CN115040535B - Application of sulfated hyaluronic acid in preparing eye drops for preventing corneal fibrosis and corneal scar - Google Patents
Application of sulfated hyaluronic acid in preparing eye drops for preventing corneal fibrosis and corneal scar Download PDFInfo
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Abstract
The invention discloses an application of sulfated hyaluronic acid in preparing eye drops for preventing corneal fibrosis and corneal scar. The eye drops comprise sulfated hyaluronic acid, a pH regulator and an osmotic pressure regulator. The main component of the eye drop, namely sulfated hyaluronic acid, can form a uniform, complete and stable coating on the surface of a corneal stroma, and has long eye retention time and high bioavailability. The coating can effectively block the permeation of cell factors excessively secreted for promoting epithelial healing to a stroma layer, and prevent the stroma cell and corneal fibroblast from being differentiated, so that corneal fibrosis and corneal scar formation are prevented. The eye drops have simple preparation process, stable quality and definite curative effect, can play an excellent role in preventing common ophthalmic complications such as corneal fibrosis and corneal scar, and have good clinical application prospect.
Description
Technical Field
The invention belongs to the field of eye drops, and particularly relates to application of sulfated hyaluronic acid in preparation of eye drops for preventing corneal fibrosis and corneal scar.
Background
Corneal fibrosis and corneal scar are common complications after corneal injury and keratoplasty, and the main symptoms of corneal fibrosis and corneal scar are represented by that normal corneal tissues are replaced by fibrotic tissues (extracellular matrix with disordered structure), so that the corneal transparency and vision of a patient are further affected. At present, researches on pathogenesis of corneal fibrosis and scar show that the main cause of the pathogenesis is defect of partial corneal epithelial tissue caused by mechanical injury of cornea after corneal operation. In order to accelerate the repair of corneal epithelial tissue defect and maintain the integrity of the corneal structure, corneal epithelial cells around the defect secrete a large amount of cytokines (mainly transforming growth factor beta 1, platelet-derived growth factor and the like) to promote corneal re-epithelialization. These excessive secreted cytokines penetrate damaged corneal basement membrane and the proelastic layer to enter the stromal layer, and induce normal corneal stromal cells and corneal fibroblasts to differentiate and proliferate into myofibroblasts. The differentiated myofibroblasts secrete disordered and disordered extracellular matrix to the periphery, and finally, corneal fibrosis and corneal scar are caused. Worldwide, corneal fibrosis and corneal scarring have been the major causes of corneal blindness.
In clinic, the prevention of corneal fibrosis and corneal scar is mainly controlled by using mitomycin C or hormone eye drops (such as tobramycin dexamethasone eye drops and the like). Although the medicinal eye drops are widely applied, the medicinal eye drops have inherent limitations. The eye drops have high eye clearance rate and short retention time, which leads to low bioavailability of the eye drops. Secondly, mitomycin C is a cell division inhibitor that inhibits the development of corneal fibrosis, corneal scarring, primarily by preventing excessive proliferation of corneal cells by over-secreted cytokines. The drug may have side effects such as increased intraocular pressure when applied, resulting in limited application.
Disclosure of Invention
The invention aims to provide application of sulfated hyaluronic acid in preparing eye drops for preventing corneal fibrosis and corneal scar. The eye drop contains sulfated hyaluronic acid, and can be used for preventing corneal fibrosis and corneal scar.
The purpose of the invention is realized by the following technical scheme.
The application of sulfated hyaluronic acid in preparing eye drops for preventing corneal fibrosis and corneal scar comprises sulfated hyaluronic acid, a pH regulator and an osmotic pressure regulator.
Preferably, the concentration of the sulfated hyaluronic acid in the eye drops is 0.5 to 5 mg/ml. More preferably, the concentration of the sulfated hyaluronic acid in the eye drops is 1.0 to 2.0 mg/ml, and most preferably 1.0 mg/ml.
Preferably, the molecular weight of the sulfated hyaluronic acid is 1 to 1000 kilodaltons. More preferably, the molecular weight of the sulfated hyaluronic acid is 100 to 150 kilodaltons.
Preferably, the sulfated hyaluronic acid has a sulfation degree of 1.0 to 3.0. More preferably, the sulfated hyaluronic acid has a degree of sulfation of from 2.5 to 3.0, most preferably 2.6.
Preferably, the sulfated hyaluronic acid includes, but is not limited to, metal salts such as sulfated hyaluronic acid, sulfated hyaluronic acid sodium salt, sulfated hyaluronic acid potassium salt, and the like.
Preferably, the metal salt of sulfated hyaluronic acid is a sulfated hyaluronic acid sodium salt or a sulfated hyaluronic acid potassium salt.
Preferably, the tonicity adjusting agent includes, but is not limited to, saccharide isotonic agents such as 5% aqueous glucose solution, salt isotonic agents such as 0.9% sodium chloride solution, organic isotonic agents such as glycerin, and the like.
Preferably, the pH adjusting agent includes, but is not limited to, phosphate buffer, borate buffer, citrate buffer, acetate buffer, tartrate buffer, carbonate buffer, amino acid buffer, and the like.
Preferably, the eye drops can also be added with a solution system consisting of at least one of bacteriostatic agent, stabilizing agent, viscosity increaser, solubilizer and protein protective agent. Furthermore, drugs including but not limited to tobramycin, dexamethasone and the like can be added into the eye drop system to prepare the eye drop which has multiple functions of preventing corneal fibrosis and corneal scar, resisting inflammation, resisting infection and the like.
Preferably, the dosage form of the eye drops can be aqueous solution, suspension, emulsion and the like. More preferably, the ophthalmic solution is provided in the form of an aqueous solution.
Preferably, the preparation of the eye drops comprises the following steps:
dissolving sulfated hyaluronic acid in physiological saline as osmotic pressure regulator, adding pH regulator, stirring to dissolve, adjusting pH to 7.0-7.5, sterilizing, and packaging.
Preferably, the sterilization mode is one of filter membrane sterilization, irradiation sterilization and high-temperature and high-pressure sterilization.
The principle of the invention is as follows: after the eye drops are dripped on the ocular surface, the sulfated hyaluronic acid can quickly form a uniform, complete and stable coating on the damaged corneal stroma on the ocular surface. The coating has high bioactivity, can be combined with over-secreted cell factors (transforming growth factor beta 1, platelet-derived growth factor and the like), prevents a large amount of the over-secreted cell factors from entering a corneal stroma layer to induce downstream reaction, and realizes effective prevention of corneal fibrosis and corneal scar.
Compared with the prior art, the invention has the following beneficial effects:
(1) Compared with the existing medicine eye drops, the eye drop of the invention contains sulfated hyaluronic acid, can quickly form a uniform, complete and stable coating on the damaged corneal stroma, has long retention time on the ocular surface and high bioavailability.
(2) The eye drops of the invention have simple preparation process, strong repeatability and stability and definite curative effect.
(3) The eye drops can play an excellent role in preventing common complications such as corneal fibrosis and corneal scar, and have important scientific research significance and good clinical application prospect.
Drawings
FIG. 1 is a schematic diagram of the application of the eye drops for preventing corneal fibrosis and corneal scar.
FIG. 2 is the high resolution spectrum of sulfur element 2p orbit of rabbit corneal stroma X-ray photoelectron spectrum with sulfated hyaluronic acid coating obtained in example 5.
FIG. 3 is the high resolution spectrum of sulfur element 2p orbit of rabbit corneal stroma X-ray photoelectron spectrum with sulfated hyaluronic acid coating obtained in example 7.
FIG. 4 is the high resolution spectrum of sulfur element 2p orbital of X-ray photoelectron spectrum of rabbit corneal stroma with sulfated hyaluronic acid coating obtained in example 8.
FIG. 5 is a graph showing the results of CCK8 proliferation on type I collagen membrane with sulfated hyaluronic acid coating by corneal epithelial cells obtained in example 9.
FIG. 6 is a graph of the internal TGF-. Beta.1 content of the stroma of the rabbit corneas of different coatings obtained in example 10.
FIG. 7 is a graph of the TGF-. Beta.1 content within the stroma of different coated rabbits obtained in example 11.
FIG. 8 is a graph of the TGF-. Beta.1 content within the stroma of different coated rabbits obtained in example 12.
FIG. 9 is a graph showing the results of observation in the bright field of corneal slit lamp 28 days after the rabbit corneal fibrosis model efficacy test in example 13.
FIG. 10 shows the results of 28 days post-operative corneal optical coherence tomography observation of rabbit corneal fibrosis model in example 13.
FIG. 11 is a graph of immunohistochemical staining of corneal alpha-smooth actin 28 days after the rabbit corneal fibrosis model efficacy experiment of example 13.
Detailed Description
The technical solution of the present invention will be further described with reference to the following examples and drawings, but the embodiments of the present invention are not limited thereto.
The application principle diagram of the eye drops for preventing corneal fibrosis and corneal scar is shown in figure 1.
Example 1
50.0 mg of sulfated hyaluronic acid (molecular weight of 100-150 kilodalton, degree of sulfation of 2.6) is dissolved in 50 ml of physiological saline, and then 0.1 g of sodium dihydrogen phosphate is added and fully stirred for dissolution. Adjusting the pH to 7.0 to 7.5 by using a proper amount of sodium hydroxide, filtering, sterilizing and subpackaging to obtain the sulfated hyaluronic acid eye drops with the concentration of 1.0 mg/ml.
Example 2
250.0 mg of sulfated hyaluronic acid (molecular weight of 100-150 kilodalton, degree of sulfation of 2.6) is dissolved in 50 ml of physiological saline, and then 0.1 g of sodium dihydrogen phosphate is added and fully stirred for dissolution. Adjusting the pH to 7.0-7.5 by using a proper amount of sodium hydroxide, filtering, sterilizing and subpackaging to obtain the sulfated hyaluronic acid eye drops with the concentration of 5.0 mg/ml.
Example 3
25.0 mg of sulfated hyaluronic acid (molecular weight of 100 to 150 kilodaltons, degree of sulfation of 2.6) was dissolved in 50 ml of physiological saline, and then 0.1 g of sodium dihydrogen phosphate was added thereto and sufficiently stirred and dissolved. Adjusting the pH to 7.0 to 7.5 by using a proper amount of sodium hydroxide, filtering, sterilizing and subpackaging to obtain the sulfated hyaluronic acid eye drops with the concentration of 0.5 mg/ml.
Example 4
50.0 mg of sulfated hyaluronic acid (molecular weight of 100-150 kilodalton, degree of sulfation of 2.6) is dissolved in 50 ml of physiological saline, 10 mu g of benzalkonium chloride (bacteriostatic agent) and 0.1 g of sodium dihydrogen phosphate are added and fully stirred and dissolved. Adjusting the pH to 7.0-7.5 by using a proper amount of sodium hydroxide, filtering, sterilizing and subpackaging to obtain the sulfated hyaluronic acid eye drops with the concentration of 1.0 mg/ml and the concentration of 3.0 mg/ml tobramycin.
Example 5
Taking fresh rabbit corneal tissues, removing corneal epithelium layer, front elastic layer, back elastic layer and inner cortex layer, and simulating the exposed corneal stroma surface after corneal surgery or mechanical injury by using the residual corneal stroma layer.
The eye drops prepared in example 1 were dropped on the surface of rabbit corneal stroma for 30 seconds, rinsed with distilled water, and then air-dried. The surface layer of the obtained corneal stroma is analyzed for sulfur element change by X-ray photoelectron spectroscopy. FIG. 2 is a high resolution spectrum of sulfur element 2p orbit of rabbit cornea stroma X-ray photoelectron spectrum with sulfated hyaluronic acid coating. The results show that the sulfur 2p peak corresponding to the sulfate group appears in the high resolution spectrum of the sulfur 2p on the surface of the substrate with the coating, and the successful formation of the sulfated hyaluronic acid coating is indicated.
Example 6 visualization of sulfated hyaluronic acid coating formation Process
In order to realize the visualization of the sulphated hyaluronic acid coating, a trace amount of 6-aminofluorescein was grafted on the sulphated hyaluronic acid by: 100.0 mg of sulfated hyaluronic acid (molecular weight of 100-150 kilodaltons, degree of sulfation of 2.6) is dissolved in 50 ml of physiological saline, and 10.0 mg of 6-aminofluorescein (the solubility of 6-aminofluorescein in water is low, and a small amount of undissolved 6-aminofluorescein should be added in a solution system). After stirring sufficiently, 69.4 mg of 1-ethyl- (3-dimethylaminopropyl) carbodiimide was added to the solution, and the reaction was carried out for 24 hours. After the reaction is finished, insoluble substances are removed by centrifugation, and the residual liquid is dialyzed for 72 hours by distilled water in a dialysis bag with the molecular weight cutoff of 3000 and then is freeze-dried for standby. The whole processes of the grafting reaction and the dialysis are carried out in a dark place.
50.0 mg of the prepared 6-aminofluorescein grafted sulfated hyaluronic acid is dissolved in 50 ml of normal saline, and 0.1 g of sodium dihydrogen phosphate is added into the solution and fully stirred and dissolved. Adjusting the pH to 7.0-7.5 by using a proper amount of sodium hydroxide, filtering, sterilizing and packaging.
The obtained eye drop is dripped on the surface of rabbit corneal stroma, and the formation process of the sulfated hyaluronic acid coating on the surface of the corneal stroma is observed by a laser confocal microscope. The results show that sulfated hyaluronic acid can form a uniform, integral coating on the corneal stroma surface within 30 seconds of instillation. The sulfated hyaluronic acid coating formed by soaking for 30 minutes is consistent with the coating formed by soaking for 30 seconds, which shows that the coating is very stable under the eye washing condition, long in retention time and high in bioavailability.
Example 7
Taking fresh rabbit corneal tissues, removing corneal epithelium layer, front elastic layer, back elastic layer and inner cortex layer, and simulating the exposed corneal stroma surface after corneal surgery or mechanical injury by using the residual corneal stroma layer.
The eye drops prepared in example 2 were dropped on the surface of the corneal stroma of rabbit for 30 seconds, rinsed with distilled water, and then air-dried. The surface layer of the obtained corneal stroma was analyzed for sulfur change by X-ray photoelectron spectroscopy.
250.0 mg of 6-aminofluorescein grafted sulfated hyaluronic acid prepared in example 6 was dissolved in 50 ml of physiological saline, and 0.1 g of sodium dihydrogen phosphate was added to the solution and dissolved by stirring thoroughly. Adjusting the pH to 7.0 to 7.5 by using a proper amount of sodium hydroxide, filtering, sterilizing and packaging. The resulting eye drops with a concentration of 5.0 mg/ml were dropped on the corneal stroma surface of rabbits for 30 seconds, washed, and the formation of the sulfated hyaluronic acid coating on the corneal stroma surface was observed by a laser confocal microscope.
FIG. 3 is a high resolution spectrum of sulfur element 2p orbit of X-ray photoelectron spectrum of sulfated hyaluronic acid coated rabbit cornea stroma when the concentration of sulfated hyaluronic acid component in the eye drop is 5.0 mg/ml. The results show that the peak of the sulfur element 2p corresponding to the sulfate group appears in the high resolution spectrum of the sulfur element 2p on the surface of the substrate, which indicates the successful formation of the sulfated hyaluronic acid coating.
Meanwhile, the formation condition of the coating is observed by a laser confocal microscope, and when the concentration of the sulfated hyaluronic acid component in the eye drops is 5.0 mg/ml, the sulfated hyaluronic acid can form a uniform and complete coating on the surface of the corneal stroma within 30 seconds after being dripped on the surface of the corneal stroma.
Example 8
Taking fresh rabbit corneal tissue, removing corneal epithelium layer, front elastic layer, back elastic layer and inner cortex layer, and simulating the exposed corneal stroma surface after corneal surgery or mechanical injury by using the residual corneal stroma layer.
The eye drops prepared in example 3 were dropped on the corneal stroma surface of a rabbit for 30 seconds, rinsed with distilled water, and then air-dried. The surface layer of the obtained corneal stroma is analyzed for sulfur element change by X-ray photoelectron spectroscopy.
25.0 mg of 6-aminofluorescein-grafted sulfated hyaluronic acid prepared in example 6 was dissolved in 50 ml of physiological saline, and 0.1 g of sodium dihydrogen phosphate was added to the solution and dissolved therein with stirring. Adjusting the pH to 7.0 to 7.5 by using a proper amount of sodium hydroxide, filtering, sterilizing and packaging. The resulting eye drops at a concentration of 0.5 mg/ml were dropped on the corneal stroma surface of rabbits for 30 seconds, followed by rinsing and observation of the formation of the sulfated hyaluronic acid coating on the corneal stroma surface by a laser confocal microscope.
FIG. 4 is a high resolution spectrum of sulfur element 2p orbit of X-ray photoelectron spectroscopy of stroma of rabbit cornea coated with sulfated hyaluronic acid when the concentration of sulfated hyaluronic acid component in the eye drops is 0.5 mg/ml. The results show that the peak of the sulfur element 2p corresponding to the sulfate group appears in the high resolution spectrum of the sulfur element 2p on the surface of the substrate, which indicates the successful formation of the sulfated hyaluronic acid coating.
Meanwhile, the formation condition of the coating is observed by a laser confocal microscope, and when the concentration of the sulfated hyaluronic acid component in the eye drops is 0.5 mg/ml, the sulfated hyaluronic acid can form a uniform and complete coating on the corneal stroma within 30 seconds after being dripped on the surface of the corneal stroma.
Example 9 Biosafety of sulfated hyaluronic acid coatings
A sterile circular type I collagen membrane having a diameter of 1 cm was immersed in 10 ml of the eye drop solution prepared in example 1 for 5 minutes, and rinsed with PBS to obtain a type I collagen membrane having a sulfated hyaluronic acid coating. Corneal epithelial cells at 10000/cm 2 The inoculation density of (2) was inoculated thereon at 37 ℃ with 5% CO 2 Then, the culture was carried out. CCK8 experiments were carried out at 24 hours, 48 hours and 72 hours of culture, respectively. Figure 5 shows the result of proliferation of CCK8 by corneal epithelial cells on collagen type I membrane with sulfated hyaluronic acid coating. The result shows that the corneal epithelial cells can normally proliferate on the collagen membrane I with the sulfated hyaluronic acid coating, the proliferation speed is not obviously different from that of a control group, and the sulfated hyaluronic acid coating is proved to have good biocompatibility.
EXAMPLE 10 sulfated hyaluronic acid component concentration of 1.0 mg/ml in eye drops, the sulfated hyaluronic acid coating formed had a barrier effect against permeation of TGF-. Beta.1 (transforming growth factor. Beta.1)
The rabbit corneal stroma with a sulfated hyaluronic acid coating prepared in example 5 was immersed in 5 ml of PBS solution containing 30 ng/ml TGF-. Beta.1 for 1 hour. After completion of the immersion, residual TGF-. Beta.1 on the surface was washed away with PBS, and 30 μm corneal stroma on the surface was removed with a surgical instrument. The tissue thus obtained was homogenized and then the TGF-. Beta.1 content was determined. The same procedure was performed on uncoated rabbit corneal stroma as a control experiment. FIG. 6 is a graph showing the TGF-. Beta.1 content inside the stroma of rabbit corneas of different coatings. The results show that the content of TGF-beta 1 in the corneal stroma of the sulfated hyaluronic acid coating group is obviously reduced compared with that of the uncoated group, and the sulfated hyaluronic acid coating layer can effectively prevent TGF-beta 1 from penetrating into the corneal stroma.
EXAMPLE 11 sulfated hyaluronic acid component concentration of 5.0 mg/ml in eye drops, the sulfated hyaluronic acid coating formed had a barrier effect against permeation of TGF-. Beta.1 (transforming growth factor. Beta.1)
The rabbit corneal stroma with a sulfated hyaluronic acid coating prepared in example 7 was immersed in 5 ml of PBS solution containing 30 ng/ml TGF-. Beta.1 for 1 hour. After completion of the immersion, residual TGF-. Beta.1 on the surface was washed away with PBS, and 30 μm corneal stroma on the surface was removed with a surgical instrument. The obtained tissue is homogenized and then the content of TGF-beta 1 in the tissue is determined. The same procedure was performed on uncoated rabbit corneal stroma as a control experiment. FIG. 7 is a graph showing the TGF-. Beta.1 content inside the stroma of rabbit corneas of different coatings. The results show that the content of TGF-beta 1 in the corneal stroma of the sulfated hyaluronic acid coating group is obviously reduced compared with that of the uncoated group, and the sulfated hyaluronic acid coating layer can effectively prevent TGF-beta 1 from penetrating into the corneal stroma.
EXAMPLE 12 sulfated hyaluronic acid component concentration of 0.5 mg/ml in an eye drop solution, the sulfated hyaluronic acid coating formed had a barrier effect against permeation of TGF-. Beta.1 (transforming growth factor. Beta.1)
The rabbit corneal stroma with a sulfated hyaluronic acid coating prepared in example 8 was immersed in 5 ml of PBS solution containing 30 ng/ml TGF-. Beta.1 for 1 hour. After completion of the immersion, residual TGF-. Beta.1 on the surface was washed away with PBS, and 30 μm corneal stroma on the surface was removed with a surgical instrument. The tissue thus obtained was homogenized and then the TGF-. Beta.1 content was determined. The same procedure was performed on uncoated rabbit corneal stroma as a control experiment. FIG. 8 is a graph of the TGF-. Beta.1 content within the stroma of rabbit corneas from different coatings. The results show that the content of TGF-beta 1 in the corneal stroma of the sulfated hyaluronic acid coating group is obviously reduced compared with that of the uncoated group, and the sulfated hyaluronic acid coating layer can effectively prevent TGF-beta 1 from penetrating into the corneal stroma.
Example 13 rabbit corneal fibrosis model efficacy experiment
Establishing a rabbit cornea fibrosis model: after the New Zealand rabbit is anesthetized, the conjunctival sac is washed and eye anesthesia eye drops are dripped. The corneal tissue was cut with a slight rotation using a 5.0 mm corneal trephine positioned at the center of the cornea. Approximately 100 μm of corneal tissue was surgically removed and the thickness of the cut was determined by corneal a ultrasonography.
Experimental grouping: the experiment is divided into three groups, namely a control group, an experimental group and a comparison group.
The treatment method comprises the following steps: control group-dropwise adding tobramycin eye drops to prevent infection; experimental group-tobramycin was added dropwise to prevent infection, and the eye drops prepared in example 1 were added dropwise; control group-drop tobramycin dexamethasone eye drops to prevent infection and treat (clinical medication control). The eye drops are applied 4 times a day, and the interval between different eye drops is 10 minutes.
The slit lamp is used for photographing and observing the occurrence conditions of corneal fibrosis and corneal scar. As a result, as shown in fig. 9, the incidence of corneal fibrosis and scarring was lower in the experimental group using the sulfated hyaluronic acid ophthalmic solution than in the other groups 28 days after the administration. As can be seen from the results of optical coherence tomography (see FIG. 10), the highly reflective white bands of the experimental group are the slightest, indicating that the group of corneal neogenetic matrixes is more regular than the other two groups. Meanwhile, immunohistochemical staining was performed on the marker protein alpha-smooth actin for corneal fibrosis of 28-day-extracted cornea (see fig. 11), and the range of protein expression was greatly reduced in the experimental group compared with the other two groups. The observation results show that the eye drops can effectively prevent corneal fibrosis and corneal scar.
The invention is not the best known technology.
The above embodiments are only for illustrating the technical idea and features of the present invention, and the purpose of the present invention is to enable those skilled in the art to understand the content of the present invention and implement the present invention, and not to limit the protection scope of the present invention by this means. All equivalent changes and modifications made according to the spirit of the present invention should be covered within the protection scope of the present invention.
Claims (8)
1. The application of sulfated hyaluronic acid in preparing eye drops for preventing corneal fibrosis and corneal scar is characterized in that the eye drops comprise sulfated hyaluronic acid, a pH regulator and an osmotic pressure regulator;
after the eye drops are dripped on the ocular surface, the sulfated hyaluronic acid in the eye drops forms a uniform, complete and stable coating on the damaged corneal stroma on the ocular surface;
the molecular weight of the sulfated hyaluronic acid is 100 to 150 kilodaltons, and the sulfation degree is 2.6.
2. The use of the sulfated hyaluronic acid as claimed in claim 1, in the preparation of eye drops for preventing corneal fibrosis and corneal scar, wherein the concentration of the sulfated hyaluronic acid in the eye drops is 0.5 to 5.0 mg/ml.
3. The use of the sulfated hyaluronic acid as defined in claim 2, in the preparation of eye drops for preventing corneal fibrosis and corneal scar, wherein the concentration of the sulfated hyaluronic acid in the eye drops is 1.0 to 2.0 mg/ml.
4. Use of sulfated hyaluronic acid as defined in any of claims 1 to 3, in the preparation of eye drops for the prevention of corneal fibrosis and corneal scarring, wherein the sulfated hyaluronic acid comprises sulfated hyaluronic acid and metal salts thereof.
5. The use of sulfated hyaluronic acid as defined in claim 4, for the preparation of eye drops for the prevention of corneal fibrosis and corneal scarring, wherein the metal salt of sulfated hyaluronic acid is a sodium or potassium salt of sulfated hyaluronic acid.
6. Use of sulfated hyaluronic acid as defined in any of claims 1-3, in the preparation of ophthalmic solutions for the prevention of corneal fibrosis, corneal scarring, wherein the osmolality adjusting agent is at least one of an aqueous glucose solution, a sodium chloride solution and glycerol; the pH regulator is at least one of phosphate buffer solution, borate buffer solution, citrate buffer solution, acetate buffer solution, tartrate buffer solution, carbonate buffer solution and amino acid buffer solution.
7. The use of sulfated hyaluronic acid as claimed in any of claims 1 to 3, for the preparation of eye drops for the prevention of corneal fibrosis and corneal scarring, wherein a solution system consisting of at least one of a bacteriostatic agent, a stabilizer, a viscosity-increasing agent, a solubilizing agent and a protein protectant is further added to the eye drops; the dosage form of the eye drops is aqueous solution, suspension or emulsion.
8. Use of sulfated hyaluronic acid according to any of claims 1 to 3, for the preparation of ophthalmic solutions for the prevention of corneal fibrosis, corneal scarring, characterized in that the preparation of said ophthalmic solutions comprises the following steps:
dissolving sulfated hyaluronic acid in physiological saline as an osmotic pressure regulator, adding a pH regulator into the solution, fully stirring and dissolving, regulating the pH to 7.0 to 7.5, sterilizing and packaging.
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