RU2780273C1 - Method for surgical treatment of keratectasia of various origins - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention relates to medicine, in particular to ophthalmology, and can be used for the surgical treatment of keratectasia of various origins. For this, 0.2 ml of a freshly isolated suspension of autologous mononuclear blood leukocytes is injected around the protrusion of the cornea, into the anterior third of the stroma of the cornea, at an angle of 30 degrees to the surface of the cornea, using a 25G needle. The cell suspension is injected into 4 sectors around the zone of keratectasia until a clearly defined infiltration of the anterior third of the stroma of the affected cornea appears. The procedure is performed once.

EFFECT: method provides stable clinical and functional results in the postoperative period and good patient tolerance of the developed treatment with minimal trauma to the ocular surface of the operated eye, which significantly reduces the risk of intra- and postoperative complications.

1 cl, 2 ex, 25 dwg

Description

The invention relates to medicine, in particular to ophthalmology, and can be used to treat primary and secondary keratectasia.

Keratectasia is a multifactorial disease of the cornea, which is based on the loss of structural integrity and thinning of the corneal stroma with the formation of its protrusion. The causes of the development of the disease may be previously performed keratorefractive and keratororeconstructive operations, corneal injuries. The appearance of corneal ectasia is accompanied by the development of symptoms of corneal syndrome, a significant decrease in visual acuity, and a significant deterioration in the quality of life of patients [1, 2].

Currently, there are various methods of treatment of this disease. So, for example, at the initial stage of keratectasia, correction with hard contact lenses and ultraviolet cross-linking of the cornea in various modifications are used, in advanced stages - penetrating and layered keratoplasty [3, 4]. However, given the polyetiology of the disease and the pathomorphological features of its course, the problem of treating keratectasia of various origins remains unresolved to date. Of particular interest, in our opinion, is cell therapy, the use of which stimulates reparative processes in the dystrophically altered cornea.

A known method of drug treatment of keratoconus, which consists in the epibulbar injection of a solution of leukinferon [5]. The drug is instilled into the conjunctival cavity of the diseased eye 1-2 drops 2 times a day at the same time for 10 days. According to clinical observations, against the background of the use of this method of treatment, apoptosis of keratocytes of the affected cornea is suppressed due to the antioxidant properties of leukinferon. However, it should be noted that this method of treating keratectasia still does not provide the proper depth of impact on the cornea, exerting a therapeutic effect mainly on the surface layers of the cornea. In addition, this method of treatment is effective only at the initial stage of the disease, providing a temporary result, followed by progressive protrusion of the cornea.

There is also known a method for the treatment of the initial stage of keratoconus [6], involving subconjunctival - near the limbus of the affected cornea - the introduction of dispersed biomaterial Alloplant. The drug is preliminarily diluted in physiological sodium chloride solution in a ratio of 30 mg of biomaterial per 0.5-1.5 ml of solution with the addition of 0.5 ml of dexamethasone solution, after which Alloplant subconjunctival injections are performed at 4-6 points around the limbus of the dystrophically changed cornea with an interval 1 time in 6-12 months. The disadvantages of this method are the need for repeated injections to maintain the therapeutic effect, as well as the allogeneity of the administered drug with the risk of developing allergic and autoimmune reactions.

Closest to the claimed technical solution in terms of technical essence and the achieved technical result is a method for treating initial keratoconus using the Bioplant material impregnated with Superlymph [7].

This method of treatment is carried out as follows. The material "Bioplast" impregnated with the immunotropic drug Superlymph is used - a complex of natural cytokines - interleukin - 1, 2, 6, tumor necrosis factor, etc., which has an activating effect on the cells of the limbal zone of the cornea. A strip of tissue 8.0 mm long and 3.0 mm wide is cut out of the Bioplast material, which is then soaked in a Superlymph solution for 15 minutes. Next, the prepared strip of Bioplast material is introduced into the sub-Tenon space of the affected eye through a 2.0 mm incision in the conjunctiva of the sclera at a distance of 3.0 mm from the limbus, near the zone of corneal ectasia.

The disadvantages of the presented method of treatment is the allogeneity of the Superlymph drug, the trauma of surgical intervention, a significant risk of intra- and postoperative complications (deployment of the biomaterial, the addition of a secondary infection). In addition, a strip of Bioplast material located 3.0 mm from the limbus cannot fully provide a sufficiently effective effect on the affected cornea due to its distance from it. It should also be noted that this method of treatment, according to the recommendations of the authors, is indicated for the treatment of only the initial stage of keratoconus.

The present invention is based on the task of creating a new method for the surgical treatment of keratectasia of various origins using cellular technologies, the use of which will ensure high and stable clinical and functional efficiency at various stages of the disease.

To solve this problem, a new method for the surgical treatment of keratectasia of various origins has been developed, including a single injection of a freshly isolated suspension of autologous mononuclear blood leukocytes into the stroma of the affected cornea.

The method is carried out as follows. Autologous mononuclear cells from the patient's blood are isolated immediately before surgery by density gradient fractionation on a ficoll-verografin separating solution. Isolation of autologous mononuclear cells is carried out as follows: beforehand, in the conditions of the treatment room, under the conditions of asepsis and antiseptics, blood is taken from the patient's cubital vein in the amount of 4.0-5.0 ml. The blood is placed in a sterile tube containing 1.0 ml of heparin solution. Heparinized blood is diluted 2.0 times with isotonic sodium chloride solution. The resulting suspension is layered on 3.0 ml of a mixture of ficoll-verografin (12 parts of 9% ficoll and 5 parts of 33.9% verografin with a density of 1.067-1.077 g/ml). The ratio of the volumes of the gradient and the suspension to be separated is 1:3 - 1:4.

The tube is centrifuged at room temperature for 15 min at 800g (2000 rpm). After centrifugation, the interphase layer containing mononuclear leukocytes and located between the plasma and the gradient is taken with a Pasteur pipette. Add 1.0 ml of isotonic sodium chloride solution, and the resulting suspension is again centrifuged for 7 min at 400g (1500 rpm) to wash out the cell suspension.

The purity of mononuclear cells obtained on a ficoll-verografin gradient is 96-98%. The viability of the cellular material is evaluated in the trypan blue test in the following way: a 0.1% eosin solution is preliminarily prepared on the basis of Ringer's solution. Next, a 0.1% solution of trypan blue is prepared on the basis of distilled water. After that, 1-2 drops of a fresh mixture of solutions of previously prepared dyes, taken in equal volumes, are added to a drop of the cell suspension. The resulting mixture is placed in the Goryaev chamber. When counting cells, the percentage of stained (dead) elements should be 1.5-2%, which would not exceed the allowable (no more than 3%) amount. The viability of the cell material in the trypan blue test was 97-98%.

In the conditions of the operating room after processing the surgical field and anesthesia of the patient's diseased eye with the help of instillations into the conjunctival cavity of 1-2 drops of 0.4% solution of oxybupracaine, retrobulbar anesthesia 2.0 ml of 2% solution of lidocaine hydrochloride around the zone of corneal ectasia at an angle of 30 degrees to the surface the cornea at the level of the anterior third of its stroma, a 25G needle is inserted, put on a 1.0 ml syringe with a freshly isolated suspension of autologous mononuclear blood leukocytes. Further, by introducing 0.2 ml of cell suspension in 4 sectors around the zone of keratectasia, a clearly defined infiltration of the anterior third of the stroma of the affected cornea is achieved. Then, under the conjunctiva of the sclera of the diseased eye of the patient, a solution of a broad-spectrum antibiotic in a standard dilution is injected and a monocular bandage is applied for 1 day. In the postoperative period, eye drops of a broad-spectrum antibiotic are instilled into the conjunctival cavity of the diseased eye according to the standard scheme.

The essence of the proposed technical solution is illustrated by drawings. Below is a brief description of the attached figures used to describe the implementation of the invention.

Figure 1 presents a biomicroscopic picture of the cornea of the diseased eye of a patient with secondary keratectasia at admission;

figure 2 shows an optical section of the cornea of the diseased eye of a patient with secondary keratectasia at admission;

figure 3 shows the OCT-tomogram of the cornea of the diseased eye of the patient with secondary keratectasia at admission;

figure 4 presents a biomicroscopic picture of the cornea of the diseased eye of a patient with secondary keratektasia on the 3rd day after surgery;

figure 5 shows the OCT-tomogram of the cornea of the diseased eye of the patient with secondary keratectasia on the 3rd day after surgery;

figure 6 presents a biomicroscopic picture of the cornea of the diseased eye of a patient with secondary keratectasia on the 10th day after surgery;

figure 7 shows an optical section of the cornea of the diseased eye of a patient with secondary keratectasia on the 10th day after surgery;

figure 8 shows the OCT-tomogram of the cornea of the diseased eye of the patient with secondary keratectasia on the 10th day after surgery;

figure 9 presents a biomicroscopic picture of the cornea of the diseased eye of a patient with secondary keratectasia 1 month after surgery;

figure 10 shows the OCT-tomogram of the cornea of the diseased eye of the patient with secondary keratectasia 1 month after surgery;

figure 11 presents a biomicroscopic picture of the cornea of the diseased eye of a patient with initial keratoconus upon admission;

Fig.12 shows the appearance of the cornea of the diseased eye of a patient with initial keratoconus when looking down at admission;

Fig.13 presents an OCT-tomogram of the cornea of the diseased eye of a patient with initial keratoconus upon admission;

Fig.14 presents a biomicroscopic picture of the cornea of the diseased eye of a patient with initial keratoconus on the 3rd day after surgery;

Fig.15 shows the appearance of the cornea of the diseased eye of a patient with initial keratoconus when viewed from top to bottom on the 3rd day after surgery;

Fig.16 presents an OCT-tomogram of the cornea of the diseased eye of a patient with initial keratoconus on the 3rd day after surgery;

Fig.17 presents a biomicroscopic picture of the cornea of the diseased eye of a patient with initial keratoconus on the 10th day after surgery;

Fig.18 shows the appearance of the cornea of the diseased eye of a patient with initial keratoconus when viewed from top to bottom on the 10th day after surgery;

Fig.19 presents an OCT-tomogram of the cornea of the diseased eye of a patient with initial keratoconus on the 10th day after surgery;

Fig.20 presents a biomicroscopic picture of the cornea of the diseased eye of a patient with initial keratoconus 1 month after surgery;

Fig.21 shows the appearance of the cornea of the diseased eye of a patient with initial keratoconus when viewed from top to bottom 1 month after surgery;

Fig.22 presents an OCT-tomogram of the cornea of the diseased eye of a patient with initial keratoconus 1 month after surgery;

Fig.23 presents a biomicroscopic picture of the cornea of the diseased eye of a patient with initial keratoconus 3 months after surgery;

Fig.24 shows the appearance of the cornea of the diseased eye of a patient with initial keratoconus when looking down 3 months after surgery;

Fig.25 presents an OCT-tomogram of the cornea of the diseased eye of a patient with initial keratoconus 3 months after surgery.

The advantages of the developed new method of surgical treatment of keratectasia of various origins are:

- the use of a suspension of cells of autologous origin from the patient's blood, which eliminates the risk of developing an autoimmune reaction, as well as the transmission of transmissible infections;

- the use of a freshly isolated cell suspension does not require its conservation and the provision of additional special storage conditions (temperature, humidity, oxygenation level, sterility), which significantly reduces the financial and economic costs of a medical institution;

- a single injection of a freshly isolated suspension of autologous mononuclear blood leukocytes into the stroma of the affected cornea is recommended during the treatment of keratectasia of various origins;

- the method of introducing a cell suspension into the anterior third of the affected cornea is technically simple in comparison with the presented prototypes and does not require special microsurgical skills;

- the introduction of a suspension of autologous mononuclear blood leukocytes into the anterior third of the cornea of the diseased eye provides a high concentration of biologically active substances in the immediate vicinity of the keratectasia zone, providing faster stabilization of the pathological process;

- the use of the proposed method of surgical treatment of keratectasia of various origins is possible at various stages of the disease, including advanced.

Case Study #1

Patient L., 64 years old. She complained of pain, photophobia, a significant decrease in vision in the left eye. These complaints were disturbing for 2 weeks, in connection with which the patient turned to an ophthalmologist, treatment was prescribed in the form of instillations of antibacterial, anti-inflammatory and reparative agents into the conjunctival cavity of the diseased eye. A few days later, the condition of the eye worsened, in connection with which the patient was recommended for emergency hospitalization in an ophthalmological hospital. During the collection of the patient's life history, it was found that about 30 years ago she underwent radial keratotomy for high myopia in both eyes.

Diagnosis upon admission to the ophthalmological clinic: secondary keratectasia of the left eye. Operated high myopia of both eyes.

On admission:

Vis OD = 0.7 does not correct (n/c)

Vis OS= 0.005 n/c

Level of intraocular pressure (IOP)

OD=18 mmHg, OS=22 mmHg

Biomicroscopy: OD - the eye is calm, on the periphery of the cornea in the superficial layers of the stroma, whitish, linear, radially located scars after keratotomy are found. The anterior chamber is of medium depth, the moisture is transparent, the reaction of the pupil to light is preserved, the initial opacities in the anterior and posterior cortex were determined in the lens. From the side of the fundus of the eye, no pathology was detected.

OS - moderate photophobia and lacrimation, blepharospasm, mixed injection of the eyeball. The optical section of the cornea is uneven in thickness, in the lower half of the cornea a pronounced edema of the epithelium and a conical protrusion of the stroma with its significant thinning at the apex are visualized, uneven edema and infiltration of the main substance, folds of the Descemet's membrane are detected perifocally. Radially on the periphery of the cornea, in the superficial layers of the stroma - whitish, linear scars after keratotomy (figure 1, figure 2). The anterior chamber is of medium depth, the moisture is transparent, the pupil is of medium width, the reaction to light is "live", in the lens there are initial opacities in the anterior and posterior cortex. From the side of the fundus of the eye, no pathology was detected.

According to optical coherence tomography (OCT) of the anterior segment of the left eye, the thickness of the cornea in the area of ectasia was 595 μm, around the zone of protrusion of the cornea - 894 μm, clusters of cystic cavities were found in the stroma of the cornea (figure 3).

The patient underwent surgical treatment of secondary keratectasia of the left eye by a developed method, including intrastromal injection of a suspension of autologous mononuclear blood leukocytes into the anterior third of the stroma of a dystrophically altered cornea. The operation was performed according to the presented technique, without complications.

In the postoperative period, broad-spectrum antibiotic eye drops were instilled into the conjunctival cavity of the operated eye according to the standard scheme.

During the observation of the patient in the postoperative period, the following dynamics of the clinical picture of the disease was revealed.

On the 3rd day after surgery:

Vis OS=0.1 n/a, palpation OS IOP level is normal.

Biomicroscopy of the left eye showed slight blepharospasm and mixed injection of the bulbar conjunctiva. The optical section of the cornea is uneven due to ectasia in the lower half, the edema of the anterior epithelium has decreased somewhat, the edema and infiltration of the stroma around the ectasia zone, the folds of the Descemet's membrane (figure 4) persisted. According to OCT of the cornea of the left eye, the thickness of the cornea in the ectasia zone decreased to 423 microns, around the ectasia zone - up to 673 microns due to a significant decrease in the number of cystic cavities in the thickness of the main substance, reducing them in area and volume (figure 5). From the side of the deeper parts of the operated left eye - no negative dynamics.

On the 10th day after the operation, the patient noted the absence of photophobia in the left eye and only slight pain, as well as some improvement in visual acuity of the operated eye.

Vis OS=0.2 n/c, IOP level OS=21 mm Hg. Art.

When biomicroscopy of the left eye - the anterior epithelium of the cornea is calm, the zone of ectasia in the lower third of the cornea significantly decreased in area and degree of protrusion, slight swelling and infiltration of the stroma, thin folds of the Descemet's membrane (Fig.6, Fig.7) remained around the pathological zone of the cornea. According to OCT of the cornea of the left eye, the thickness of the cornea in the ectasia zone was 414 µm, around it - 653 µm with single slit-like cystic cavities in the stroma of the membrane (Fig.8).

1 month after the surgical treatment of secondary keratectasia of the patient's left eye with the new proposed method, the patient noted the absence of complaints from the operated eye.

Vis OS=0.5 n/c, IOP level OS=20 mm Hg.

When biomicroscopy of the left eye - there was a slight unevenness of the optical section of the cornea due to residual ectasia in the lower third of the cornea, the anterior epithelium and stroma of the cornea are calm, single thin folds of the Descemet's membrane were determined (Fig.9). According to OCT data of the cornea of the left eye, the thickness of the cornea in the ectasia zone was 394 μm, perifocally - 633 μm, while the corneal stroma in the ectasia zone and around it was homogeneous, without signs of edema and cystic cavities (Fig. 10).

After 3, 6 months and 1 year after the surgical treatment of secondary keratectasia of the patient's left eye with the new proposed method, the clinical and functional results of the treatment corresponded to those achieved 1 month after the operation.

There were no complications from the operated eye during the entire follow-up period (1 year).

Case Study #2

Patient M., 31 years old. He complained of pain, photophobia, a sharp decrease in vision in the left eye. These complaints appeared about 2 days ago against the background of relative health. The patient went to the ophthalmological emergency room and, after examination by the on-duty ophthalmologist, was urgently admitted to the ophthalmological clinic.

Diagnosis at admission: acute keratoconus of the left eye, initial keratoconus of the right eye.

On admission:

Vis OD= 0.3 with sph +1.5 cyl -2.5 Ax 54=0.8

Vis OS= 0.05 n/c

IOP level OD=17 mm Hg, OS - palpation norm.

Biomicroscopy: OD - the eye is calm, the cornea is transparent, the anterior chamber is of medium depth, the moisture is transparent, the pupil reaction is "live", the lens is transparent. Examination of the fundus revealed no pathology.

OS - moderate photophobia and lacrimation, blepharospasm, mixed injection of the conjunctiva of the eyeball. The optical section of the cornea is uneven in thickness, the ectasia zone captures up to 2/3 of the cornea, edema and infiltration of the ground substance, folds of the Descemet's membrane are noted perifocally (Fig.11, Fig.12). The anterior chamber is of medium depth, the moisture is transparent, the reaction of the pupil to light is preserved, the lens is transparent. Examination of the fundus is difficult due to changes in the cornea, the reflex from the fundus is pink.

According to the B-scan data of the left eye, pathology of the fundus was revealed. According to OCT data of the cornea of the left eye, the average thickness of the cornea in the ectasia zone was 1594 μm, edema and cystic cavities were found in the ground substance (Fig. 13).

The patient underwent surgical treatment of acute keratoconus of the left eye with a new developed method. The operation was performed according to the presented technique, no complications were observed during the surgical intervention.

In the postoperative period, broad-spectrum antibiotic eye drops were instilled into the conjunctival cavity of the operated eye according to the standard scheme.

During the observation of the patient in the postoperative period, the following dynamics of the clinical picture of the disease was revealed.

On the 3rd day after surgical treatment:

Vis OS=0.05 n/a, IOP OS level - palpation is normal.

Biomicroscopy of the left eye showed slight blepharospasm and mixed injection of the bulbar conjunctiva. The optical section of the cornea is uneven due to ectasia in the lower half of the membrane, in the ectasia zone, in the thickness of the ground substance, a large cavity was determined. Perifocally, around the zone of ectasia, edema and infiltration of the stroma, folds of the Descemet's membrane were found (Fig.14, Fig.15). According to OCT data of the cornea of the operated eye, the thickness of the membrane in the ectasia zone was 1790 µm (Fig. 16). From the other parts of the operated eye - no negative dynamics.

On the 10th day after the intrastromal administration of a suspension of autologous mononuclear leukocytes, patient M. noted a significant decrease in pain in the left eye and photophobia, as well as some improvement in visual acuity of the diseased eye.

Vis OS= 0.08 n/c. IOP level OS =18 mm Hg.

Biomicroscopy of the left eye shows that the anterior epithelium of the cornea is calm, the ectasia zone in the lower third of the cornea has significantly decreased in terms of the degree of protrusion, the cavity in the thickness of the shell stroma has decreased in height. Around the ectasia, slight edema and infiltration of the main substance, thin folds of the Descemet's membrane remained (Fig. 17, Fig. 18). According to the pachymetry of the cornea of the left eye, the thickness of the cornea was 1428 μm. According to OCT of the anterior segment of the left eye, in the stroma of the main substance of the cornea, single cystic cavities were determined, which significantly decreased in size when compared with the initial data (Fig. 19).

One month after the surgical treatment of acute keratoconus with a new developed method, the patient noted the absence of complaints from the operated eye and improvement in the visual acuity and quality of the left eye.

Vis OS=0.1 n/c. IOP level OS = 21 mm Hg.

When biomicroscopy of the left eye - a slight unevenness of the optical section of the cornea due to residual ectasia in the lower third of the cornea. The anterior epithelium and stroma of the cornea are calm, single thin folds of the Descemet's membrane were determined (Fig.20, Fig.21). The index of pachymetry of the cornea of the operated eye was 821 microns. According to the OCT of the cornea of the left eye, the stroma of the cornea was homogeneous, without signs of edema and cystic cavities (Fig.22).

3 months after the surgical treatment of the patient with the new proposed method, there were no complaints.

Vis OS = 0.1 with sph -1.5 cyl -4.0 Ax 111=0.2-0.3

IOP level OS = 19 mm Hg.

Biomicroscopy of the left eye revealed a moderately intense clouding of irregular shape with clear boundaries in the lower half of the cornea, the surrounding corneal tissue was calm. Corneal ectasia was not observed (Fig.23, Fig.24). When performing OCT of the cornea of the operated eye, the pachymetry index was 718 μm, while the stroma of the cornea was homogeneous, without signs of edema and cystic cavities (Fig.25). From the side of the fundus - without features.

6 and 12 months after the surgical treatment of acute keratoconus in a patient with a new developed method, the clinical and functional results remained at the level achieved 3 months after treatment.

During the entire follow-up period (1 year), no complications were detected in the patient.

According to the generally accepted opinion [8], the action of cell therapy in the treatment of inflammatory and degenerative diseases of the cornea is based primarily on the trophic effect due to the secretory potential of the injected cells. For example, blood mononuclear leukocytes, according to scientists [8, 9], have the ability to synthesize and secrete a number of anti-inflammatory cytokines - transforming growth factor β-1, interleukin-1, -10, etc. Due to the action of these biologically active substances in the area of the pathologically altered cornea, the proliferation of cells of various populations, primarily of epithelial nature, is stimulated, as well as increased collagen production.

Another mechanism of action of cell therapy in the treatment of inflammatory and degenerative diseases of the cornea is the substitution effect - the injected cells actually fill the existing tissue defects in the areas of the damaged cornea.

The result of the combination of these effects of cell therapy in the treatment of inflammatory and degenerative diseases of the cornea is the stimulation of reparative regeneration of damaged corneal structures with a gradual improvement in the shape and curvature of the pathologically altered cornea. This, in turn, helps to eliminate the corneal syndrome and improve visual functions in patients of this category.

The technical result is a new method for the surgical treatment of keratectasia of various origins using cellular technologies, which has a higher clinical and functional efficiency.

EFFECT: invention makes it possible to exclude the use of allogeneic, as well as donor material in the surgical treatment of keratectasia of various origins. The presented method of surgical treatment is characterized by minimal trauma to the ocular surface of the operated eye, which can significantly reduce the risks of intra- and postoperative complications. The new method of surgical treatment is technically simple and does not require special microsurgical skills.

The use of the developed method for the surgical treatment of keratectasia provides stable clinical and functional results in the postoperative period, is well tolerated by patients and does not cause complications.

Sources of information

1. Balashevich L.I., Kachanov A.B., Golovatenko S.P. Ophthalmosurgery. 2009; 6:4-8,

2. Said A., Hamade I.H., Tabbara K.F. Saudi J. Ophthalmol. 2011; 25:225–230

3. Solodkova E.G., Remesnikov I.A. Modern approaches in the treatment of progressive keratectasia // Practical medicine. - 2012. - V. 4, No. 59. - S. 75-79.

4. Iomdina E.N. Biomechanical aspects of keratorefractive surgery and corneal crosslinking // Russian Pediatric Ophthalmology. - 2015-№ 4. - S. 32-37.

5. Patent RU2171659 "Method of drug treatment of keratoconus", IPC A61F 9/00, publ. 08/10/2001

6. Patent RU2421195, "Method of treating initial keratoconus", IPC А61F 9/00, A 61K 35/32, A61P 27/02, publ. 06/20/2011.

7. Patent RU2325141, "Method of treatment of initial keratoconus", IPC A61F 9/007, publ.27.05.2008.

8. Avetisov S.E., Subbot A.M., Antokhin A.I., Kasparova E.A., Kasparov A.A., Pavlyuk A.S. Personalized cell therapy in ophthalmology (II): cytokine profile of an autogenous cellular product. Cell transplantology and tissue engineering. - 2012. - V. 7, No. 1. S. 49-53.

9. Kado M, Tanaka R, Arita K, Okada K, Ito-Hirano R, Fujimura S, Mizuno H. Human peripheral blood mononuclear cells enriched in endothelial progenitor cells via quality and quantity controlled culture accelerate vascularization and wound healing in a porcine wound model. Cell Transplant.2018 Jul;27(7):1068-1079. doi: 10.1177/0963689718780307. Epub 2018 Jul 5. PMID: 29974793; PMCID: PMC6158547.

Claims (1)

A method for the surgical treatment of keratectasia of various origins, which consists in the local injection of a freshly isolated suspension of autologous mononuclear blood leukocytes, characterized in that the cell suspension of 0.2 ml is injected once into the anterior third of the corneal stroma around the zone of its protrusion using a 25G needle at an angle of 30 degrees to the surface of the cornea in 4 sectors around the zone of keratectasia until a clearly defined infiltration of the anterior third of the stroma of the affected cornea appears.

Images