomen activating solution of the following composition was introduced into the anterior chamber: heparin 100 units. GAG 0.01 mg; GAG-copper complex 0.005 mg in 1 ml of balanced saline. 1 month after surgery, PEC 2300-2500 cells / mm ² , after 6 months PEC 2300-2500 cells / mm ² . The loss of PEC amounted to 5.9%, while the average loss of PEC during a similar operation without the use of Glekomen is 10.6%.

Example 2. Patient G., 68 l., The diagnosis is immature age-related cataract of the left eye, PEC 2 700-2 900 cells / mm ² . Performed phacoemulsification with IOL implantation. During the operation, at the stages of capsulorhexis, phacoemulsification of the nucleus, aspiration-irrigation of the lens masses and leaching of viscoelastic, Glekomen in the amount of 120 ml of the following composition was used as an irrigation solution: heparin 5 units, GAG 0.01 mg, GAG-copper 0 complex , 01 mg in 1 ml of balanced saline. 1 month after surgery, PEC 2600 - 2700 cells / mm ² , after 6 months 2500-2700 cells / mm ² . The loss of PEC amounted to 7.1%, while the average loss of PEC during similar operations without the use of Glekomen is 13.7%.

Example 3. Patient J., 70 l., The diagnosis of mature age-related cataract of the left eye, PEK 2500-2700 cells / mm ² . Extracapsular cataract extraction with IOL implantation was performed. At the end of the operation, 0.3 ml of Glekomen was administered subconjunctively daily for 3 days of the following composition: heparin 5 units, GAG 0.2 mg, GAG-copper complex 0.15 mg in 1 ml of balanced saline. 1 month after surgery, PEC 2500-2600 cells / mm ² , after 6 months PEC 2400-2600 cells / mm ² . The loss of PEC amounted to 3.8%, while the average loss of PEC during similar operations without the use of Glekomen is 12.5%.

Example 4. Patient S., 72 L., the diagnosis of secondary endothelial-epithelial dystrophy of the cornea of 1 degree, PEK 700-800 cells / mm ² , the thickness of the cornea in the Central optical zone of 607 microns. A course of conservative treatment was carried out, including daily phonophoresis of the cornea for 5 days using an activating Glecomen solution of the following composition: heparin 5 units. GAG 0.5 mg, GAG-copper complex 0.15 mg in 1 ml of balanced saline. To conduct phonophoresis, 5 ml of Glekomen of the indicated composition were placed in a phoretic bath and phonophoresis was carried out continuously for 10 minutes at an intensity of 0.2-0.3 W / cm ² . 1 month after treatment with PEC 1000-1100 cells / mm ² , 3 months after treatment, 800-1000. A repeated course of phonophoresis was carried out, after which PEC increased to 900-1000 cells / mm ² and continued to remain stable for 1 year. The thickness of the cornea in the central optical zone of the cornea after treatment decreased to 534 microns.

Example 5. Patient P., 72 L., the diagnosis of mature brown cataract of the right eye, PEK 1900-2100 cells / mm ² . Extracapsular cataract extraction with IOL implantation was performed. As an activating solution, Glekomen, additionally containing viscoelastic, of the following composition was used: heparin 100 units, GAG 0.01 mg, GAG-copper complex 0.15 mg, viscoelastic 0.99 g in 1 ml of balanced saline. 1 month after surgery, PEC 1800-1900 cells / mm ² , after 3 months PEC 1800-1900 cells / mm ² . The loss of PEC was 7.5%, while the average loss of PEC during similar operations without the use of Glekomen is 11.6%.

Claims (4)

1. A method of activating the proliferation of corneal endothelium, comprising introducing a solution of heparin in a balanced saline solution into the anterior segment of the eye, characterized in that the activating solution further comprises glycosaminoglycans and a complex of glycosaminoglycans with copper ions in the following ratio of components in 1 ml of balanced saline:
Heparin - 5 - 100 units. Glycosaminoglycans - 0.01 - 0.5 mg
Complex glycosaminoglycans - copper - 0.005 - 0.15 mg
2. The method according to p. 1, characterized in that as a balanced salt solution using a solution with a pH of 7.2 to 7.6 in the following ratio of components, wt.%:
Sodium Chloride - 0.85 - 0.95
Monosubstituted sodium phosphate - 0.0025 - 0.0040
Disubstituted sodium phosphate - 0.0115 - 0.0135
Purified Water - Else
3. The method according to claim 1 - 2, characterized in that during cataract extraction with IOL implantation or during IOL implantation with refractive purpose with an activating solution, the anterior chamber of the eye is filled at the end of the operation, or irrigation is carried out with an activating solution throughout the operation in an amount of 50 - 200 ml.  4. The method according to claims 1 and 2, characterized in that to reduce the loss of endothelial cells after surgery, the activating solution is administered subconjunctively at 0.3 ml daily for 3 days. 5. The method according to claims 1 and 2, characterized in that in the postoperative period with keratopathy or with the development of the initial stage of secondary endothelial-epithelial dystrophy, the activating solution is administered subconjunctively at 0.3 ml daily for 5 to 10 days, or by phonophoresis through the cornea in a continuous mode for 10 min, at an intensity of 0.2 - 0.3 W / cm ² using 5 ml of solution daily, while 5 to 8 sessions are carried out.  6. The method according to claim 1, characterized in that the activating solution, additionally containing viscoelastic in an amount of 90 to 99 wt.%, Fill the front chamber at the initial stage of cataract extraction.