KR20140019998A - Method for manufacturing hyaluronic acid derivatives sponge and hyaluronic acid derivatives manufactured thereby - Google Patents

Abstract

The present invention relates to a method for manufacturing hyaluronic acid derivative sponge and a hyaluronic acid derivative sponge manufactured thereby and more specifically, to a method for manufacturing hyaluronic acid derivative sponge which easily manufactures crosslinked hyaluronic acid derivative into a sponge form having excellent characteristic using a freeze drying method in order to have enhanced mechanical strength, no toxicity, and enhanced biodegradation control property and a hyaluronic acid derivative sponge manufactured thereby. The hyaluronic acid derivative sponge of the present invention has excellent biocompatibility, biodegradation control properties, and mechanical strength so that the hyaluronic acid derivative sponge can be used for various uses such as a dental graft material, an orthopedic graft material, a scaffold, drug delivery system and the likes.

Description

BACKGROUND OF THE INVENTION 1. Field of the Invention [0001] The present invention relates to a method for producing a hyaluronic acid derivative sponge, and a hyaluronic acid derivative sponge produced thereby,

The present invention relates to a method for producing a sponge of a hyaluronic acid derivative and a hyaluronic acid derivative sponge produced thereby, and more particularly, to a method for producing a hyaluronic acid derivative sponge by a method of producing a hyaluronic acid derivative by a lyophilization method, And a hyaluronic acid derivative sponge produced by the method. The present invention also relates to a method for producing a hyaluronic acid derivative sponge, which is capable of providing a hyaluronic acid derivative sponge that is superior in toxicity,

Hyaluronic acid is a type of polysaccharide that is linearly linked with repeating units composed of N-acetyl-glucosamine and D-glucuronic acid. It is a biopolymer substance and is first separated from the liquid filling the animal's eye [Meyer K. et (synovial fluid, pleural fluid, skin, rooster, and the like) of animal placenta, joints since it is known to be present in many cases, Streptococcus microorganisms Streptococcus equi, Streptococcus zooepidemecus, etc. are also produced.

Hyaluronic acid is excellent in biocompatibility and has high viscoelastic properties in a solution state, and is widely used not only for cosmetic applications such as cosmetic additives but also for a variety of medicines such as eye surgery aids, joint function improvers, drug delivery materials and eyedrops. However, hyaluronic acid itself is easily decomposed in vivo or in acid or alkali conditions and its use is limited. Therefore, much efforts have been made to develop structurally stable hyaluronic acid derivatives through cross-linking or attaching functional groups of hyaluronic acid (Laurent, TC, et al., Portland Press Ltd, London, 1998 ).

In addition, as an attempt to modify the physicochemical and biological properties of hyaluronic acid and to apply it to the above-mentioned various applications, there have been proposed a support, a carrier for a drug delivery system, a wound covering material, a dental material, and a medical material It is known to produce hyaluronic acid in the form of a sponge.

As a method of producing a hyaluronic acid sponge, it is known that natural hyaluronic acid is easily dissolved in water and thus it is difficult to obtain a sponge form having good properties. Therefore, it is known to crosslink hyaluronic acid so as to maintain the sponge form after drying by changing hyaluronic acid insoluble have. Various cross-linking agents have been used to prepare the crosslinked hyaluronic acid. Among them, typical examples thereof include divinyl sulfone (U.S. Patent No. 4582865), formaldehyde (Japanese Patent Laid-open No. 60-130601), carbodiimide Patent No. 94/2517), epoxide (US Patent Publication No. 2011-0268706) crosslinking agent, and the like have been proposed.

However, the porous hyaluronic acid sponge proposed in the above-mentioned U.S. Patent Publication No. 2011-0268706 has a problem in that it is difficult to repeatedly reproduce a sponge having a good shape in the process of producing a sponge by expanding the crosslinked hyaluronic acid solution unpredictably have. In addition, when divinylsulfone or aldehydes are used as a crosslinking agent, the cytotoxicity caused by unreacted residues may be a problem, and there is a limitation in use, and there is a problem in that water-soluble carbodiimides which are used for crosslinking biomaterials having amino acid components Imid does not exhibit toxicity in the body, but it has a disadvantage that it must be used in excess to show a crosslinking effect and be expensive.

On the other hand, US Patent No. 4970298 discloses a method for producing a collagen sponge, which is a biodegradable matrix, for use in applications similar to the above-mentioned hyaluronic acid sponge. The hyaluronic acid derivative sponge has a lower mechanical strength, The performance as a medical material for culture and transplantation of cells and tissues is rather low.

Accordingly, it is an object of the present invention to provide a hyaluronic acid derivative sponge which can be easily produced in the form of a sponge with good properties, has excellent mechanical strength, is toxic and is economical and has excellent biodegradation control ability And a hyaluronic acid derivative sponge produced by the method.

The present invention relates to a method for producing a hyaluronic acid derivative, comprising the steps of: i) obtaining an hyaluronic acid derivative by adding an epoxide crosslinking agent having two or more epoxy reactive groups to an aqueous solution of hyaluronic acid in sodium hydroxide; ii) washing the crosslinked hyaluronic acid derivative with ultrapure water; iii) pulverizing the washed hyaluronic acid derivative; iv) lyophilizing the pulverized hyaluronic acid derivative to obtain a powder; v) re-swelling the hyaluronic acid derivative powder with ultrapure water to form a desired shape; And vi) finally lyophilizing the re-swelled hyaluronic acid derivative. The hyaluronic acid derivative sponge and the hyaluronic acid derivative sponge produced thereby are provided.

The hyaluronic acid is characterized by being hyaluronic acid, hyaluronic acid salt, or a mixture of hyaluronic acid and hyaluronic acid salt.

Wherein the hyaluronic acid salt is at least one selected from the group consisting of sodium hyaluronate, potassium hyaluronate, calcium hyaluronate, magnesium hyaluronate, zinc hyaluronate, cobalt hyaluronate and tetrabutylammonium hyaluronate.

The epoxide crosslinking agent may be 1,4-butanediol diglycidyl ether (BDDE), ethylene glycol diglycidyl ether (EGDGE), 1,6-hexanediol di Propylene glycol diglycidyl ether, 1,6-hexanediol diglycidyl ether, propylene glycol diglycidyl ether, polypropylene glycol diglycidyl ether, Poly (tetramethylene glycol) diglycidyl ether, neopentyl glycol diglycidyl ether, polyglycerol polyglycidyl ether, diglycerol polyglycidyl ether, diglycerol polyglycidyl ether, glycerol polyglycidyl ether, trimethylpropane polyglycidyl ether, 1,2- (bis (2,3- One selected from the group consisting of 1,2- (bis (2,3-epoxypropoxy) ethylene), pentaerythritol polyglycidyl ether, and sorbitol polyglycidyl ether Or more.

The cross-linking density of the hyaluronic acid derivative is 1 mol% to 100 mol%.

The weight ratio of the epoxide crosslinking agent is characterized in that 1 to 100 parts by weight based on 100 parts by weight of the repeating unit of hyaluronic acid.

The hyaluronic acid derivative is obtained by adding a polymer capable of forming an ether covalent bond with hyaluronic acid to the hyaluronic acid or hyaluronate solution in the step i), and reacting the resultant with an epoxide crosslinking agent.

The polymer capable of forming an ether covalent bond with the hyaluronic acid is selected from the group consisting of collagen, alginic acid, chitosan, heparin, gelatin, elastin, fibrin, laminin, fibronectin, proteoglycan, heparan sulfate, chondroitin sulfate, dermatan sulfate and keratan sulfate And at least one selected from the group consisting of

The polymer capable of forming an ether covalent bond with hyaluronic acid is selected from the group consisting of polyethylene oxide, polyvinyl alcohol, polypropylene oxide, polyethylene oxide-polypropylene oxide copolymer, polyethylene oxide-polylactic acid copolymer, polyethylene oxide-polylactic glycolic acid Polyolefins such as polyethylene oxide-polycaprolactone copolymer, polybutylene oxide, polyoxyethylene alkyl ethers, polyoxy-ethylene-co-ester oil derivatives, polyoxyethylene sorbitan fatty acid esters and polyoxyethylene stearates And at least one selected from the group consisting of

Characterized in that natural hyaluronic acid is added in step (v).

The present invention provides a hyaluronic acid derivative sponge produced by the above production method.

Wherein the hyaluronic acid derivative sponge further comprises an antibacterial or anti-inflammatory natural substance.

The antimicrobial or anti-inflammatory natural substance may be selected from the group consisting of rosemary, clove, white porcelain, turmeric, green tea, black soybean seed, rose petal, peony root, bellflower, bean sprouts, colored barley seed, camellia petal, buckwheat, grapefruit, licorice, And at least one selected from the group consisting of gold, cinnamon, grasses, brambles, rhododendron, cherry, forsythia, pepper leaves, mint, bananas, mulberry, saururus, pine, .

The present invention provides a dental implant, an orthopedic implant, a cell support, or a drug delivery system comprising the hyaluronic acid derivative sponge.

According to the present invention, a hyaluronic acid derivative sponge having excellent mechanical strength, no toxicity, and excellent biodegradation control ability can be obtained by easily and repeatedly producing a hyaluronic acid derivative with a good shape sponge.

In addition, the hyaluronic acid derivative sponge produced by the present invention has excellent biocompatibility, biodegradation control ability, and mechanical strength, and can be used for various applications such as dental implant, orthopedic implant, cell support, and drug delivery system.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a graph showing the compressive strength of a hyaluronic acid derivative sponge manufactured according to Example 1 of the present invention and a commercially available collagen sponge in a dry state and a swollen state (Test Example 1).
2 is a graph showing absorption capacity of a hyaluronic acid derivative sponge produced according to Example 1 (Experimental Group 2) of the present invention (Test Example 2).
FIG. 3 is a graph showing the results of an animal experiment in which the degree of bone regeneration in Example 1 (Experimental Group 2) of the present invention was evaluated. The experimental group 2 using the hyaluronic acid derivative sponge of the present invention and the bone in the control group (Test Example 3).

The present invention provides a method for producing a hyaluronic acid derivative sponge and a hyaluronic acid derivative sponge produced thereby.

Hereinafter, the present invention will be described in detail.

Specifically, the present invention relates to a process for producing a hyaluronic acid by cross-linking hyaluronic acid using an epoxide crosslinking agent having two or more epoxy reactive groups and then easily producing a crosslinked hyaluronic acid derivative in a sponge form having good properties by freeze- will be.

The present invention relates to a method for producing a hyaluronic acid derivative, comprising the steps of: i) obtaining an hyaluronic acid derivative by adding an epoxide crosslinking agent having two or more epoxy reactive groups to an aqueous solution of hyaluronic acid in sodium hydroxide; ii) washing the crosslinked hyaluronic acid derivative with ultrapure water; iii) pulverizing the washed hyaluronic acid derivative; iv) lyophilizing the pulverized hyaluronic acid derivative to obtain a powder; v) re-swelling the hyaluronic acid derivative powder with ultrapure water to form a desired shape; And vi) finally lyophilizing the re-swelled hyaluronic acid derivative. The hyaluronic acid derivative sponge and the hyaluronic acid derivative sponge produced thereby are provided.

The term "hyaluronic acid" as used herein means hyaluronic acid, hyaluronic acid salt, or a mixture of hyaluronic acid and hyaluronic acid salt. The hyaluronic acid salt is characterized in that it is at least one selected from the group consisting of sodium hyaluronate, potassium hyaluronate, calcium hyaluronate, magnesium hyaluronate, zinc hyaluronate, cobalt hyaluronate and tetrabutylammonium hyaluronate, Sodium hyaluronate is used, but is not limited thereto.

When the hyaluronic acid in step i) is dissolved in an aqueous solution of sodium hydroxide, the concentration of the aqueous solution of sodium hydroxide is preferably 0.05 N to 1.0 N, more preferably 0.1 N to 0.5 N. If the concentration of sodium hydroxide aqueous solution is less than 0.05 N, the water absorption amount becomes high and it is difficult to grind and refine. If the concentration exceeds 1.0 N, the physical property is lowered. Therefore, the concentration of aqueous sodium hydroxide solution is adjusted to 0.05 N to 1.0 N.

In order to obtain the hyaluronic acid derivative of the step i), the hyaluronic acid aqueous solution to which the epoxide crosslinking agent is added is reacted at 20 to 40 for 12 to 36 hours.

The cross-linking density of the hyaluronic acid derivative in the step i) may be 1 mol% to 100 mol%, and more preferably 5 mol% to 50 mol%. When the crosslinking density is less than 1 mol%, the physical properties are lowered and the water absorption capacity is increased, which makes purification difficult. When the crosslinking density is more than 100 mol%, the possibility of a large amount of crosslinking agent remaining is increased.

The epoxide crosslinking agent used in the present invention is 1,4-butanediol diglycidyl ether (BDDE), ethylene glycol diglycidyl ether (EGDGE), 1,6-hexanediol diglycidyl ether Hexanediol diglycidyl ether, propylene glycol diglycidyl ether, polypropylene glycol diglycidyl ether, and the like. Poly (tetramethylene glycol diglycidyl ether), neopentyl glycol diglycidyl ether, polyglycerol polyglycidyl ether, diglycerol polyglycidyl ether, polyglycerol polyglycidyl ether, Diglycerol polyglycidyl ether, glycerol polyglycidyl ether, trimethylpropane polyglycidyl ether, and the like. (1,2- (bis (2,3-epoxypropoxy) ethylene), pentaerythritol polyglycidyl ether, or sorbitol And sorbitol polyglycidyl ether. More preferably, 1,4-butanediol diglycidyl ether is used, but the present invention is not limited thereto.

The crosslinking agent containing two or more epoxy functional groups reacts with an alcohol group of hyaluronic acid to form an ether bond, thereby crosslinking.

The weight ratio of the hyaluronic acid epoxide crosslinking agent is 1 to 100 parts by weight, more preferably 5 to 50 parts by weight, based on 100 parts by weight of the repeating unit of the hyaluronic acid. When the weight ratio of the hyaluronic acid epoxide crosslinking agent is less than 1 part by weight, the physical properties are lowered and the water absorption capacity is increased, which makes purification difficult. When the weight ratio exceeds 100 parts by weight, the possibility that a large amount of crosslinking agent remains is increased.

In the present invention, the hyaluronic acid derivative may be obtained by adding a hyaluronic acid or a hyaluronic acid salt alone, or a polymer capable of forming an ether covalent bond with hyaluronic acid or a hyaluronic acid or a hyaluronic acid salt solution by an epoxide crosslinking agent.

The polymer capable of forming an ether covalent bond with the hyaluronic acid may be selected from the group consisting of natural high molecular weight collagen, alginic acid, chitosan, heparin, gelatin, elastin, fibrin, laminin, fibronectin, proteoglycan, heparan sulfate, chondroitin sulfate, And lanthanum sulfate.

The polymer capable of forming an ether covalent bond with hyaluronic acid may be a synthetic high molecular weight polyethylene oxide, polyvinyl alcohol, polypropylene oxide, polyethylene oxide-polypropylene oxide copolymer, polyethylene oxide-poly Polyoxyethylene alkylphenols, polyoxyethylene alkylphenols, polyoxyethylene alkylphenols, polyoxyethylene alkylphenols, polyoxyethylene alkylphenols, polyoxyethylene alkylphenols, polyoxyethylene alkylphenols, polyoxyethylene alkylphenols, polyoxyethylene alkylphenols, Fatty acid esters, polyoxyethylene stearates, and the like.

The process in which hyaluronic acid of the present invention is obtained as a hyaluronic acid derivative by an epoxide crosslinking agent is shown in Reaction Scheme 1 below.

(Scheme 1)

The size of the pulverized hyaluronic acid derivative in the process of pulverizing the hyaluronic acid derivative in the step iii) is preferably 100 μm to 500 μm, but it may be pulverized into other sizes as required.

In the process of re-swelling the hyaluronic acid derivative powder of step (v) with ultra-pure water, the hyaluronic acid derivative powder is preferably swelled to 1% to 10%, more preferably 3% to 5% of the ultrapure water mass. When swelling less than 1%, a sponge with good properties is not formed well after lyophilization, and when it is swollen to 10% or more, the hyaluronic acid derivative powder does not swell sufficiently and there is a problem that the sponge is broken well. Hyaluronic acid derivative powder, It should be adjusted within the range of 1% to 10%.

In addition, natural hyaluronic acid may be added during the re-swelling process of step (v) in order to improve the mechanical strength, and the concentration of the natural hyaluronic acid to be added is not particularly limited. However, Is lowered, it is preferably 0.01% to 5% of the ultrapure water mass.

In the present invention, the freeze-drying is performed for about 24 hours in a freezer at -20 ° C to -100 ° C and then again for 48 hours or more at a temperature of -50 ° C or lower and a pressure of 5 mTorr to 50 mTorr to obtain a sponge Can be easily reproduced.

The hyaluronic acid derivative sponge produced by the present invention has high adherence and has good biocompatibility because it binds well to living tissue. In addition, since it has stability against decomposition enzyme, it maintains its volume in the body for a certain period of time, and after it is completely decomposed and absorbed into living body, it has excellent ability to regulate biodegradation.

Due to these properties, the hyaluronic acid derivative sponge produced by the present invention can be used for various applications such as dental implant, orthopedic implant, cell scaffold, and drug delivery system.

The hyaluronic acid derivative sponge of the present invention may further include an antibacterial or anti-inflammatory natural substance. The antibacterial or anti-inflammatory natural substance may be selected from the group consisting of rosemary, clove, white porcelain, turmeric, green tea, black soybean seed, rose petal, Bamboo leaves, peppermint, mint, bamboo, mulberry, bamboo, pine, bamboo, bamboo, bamboo, bamboo, Mugwort, persimmon, Yoshino cherry tree, Sasae, or both stem extracts.

Hereinafter, embodiments of the present invention will be described in detail.

(Example)

Example  1: Production of hyaluronic acid derivative sponge

A hyaluronic acid solution in which hyaluronic acid was dissolved in 0.25N NaOH solution at 10 wt% was prepared in each of three reactors. 30 parts by weight, 40 parts by weight and 50 parts by weight of 1,4-butanediol diglycidyl ether (BDDE), which is a crosslinking agent, were added to 100 parts by weight of hyaluronic acid in the solution, For 30 hours. The reactants were washed several times with ultrapure water to remove unreacted materials, and the washed product was pulverized to adjust the particle size from 100 μm to 500 μm. The pulverized reaction product was lyophilized at -50 ° C and 5 mTorr for 72 hours, and the powder obtained after lyophilization was re-swollen to ultrapure water at a 3% weight ratio. The swollen reactant was placed in a mold of the desired shape and lyophilized to prepare a hyaluronic acid derivative sponge. The hyaluronic acid derivative sponge was finally made into Φ 8 mm * 25 mm. The hyaluronic acid derivative sponge prepared in order to carry out the experiment of the present invention was named Experimental group 1 (30 parts by weight), Experimental group 2 (40 parts by weight) and Experimental group 3 (50 parts by weight) according to the ratio of crosslinking agent.

Test Example  1. Measurement of mechanical strength of hyaluronic acid derivative sponge

The hyaluronic acid derivative sponge prepared in Example 1 and the collagen sponge licensed from the Korean Food and Drug Administration were cut into a shape of? 8 mm * 5 mm, and the sponge was compressed to a height of 1 mm using a rheometer The compressive strength was measured. The compressive strength of the sponge was measured by using a dry sponge and a PBS (phosphate buffered saline) for 1 hour.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a graph comparing the compressive strengths of a hyaluronic acid derivative sponge and a collagen sponge of the present invention in a dry state and a swollen state, respectively. The compressive strength of the hyaluronic acid derivative sponge of the present invention is compared with the compressive strength of a collagen sponge, It was confirmed that the hyaluronic acid derivative sponge of the present invention is superior in mechanical strength to commercialized collagen sponge.

Test Example  2. Hyaluronic acid derivative sponge Absorption capacity

To measure the absorption capacity of the experimental group 2 prepared in Example 1, the mass of the hyaluronic acid derivative sponge was measured and 10 ml of physiological saline was added. After allowing to stand at 37 DEG C for 72 hours, the mass of the hyaluronic acid derivative sponge absorbed with physiological saline was measured, and the absorption capacity of the sponge was calculated by the following formula (1).

Equation 1

Absorption capacity (%) = 100 X [mass of swollen sponge] / [initial mass of sponge]

Fig. 2 shows the absorption ability of the hyaluronic acid derivative sponge. The hyaluronic acid derivative sponge of the present invention exhibited the ability to absorb moisture, which is at least 30 times its volume, while retaining its shape.

Test Example  3. Hyaluronic acid derivative sponge Bone regeneration  Degree evaluation

In order to confirm the bone regeneration effect of the experimental group 2 prepared in Example 1, 8-week old SD rats were used as model animals. After anesthetizing the animal, the skin at the center of the forehead was incised about 4 cm, and the periosteum was incised. Using a dental drill with a diameter of 8 mm, paying attention to the damage of the blood vessels passing through the dura mater and the two midbodies, A critical size defect of 8 mm in diameter [J. P Schmitz et al., Clin. Orthop. Relat. Res., 205, 299 (1986)], and the experimental group 2 was shaped into Φ 8 mm * 2 mm to fill the defect, and the periosteum and skin were sutured. In addition, a control group, which was stitched without any treatment on the bone defect part, was also tested. After 12 weeks of feeding, animals were sacrificed and bone defects were collected and evaluated by soft X-ray and tissue staining.

As shown in Fig. 3, the experimental group 2 showed better bone regeneration than the control group. Based on these results, it was confirmed that the hyaluronic acid derivative sponge produced by the present invention can be used as a dental material or orthopedic implant with excellent bone regeneration ability.

Test Example  4. Safety evaluation of hyaluronic acid derivative sponge

For the safety evaluation of the hyaluronic acid derivative sponge prepared in Example 1 of the present invention, a nonclinical toxicity test was conducted in the GLP certification body, and the results are shown in Table 1. [

Test Items Test Methods Test result Cytotoxicity test Performed according to ISO 10993-5 No toxicity Sensitization test Performed according to ISO 10993-10 No sensation Irritation test Performed according to ISO 10993-10 Non-magnetic Acute toxicity test Performed according to ISO 10993-11 No toxicity Pyrogenicity test Performed according to USP <151> Rabbit Pyrogen Test voice Genotoxicity test
(Micronucleus) Performed in accordance with ISO 10993-3 and OECD Toxicology Test Guideline (No. 474) Does not cause micronuclei Genotoxicity test
(Return mutation test) Performed according to ISO 10993-3 and OECD Toxicology Test Guideline (No. 471) Do not mutate Transplantation test Performed according to ISO 10993-6 No irritation Hemolytic test Performed according to ISO 10993-4 No hemolysis

As can be seen from the stability evaluation results of Table 1, the hyaluronic acid derivative sponge produced by the present invention was confirmed to be a safe substance in various indicators of the nonclinical toxicity test.

Claims (14)

i) adding an epoxide crosslinking agent having two or more epoxy reactive groups to an aqueous solution of hyaluronic acid dissolved in sodium hydroxide to obtain a hyaluronic acid derivative;
ii) washing the crosslinked hyaluronic acid derivative with ultrapure water;
iii) pulverizing the washed hyaluronic acid derivative;
iv) lyophilizing the pulverized hyaluronic acid derivative to obtain a powder;
v) re-swelling the hyaluronic acid derivative powder with ultrapure water to form a desired shape; And
vi) finally lyophilizing the reswelled hyaluronic acid derivative. The method according to claim 1, wherein the hyaluronic acid is hyaluronic acid, a hyaluronic acid salt, or a mixture of hyaluronic acid and hyaluronic acid salt. The hyaluronic acid salt according to claim 2, wherein the hyaluronic acid salt is at least one selected from the group consisting of sodium hyaluronate, potassium hyaluronate, calcium hyaluronate, magnesium hyaluronate, zinc hyaluronate, cobalt hyaluronate and tetrabutylammonium hyaluronate By weight based on the weight of the hyaluronic acid derivative sponge. The epoxy resin composition according to claim 1, wherein the epoxide crosslinking agent is 1,4-butanediol diglycidyl ether (BDDE), ethylene glycol diglycidyl ether (EGDGE), 1 Hexylene glycol diglycidyl ether, propylene glycol diglycidyl ether, poly (propylene glycol) diglycidyl ether (propylene glycol diglycidyl ether) ), Poly (tetramethylene glycol diglycidyl ether), neopentyl glycol diglycidyl ether, polyglycerol polyglycidyl ether, di Examples include glycerol polyglycidyl ether, glycerol polyglycidyl ether, trimethylpropane polyglycidyl ether, (bis (2,3-epoxypropoxy) ethylene), pentaerythritol polyglycidyl ether, or sorbitol Wherein the surfactant is at least one selected from the group consisting of sorbitol polyglycidyl ether and polyglycidyl ether. The method according to claim 1, wherein the crosslinking density of the hyaluronic acid derivative is 1 mol% to 100 mol%. The method of claim 1, wherein the weight ratio of the epoxide crosslinking agent is 1 to 100 parts by weight based on 100 parts by weight of the repeating unit of hyaluronic acid. The method of claim 1, wherein the hyaluronic acid derivative is obtained by reacting the hyaluronic acid or the hyaluronic acid salt solution with a polymer capable of forming an ether covalent bond with the hyaluronic acid salt in step i) and reacting with an epoxide crosslinking agent. Method for producing a hyaluronic acid derivative sponge. The method of claim 7, wherein the polymer capable of forming an ether covalent bond with hyaluronic acid is collagen, alginic acid, chitosan, heparin, gelatin, elastin, fibrin, laminin, fibronectin, proteoglycan, heparan sulfate, chondroitin sulfate, dermatan sulfate And keratan sulfate, wherein the hyaluronic acid derivative sponge is selected from the group consisting of keratan sulfate. The method of claim 7, wherein the polymer capable of forming an ether covalent bond with hyaluronic acid is polyethylene oxide, polyvinyl alcohol, polypropylene oxide, polyethylene oxide-polypropylene oxide copolymer, polyethylene oxide-polylactic acid copolymer, polyethylene Oxide-polylactic glycolic acid copolymer, polyethylene oxide-polycaprolactone copolymer, polybutylene oxide, polyoxyethylene alkyl ethers, polyoxy-ethylene castor oil derivatives, polyoxyethylene sorbitan fatty acid esters and polyoxy A method for producing a hyaluronic acid derivative sponge, characterized in that at least one member selected from the group consisting of ethylene stearate. The method of producing a hyaluronic acid derivative sponge according to claim 1, wherein natural hyaluronic acid is added in step (v). Hyaluronic acid derivative sponge prepared by the method of any one of claims 1 to 10. 12. The hyaluronic acid derivative sponge according to claim 11, further comprising an antibacterial or anti-inflammatory natural substance. 13. The method of claim 12, wherein the antimicrobial or anti-inflammatory natural substance is selected from the group consisting of rosemary, clove, white porcelain, turmeric, green tea, black soybean seed roses, rose petal, peony root, bellflower, bean sprouts, Selected from the group consisting of red, white, yellow, white, golden, cinnamon, grass, bramble, rhododendron, oak, forsythia, peppermint, bamboo, mulberry, saururus, pine, mugwort, Wherein the hyaluronic acid derivative sponge is at least one kind of hyaluronic acid derivative sponge. A dental implant, an orthopedic implant, a cell support, or a drug delivery device comprising the hyaluronic acid derivative sponge of claim 11.

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