WO2016141892A1

WO2016141892A1 - Sheet-like cross-linked hyaluronate hydrogel and preparation method thereof

Info

Publication number: WO2016141892A1
Application number: PCT/CN2016/076177
Authority: WO
Inventors: 蒙一纯
Original assignee: 北京蒙博润生物科技有限公司
Priority date: 2015-03-12
Filing date: 2016-03-11
Publication date: 2016-09-15
Also published as: CN104721873A; CN104721873B

Abstract

A sheet-like cross-linked hyaluronate hydrogel and preparation method and use thereof. The sheet-like cross-linked hyaluronate hydrogel comprises a cross-linked hyaluronate hydrogel and a net-shaped fiber, wherein the net-shaped fiber is embedded in the cross-linked hyaluronate hydrogel, and forms a cement-steel structure.

Description

Chip crosslinked hyaluronic acid hydrogel and preparation method thereof

The present application claims priority to Chinese Patent Application No. 201510107032.6, filed on Mar.

Technical field

This invention relates to a sheet crosslinked hyaluronate hydrogel useful as a dressing. The flaky crosslinked hyaluronate hydrogel provides good mechanical properties, moderate ductility, long-term water retention, water vapor transmission rate, liquid absorption, comfort and transparency. The invention further relates to a process and use for the preparation of the flaky crosslinked hyaluronic acid hydrogel.

Background technique

The hydrogel is a gel-like substance having a three-dimensional network structure composed of a natural or synthetic hydrophilic high molecular polymer, and a rich water molecule is dispersed in the three-dimensional network structure. The hydrogel itself contains a certain amount of water and, due to the presence of hydrophilic residues, it can further absorb water until it reaches saturation. The ability of a hydrogel to absorb additional water varies with the specific composition of the hydrogel.

The biological properties of hydrogels are close to loose connective tissue, and show application prospects in the fields of cell growth scaffolds, medical electrodes, biosensors, drug delivery vehicles, contact lenses, stem cell storage, tissue filling, wound healing, etc. It is the role played by hydrogels in wound healing that has attracted the attention of researchers.

In 1962, British scientist Dr. George Winter discovered that a moist environment is conducive to wound healing. The literature shows that hydrogel dressings are moisturizing, provide a moist environment for the wound, and have a water vapor transmission rate and flexibility that relieves pain and does not destroy the wound tissue that has healed when replaced. Recent studies have demonstrated that hydrogels facilitate capillary angiogenesis at the wound site and proliferation and migration of hair follicle epidermal cells.

Excellent hydrogel wound dressings should have the following performances: (1) Good biocompatibility, which is conducive to cell growth and cell products (cytokines) at the wound site. Produce and function; (2) Selective barrier, prevent the invasion of undesirable factors, prevent body fluid loss, and have good water vapor transmission rate; (3) Control and absorb wound exudate; (4) Have good Mechanical properties; (5) long-term maintenance of the humid environment, reduce replacement; (6) close to the wound but not adhered, easy for clinical use.

Depending on the material source of the hydrogel, hydrogels can be classified into synthetic polymeric hydrogels and natural polymeric hydrogels, as well as combinations of the two. Synthetic polymeric hydrogels include, for example, polyethyleneimine hydrogels (CN101980729A), polyacrylic acid hydrogels, polyvinyl alcohol hydrogels (CN101337086A). Natural polymeric hydrogels include, for example, chitosan hydrogels, alginic acid hydrogels (CN102100592B). However, the role of hyaluronate hydrogels in medical applications is only found in the patent (CN1433432A) and a large number of theoretical studies.

Hyaluronic acid is a linear polymeric polysaccharide polymer formed by repeated attachment of a disaccharide unit composed of β-D-N-acetylglucosamine and β-D-glucuronic acid, which naturally exists in the form of a hyaluronate in the body. Hyaluronate is not immunogenic and exhibits excellent biocompatibility after transplantation or injection into living organisms. Therefore, hyaluronate hydrogels and crosslinked hyaluronate hydrogels have been widely used in osteoarthritis, eye surgery, facial wrinkles and filling. See, for example, U.S. Patent No. 4,582, 865, issued to Japanese Patent Application Publication No. No. No. No. No. No. No. No. No. No. No. No. No. No. No. No. No. No. No. No. No. No. No. No. No. Japanese Patent Publication No. 97401 and Japanese Patent Laid-Open No. Hei 9-59303. The hyaluronic acid hydrogel has better biocompatibility than the natural materials such as chitosan and alginic acid described above. Therefore, hyaluronate hydrogels are more commercially valuable.

A large number of reports have reported that hyaluronate hydrogels have a healing effect on wounds. However, although hyaluronate hydrogels have good biocompatibility and thus become a hot spot for scholars, few of them are commercially available. The main reason is that neither the hyaluronate hydrogel nor the crosslinked hyaluronate hydrogel lacks stable mechanical properties and cannot form a sheet-like structure that facilitates clinical operations.

Therefore, it is thought to use a method of preparing a composite material to obtain a hyaluronic acid hydrogel having a certain mechanical strength as a dressing, but the composite material currently used as a dressing is composed of a hyaluronic acid hydrogel, a gas permeable membrane, and a non-woven fabric. Cloth and / or medical adhesive The composite of the multiple layers of the mixture has the disadvantage of poor water vapor transmission rate, transparency and liquid absorption.

For example, CN103432621A discloses an amorphous hyaluronate hydrogel dressing comprising various components such as hyaluronic acid, propylene glycol, glycerin and water, and further coated or soaked on a nonwoven fabric to directly contact the skin. For the treatment of acne, dermatitis and eczema.

Chung-Daw Young et al. (High-strength, ultra-thin and fiber-reinforced pHEMA artificial skin, Biomaterials, 19, (1998), pp. 1745-1752) discloses a fiber-reinforced polymethyl methacrylate-2-hydroxyl Ethyl hydrogel artificial skin with enhanced mechanical strength. However, the saturated water content of the hydrogel (referred to herein as equilibrium water content (EWC), which is 32.4-36.0% by weight) is low, and the absorbed water is almost completely evaporated in the air in 1 hour (see Young et al. , Table 1 and Figure 4). At the same time, it is not disclosed that the hydrogel has liquid absorbability. Therefore, the fiber-reinforced hydrogel is not suitable for use as a desired dressing. This document also does not specifically disclose the preparation of the fiber-reinforced hydrogel.

CN1822866A, citing Young et al. and making improvements in mechanical strength, discloses a fiber-reinforced poly-2-hydroxyethyl methacrylate hydrogel as a material for replacing cartilage-like tissue. However, this patent does not involve dressing applications, nor does it overcome the problems of Young et al.

Accordingly, there is a need in the art for crosslinked hyaluronic acid hydrogel dressings having good mechanical properties, moderate ductility, long term water retention, water vapor transmission rate, liquid absorbency, comfort, and clarity.

Summary of the invention

The technical problem to be solved by the present invention is to provide a sheet-like crosslinked hyaluronate hydrogel which has good mechanical properties, moderate ductility and long-term in addition to good biocompatibility of the hyaluronate itself. Water retention, water vapor transmission rate, liquid absorbency, comfort, and transparency are suitable, for example, for use in dressings for burn wounds having exudates. The invention also provides preparation of the flaky crosslinked hyaluronate hydrogel Method and use.

In order to solve the above problems, the present invention does not employ a conventional lamination method, but by using a reticular fiber as a skeleton, a sheet-like structure in which a reticular fiber is embedded in a crosslinked hyaluronic acid hydrogel is obtained. A hydrogel having a cement-rebar-like structure is provided to provide a sheet-like crosslinked hyaluronate hydrogel having the above properties. The flaky crosslinked hyaluronic acid hydrogel not only retains the excellent characteristics of the crosslinked hyaluronate hydrogel, but also has good mechanical properties, moderate ductility, long-term water retention, water vapor transmission rate, Liquid absorption, comfort and transparency.

The invention therefore provides a flaky crosslinked hyaluronate hydrogel comprising:

a) cross-linked hyaluronate hydrogel; and

b) reticular fibers;

The reticular fibers are embedded in the crosslinked hyaluronic acid hydrogel and form a cement-reinforced structure.

The present invention also provides a method for preparing a sheet-like crosslinked hyaluronate hydrogel comprising:

a) synthesizing hyaluronic acid water into a hyaluronate hydrogel;

b) pressing the hyaluronate hydrogel onto the reticular fibers into a sheet-like hyaluronate hydrogel in which the reticular fibers are embedded;

c) Cross-linking the sheet-like hyaluronate hydrogel.

The present invention also provides a dressing comprising the sheet-like crosslinked hyaluronate hydrogel of the present invention or a sheet prepared by the method for preparing a sheet-like crosslinked hyaluronate hydrogel according to the present invention. Cross-linked hyaluronate hydrogel.

The present invention also provides a mask comprising the sheet-like crosslinked hyaluronate hydrogel of the present invention or a sheet prepared by the method for preparing a sheet-like crosslinked hyaluronate hydrogel according to the present invention. Cross-linked hyaluronate hydrogel.

The present invention also provides a sheet-like crosslinked hyaluronate hydrogel of the present invention or a sheet-like crosslinked hyaluronic acid prepared by the method for preparing a sheet-like crosslinked hyaluronate hydrogel according to the present invention. The use of a saline gel as a dressing, mask or sustained release carrier.

The sheet of the present invention is compared to an amorphous non-crosslinked or crosslinked hyaluronate hydrogel The crosslinked hyaluronic acid hydrogel has reticular fibers embedded therein, thereby significantly improving the overall mechanical properties and comfort as a dressing by using a reticular fiber having significant mechanical properties as a skeleton.

The body of the sheet-like crosslinked hyaluronate hydrogel of the present invention is compared with a composite obtained by coating a non-crosslinked or crosslinked hyaluronate hydrogel onto a substrate such as a nonwoven fabric. Part of it is water, so when it is used as a dressing, there is no substrate (for example, a nonwoven fabric) as a barrier between the wound and the outside, and thus has a significantly better water vapor transmission rate, liquid absorbability, transparency, and the like.

Compared with hydrogel dressings of other materials, the sheet-like crosslinked hyaluronate hydrogel of the present invention has the significant advantages brought by hyaluronic acid salts, such as biocompatibility, which is more conducive to the growth of cells at the wound site. , migration and healing.

In addition, the water content of the sheet-like crosslinked hyaluronate hydrogel of the present invention can be controlled, for example, between 20 and 80%, making it suitable for various wounds having different degrees of damage and different levels of exudate. It can fully absorb the tissue damage and seep the liquid. The present invention can also obtain a sheet-shaped crosslinked hyaluronate hydrogel having different mechanical properties (such as elastic modulus, viscous modulus, mechanical strength, toughness) by adjusting the degree of crosslinking (for example, 0.5 to 2%) and fibers. To suit different applications.

DRAWINGS

The specific embodiments of the present invention are described in further detail below with reference to the accompanying drawings.

Figure 1 shows a schematic of different reticular fibers.

2 shows a schematic view of a strip crosslinked hyaluronic acid gel extruded in the same direction on a mesh fiber by a machine and a round roll, in accordance with one embodiment of the present invention.

detailed description

For a further and better understanding of the present invention, reference will be made to the particular embodiments of the invention and further preferred and/or optional features.

Unless stated in the operating and comparative examples, or where otherwise explicitly indicated, the quantities or ratios of the materials or reaction conditions, material physical properties and/or All numbers of uses are understood to be modified by the wording "about."

Where the feature is disclosed as being related to a particular aspect of the invention (e.g., a product of the invention), such disclosure is also considered to be applied to any other aspect of the invention (e.g., the method of the invention) and the necessary modifications are made.

The various materials, reagents and equipment used in the present invention are commercially available.

Hyaluronate

The hyaluronate mentioned in the present invention includes hyaluronic acid and a hyaluronate such as an alkali metal salt of hyaluronic acid, such as a sodium salt, a potassium salt, a lithium salt or the like of hyaluronic acid, particularly sodium hyaluronate. And hyaluronic acid or hyaluronate having any modification that does not materially affect the use of hyaluronate in the present invention.

In one embodiment, the hyaluronate is prepared by an animal tissue extraction process or a microbial fermentation process, preferably a microbial fermentation process.

In one embodiment, the sodium hyaluronate is an injectable or pharmaceutical grade sodium hyaluronate in accordance with the Pharmacopoeia of the People's Republic of China.

In some embodiments, the hyaluronate has a weight average molecular weight of from 50 to 5 million Daltons, preferably from 80 to 3 million Daltons, more preferably from 100 to 2 million Daltons.

In one embodiment, the hyaluronate has a molecular weight distribution coefficient D of 3.0.

In one embodiment, the hyaluronate is replaced with one selected from the group consisting of chondroitin sulfate, keratan sulfate, and heparin.

It is known in the art that chondroitin sulfate, keratan sulfate or heparin is a polymer similar to hyaluronate. Those skilled in the art will be able to adjust their weight ratios and degree of crosslinking to obtain similar sheet hydrogels without departing from the spirit and scope of the invention.

Cross-linking

In some embodiments, the crosslinking is selected from the group consisting of physical crosslinking, chemical crosslinking, and self-crosslinking.

In some embodiments, physical crosslinking includes crosslinking that occurs between hydroxyl, carbonyl or amino groups of different macromolecules of hyaluronate under the action of nuclear, optical, electrical, magnetic or ultrasonic energy.

In some embodiments, chemical crosslinking includes cross-linking that occurs between the hydroxyl, carbonyl or amino groups of different macromolecules of hyaluronate under the action of a crosslinking agent.

In some embodiments, self-crosslinking refers to self-crosslinking between hydroxyl, carbonyl or amino groups of different macromolecules of hyaluronate in an ionic medium.

In some embodiments, the hyaluronate crosslinks with the crosslinker.

In some embodiments, cross-linking uses a crosslinking agent comprising a compound having a group selected from the group consisting of a diether group, a diepoxy group, a dialdehyde group, a bisamino group, a bisindenyl group, and a diene bond, preferably comprising A crosslinking agent for a compound of a diepoxy group.

In some embodiments, the crosslinking agent is 1,4-butanediol diglycidyl ether (BDDE) or 1,2,7,8-diepoxyoctane.

In one embodiment, the residual amount of crosslinker in the final product does not exceed 2 ppm.

In some embodiments, the crosslinking is carried out under acidic or basic conditions, preferably under basic conditions.

In one embodiment, the basic condition is a pH of from 9 to 14, preferably a pH of from 13 to 14, more preferably a pH of 13.

In some embodiments, the basic condition refers to an alkaline solution which may be an alkali metal hydroxide solution, such as a sodium hydroxide solution.

In some embodiments, the concentration of the sodium hydroxide solution is from 0.5 to 5% by weight, preferably 1% by weight.

In one embodiment, the crosslinking is carried out at a temperature between 0 and 80 ° C, preferably between 10 and 70 ° C, more preferably between 40 and 60 ° C, most preferably between 50 ° C.

In one embodiment, the reaction time for crosslinking is at least 1.5 to 3 hours, preferably 2 hours.

In some embodiments, the cross-linking is performed by forming a hyaluronate hydrogel in an alkaline solution with a cross-linking agent, and then compressing the hyaluronate hydrogel onto the reticular fibers into a sheet-like transparent The acid salt hydrogel is then carried out.

In some embodiments, the degree of crosslinking is between 0.4 and 2%.

Reticulated fiber

In some embodiments, the reticular fibers are comprised of a synthetic material, a natural material, or a combination thereof.

In some embodiments, the synthetic material is a chemically synthetic fibrous material including, but not limited to, polyamide, polypropylene, polyester, and polyvinylidene chloride.

In some embodiments, the natural material is a fibrous material that is grown in nature including, but not limited to, silk, noodles, hemp, and hair.

In some embodiments, the reticular fibers are transparent, translucent, or opaque.

In some embodiments, the reticular fibers are comprised of a degradable or non-degradable material.

In some embodiments, the reticular fibers are biologically evaluated to produce no hazardous materials.

In some embodiments, the reticular fibers are comprised of a material that is biodegradable and does not produce any harmful substances in the body.

In one embodiment, the reticular fibers are absorbable reticular fibers.

In some embodiments, the method of forming the reticular fibers includes molding and braiding.

In some embodiments, the fiber course of the reticular fibers can include warp and weft directions, diagonal directions, and loops (Fig. 1).

In some embodiments, the reticular fibers have a fiber diameter of no more than 2 mm, no more than 1.5 mm, no more than 1 mm, and no more than 0.5 mm.

In some embodiments, the reticular fibers have a pore diameter of from 0.5 mm to 30 mm, preferably from 1 to 30 mm.

In some embodiments, the reticular fibers have a line thickness of 20D.

In some embodiments, the thickness of the reticular fibers is similar to the line thickness.

In one embodiment, the reticular fiber is a polyamide diamond mesh of size 14A having a basis weight of 18 GSM and a yarn of 20D.

In some embodiments, the area of the reticular fibers in the present invention is such that the thickness of the sheet-like crosslinked hyaluronate hydrogel having a substantially uniform thickness formed on the reticular fibers by the hyaluronate hydrogel is 0.01 -3 cm, preferably 0.01-2 cm, more preferably 0.03-2 cm Area.

In some embodiments, the weight ratio of the reticular fibers is no more than 10% by weight based on the weight of the sheet-like crosslinked hyaluronate hydrogel.

In one embodiment, the volume of the reticular fibers is substantially not increased by the absorption of liquid.

Flake crosslinked hyaluronate hydrogel

In some embodiments, the reticular fibers are embedded in a crosslinked hyaluronate hydrogel, and the formation of a cement-reinforced structure is achieved by subjecting a mixture of hyaluronate hydrogel to reticular fibers to undergo crosslinking.

In some embodiments, the sheet crosslinked hyaluronate hydrogel comprises 7-12% by weight, preferably 9-10% by weight, based on the flaky crosslinked hyaluronate hydrogel.

In one embodiment, the flaky crosslinked hyaluronate hydrogel further comprises phosphate-physiological saline.

In one embodiment, the phosphate-physiological saline is present at 20-80% of 100% swelling.

In one embodiment, the flaky crosslinked hyaluronate hydrogel further comprises an additive including, but not limited to, a bacteriostat, an anesthetic, and an epidermal growth factor.

In one embodiment, the flaky crosslinked hyaluronate hydrogel further comprises a bacteriostatic agent.

In one embodiment, the bacteriostatic agent is cerium nitrate, preferably cerium nitrate hexahydrate, and in some cases it is present in an amount from 0.1 to 5%.

In one embodiment, the phosphate-physiological saline contains 0.1-5% cerium nitrate hexahydrate.

In one embodiment, the crosslinked hyaluronate hydrogel is obtained by subjecting the hyaluronate hydrogel to cross-linking.

In some embodiments, the sheet crosslinked hyaluronate hydrogel has a thickness of from 0.01 to 3 cm, preferably from 0.01 to 2 cm, more preferably from 0.03 to 2 cm.

In one embodiment, the sheet crosslinked hyaluronate hydrogel of the present invention is a heat source free and sterilizable obtained by quality control methods and sterilization methods known in the art. of.

Preparation

In some embodiments, the reagents and materials used in the methods of the invention are all commercially available products.

In one embodiment, the reticular fibers are embedded in a crosslinked hyaluronate hydrogel by a pressure infusion method or a composite superposition method.

In one embodiment, the hyaluronate hydrogel is pressed against the two sides of the reticulated fibers from the opposite direction, thereby obtaining a composite material in which the reticulated fibers are embedded in the hyaluronic acid hydrogel. This composite material has a better hydrogel distribution than the composite obtained by directly pressing the hyaluronate hydrogel onto the reticular fibers.

In one embodiment, the method of the invention further comprises adjusting the pH of the crosslinked hyaluronate hydrogel to from 3 to 10, preferably from 6 to 8, with an acidic material.

In some embodiments, the hyaluronate hydrogel is compressed at intervals, such as 10 minutes, during the cross-linking process to remove the generated bubbles and maintain the thickness.

In some embodiments, during the crosslinking process, the hyaluronic acid hydrogel is pressed using a mechanical pressurization method, for example, using a stainless steel round stick to repeatedly roll the hydrogel over the reticular fibers, or the field may be used. Other pressurization methods are known in which the hydrogel is flatly fitted into the plane of the reticular fibers.

In one embodiment, after the end of the crosslinking, pressing can be continued to further remove air bubbles.

In some embodiments, the method of the present invention further comprises drying the sheet-like crosslinked hyaluronate hydrogel in a drying cabinet (for example, 10 to 60 ° C), for example, for 3 hours, after which water or a component containing, for example, bacteriostatic An aqueous solution of the agent (e.g., 0.5 to 5% by weight of cerium nitrate) is rehydrated.

In some embodiments, the method of the present invention further comprises an additional step of adding water such that the crosslinked hyaluronate hydrogel comprises no more than 99.9 weight based on the weight of the flaky crosslinked hyaluronate hydrogel after the additional water addition step Water by weight, no more than 95% by weight, no more than 90% by weight, no more than 85% by weight or no more than 80% by weight of water.

In some embodiments, the method of the present invention further comprises cutting the hydrogel or rehydrated hydrogel of the present invention into a fixed shape, such as a rectangular shape, and sealing the package.

In some embodiments, the method of the present invention further comprises sterilizing the flaky crosslinked hyaluronate hydrogel by, for example, moist heat sterilization (e.g., at 121 ° C for 15 minutes or 115 ° C for 30 minutes).

In some embodiments, the alkaline solution, the water used to rinse the hydrogel after the end of crosslinking, the water or aqueous solution for rehydration, and/or the final product may contain sodium chloride, preferably 0.75-0.95 by weight. % sodium chloride.

In one embodiment, the invention provides a method for preparing a sheet-like crosslinked hyaluronate hydrogel comprising:

a) laying the reticular fibers on the plane of the stainless steel, and uniformly mixing and superimposing the sodium hyaluronate added with the aqueous solution of the cross-linking agent on the reticular fibers, followed by rolling and pressing to make the sodium hyaluronate hydrogel pass through the net. Fibrous fiber

b) cross-linking at 10-70 ° C for 2 hours to make the reticular fibers and sodium hyaluronate hydrogel integrated, during which the rolling pressure is applied every 10 minutes to drive out the generated bubbles and make the final thickness 0.01-2mm;

c) rinsing with running water for 2 hours after 2 hours to elute residual crosslinker;

d) drying the hydrogel and hydrating it non-saturated with phosphate-physiological saline so that the degree of swelling is 20-80% of the degree of saturated swelling;

e) The hydrated hydrogel is cut into the required area, sealed in a heat resistant plastic bag, and subjected to moist heat sterilization.

(1) selecting reticular fibers;

(2) Pre-prepared sodium hyaluronate hydrogel: sodium hyaluronate is a pharmaceutical grade or injection grade prescribed by the Pharmacopoeia, and has a molecular weight of 50-500 million Daltons;

(3) Embedding the reticular fibers with the sodium hyaluronate hydrogel: the reticular fibers are laid on a stainless steel platform, and the uniformly mixed sodium hyaluronate hydrogel is extruded through a self-made spiral roller to form 10mm diameter strip hydrogel, the strip hydrogel is placed in the same direction in the upper middle of the reticular fiber, and the strip of hydrogel is repeatedly rolled and squeezed with a stainless steel round stick. Passing through the mesh of the reticular fibers, so that the strips of hydrogel are flatly fitted to the reticular fibers;

(4) Forming of the sheet rubber: placing the chimeric hydrogel in an environment with a humidity greater than 50% and 10-70 ° C for not less than two hours, in which the stainless steel round stick is repeatedly rolled and extruded. Gel to reduce the generation of bubbles;

(5) Washing: rinse with flowing pure water at room temperature for not less than 2 hours to remove residual cross-linking agent;

(6) drying: drying the hydrogel in a vacuum drying oven (10-60 ° C) for 3 hours;

(7) Hydration: The dried hydrogel is immersed in 0.5 mol of PBS to hydrate the hydrogel to 20-80% of the 100% swelling degree. In this step, PBS may contain 0.5-5% lanthanum nitrate as a suppression. Bacterial agent

(8) Packaging: The hydrated hydrogel was cut into a rectangular shape and molded into a polyester bag.

(9) Sterilization: The packaged hydrogel was moist heat sterilized at 121 ° C for 15 minutes.

Measurement methods

The hyaluronate salt of the sheet-like crosslinked hyaluronate hydrogel of the present invention can be measured according to the published YY/T 0962-2014 (People's Republic of China Pharmaceutical Industry Standard: Crosslinked Sodium Hyaluronate Gel for Plastic Surgery) content.

The sheet cross-linking transparency of the present invention can be measured according to Section 3.2 of the published YY/T 0471.2-2004 (People's Republic of China Pharmaceutical Industry Standard: Contact Surface Wound Dressing Test Method Part 2: Vapor Transmission Rate of Breathable Membrane Dressing) The water vapor transmission rate of the acid salt hydrogel.

The liquid absorption of the sheet-like crosslinked hyaluronate hydrogel of the present invention can be measured according to Section 3.8 of YY/T 0471.1-2004 (People's Republic of China Pharmaceutical Industry Standard: Contact Surface Wound Dressing Test Method Part 1: Liquid Absorption) Sex.

The comfort of the sheet-like crosslinked hyaluronate hydrogel of the present invention can be measured according to the published YY/T 0471.4-2004 (People's Republic of China Pharmaceutical Industry Standard: Contact Surface Wound Dressing Test Method Part 4: Comfort). Comfort refers to the ability of a wound dressing to adapt to the shape and movement of the human body, which can be expressed quantitatively as stretchability (wound dressing) The force required to stretch to a given stretch range and permanent deformation (the length that is increased after stretching and relaxing the sample, expressed as a percentage of the original length).

The long-term water retention of the sheet-like crosslinked hyaluronate hydrogel of the present invention can be measured by placing the hydrogel in excess water under ambient conditions so that the hydrogel sufficiently absorbs water for 30 minutes, and then it is taken out and allowed to stand. There is essentially no liquid leaving the hydrogel. The hydrogel was then weighed and allowed to stand in an incubator at 25 ° C and 10-30% humidity for a period of time, further weighed at different time points to observe the corresponding water loss.

The transparency of the sheet-like crosslinked hyaluronate hydrogel of the present invention can be evaluated by visual observation.

Product Usage

The sheet-like crosslinked hyaluronate hydrogel of the present invention can be suitably used for various purposes by adjusting the properties such as water content, additives and the like.

In some embodiments, the flaky crosslinked hyaluronate hydrogel of the present invention can be used for skin wounds, such as burns, ulcers, or after laser removal of facial skin dark spots. The sheet-like crosslinked hyaluronate hydrogel of the present invention can be advantageously used as a dressing for clinical use in various skin wounds, including burns, ulcers, such as burn wounds, particularly burn wounds having high levels of exudate. Absorb exudate there and maintain a moist wound environment. The sheet-like crosslinked hyaluronate hydrogel of the present invention can also be used as a dressing for wounds having low or medium levels of exudate.

In one embodiment, the sheet-like crosslinked hyaluronate hydrogel of the present invention can be used as a barrier material for adhesion between organ tissues after surgery.

In one embodiment, the sheet-like crosslinked hyaluronate hydrogel of the present invention can be applied to the field of tissue engineering, for example, for preparing a stem cell culture scaffold or as a substrate for artificial skin.

In one embodiment, the sheet-like crosslinked hyaluronate hydrogel of the present invention can be applied to the field of cosmetics, for example, for preparing a mask.

Example

The invention is further illustrated by the following examples, which are not intended to limit the invention.

The sodium hyaluronate (HA) used in the following examples was purchased from Huaxi Furida Biomedical Co., Ltd., and 1,4-butanediol diglycidyl ether (BDDE, purity ≥ 95%) was purchased from Sigma.

Example 1

10 grams of sodium hyaluronate (molecular weight of about 3 million Daltons) was placed in a 250 ml beaker. In a 250 ml beaker, 500 μl of BDDE was dissolved in 40 ml of 1% sodium hydroxide. The BDDE-mixed lye was poured into a beaker containing sodium hyaluronate, and after thorough mixing, the mixture was charged into a machine as shown in Fig. 2, thereby being extruded into a reticular fiber (polyester). A strip of hydrogel on top. The strip-shaped hydrogel was repeatedly rolled and rolled with a stainless steel round stick so that the sheet-like hydrogel after extrusion had a thickness of 1 mm. The sheet-like hydrogel was placed flat on the bottom of a flat-bottom stainless steel tray and placed in a 50 ° C water bath for 2 hours while the sheet-like hydrogel was taken out during the water bath and rolled four times. The tray was taken out and rinsed with purified water for 2 hours. After draining, the tray was placed in a vacuum oven at 50 ° C for 3 hours. Tablet cross-linked hydrogel using phosphate-physiological saline (0.276 g Na ₂ HPO ₄ ·2H ₂ O in 1000 ml water, 0.0395 g NaH ₂ PO ₄ .·4H ₂ O and 8.476 g NaCl; pH 7.3) Soak for 3 hours to make the degree of swelling 50%. The sheet-like crosslinked sodium hyaluronate hydrogel was cut into a 30 mm × 50 mm rectangle and sealed in a polyester bag. The packaged sheet-like crosslinked sodium hyaluronate hydrogel was sterilized by moist heat at 121 ° C for 15 minutes.

Example 2

500 μl of BDDE in Example 1 was replaced with 1000 μl of 1,2,7,8-diepoxyoctane, and the rest were the same as in Example 1. The resulting sheet-like crosslinked sodium hyaluronate hydrogel has greater elasticity and small viscosity.

Example 3

500 μl of BDDE in Example 1 was replaced with 500 μl of 1,2,7,8-diepoxyoctane, and the rest were the same as in Example 1. The resulting sheet-like crosslinked sodium hyaluronate hydrogel has greater elasticity and small viscosity.

Example 4

The phosphate-physiological saline in Example 1 was replaced with a phosphate-physiological saline containing cerium nitrate (0.276 g of Na ₂ HPO ₄ ·2H ₂ O in 1000 ml of water, 0.0395 g of NaH ₂ PO ₄ .·4H ₂ O and 1 g) Ce(NO ₃ ) ₃ ·6H ₂ O), and the rest were the same as in Example 1. The resulting sheet-like crosslinked sodium hyaluronate hydrogel contained 1% cerium nitrate antibacterial agent. Lanthanum nitrate has low toxicity and good bacteriostatic effect.

Example 5

The 3-hour soaking time of Example 4 was replaced with a 2 hour soaking time so that the degree of swelling was 50%, and the rest were the same as in Example 4. The resulting sheet-like crosslinked sodium hyaluronate hydrogel is suitable for wounds having high exudates.

Example 6

The 3-hour soaking time of Example 4 was replaced with a 4 hour soaking time so that the degree of swelling was 80%, and the rest were the same as in Example 4. The resulting sheet-like crosslinked sodium hyaluronate hydrogel is suitable for wounds having low exudates.

Example 7

The material of the reticular fibers was a long polyester thread, and the rest were the same as in Example 1.

Example 8

The material of the reticular fibers was a transparent polyester plastic thread, and the rest were the same as in Example 1. The resulting sheet-like crosslinked sodium hyaluronate hydrogel is a transparent hydrogel.

Example 9

The material of the reticular fibers was a silk thread, and the rest were the same as in Example 1. The resulting sheet-like crosslinked sodium hyaluronate hydrogel is a biocompatible hydrogel.

Example 10

The material of the reticular fibers was an absorbable suture of the gut, and the rest were the same as in Example 1. The resulting sheet-like crosslinked sodium hyaluronate hydrogel is a tissue absorbable hydrogel.

Example 11. Measurement of the main parameters of the hydrogel of Examples 1-10

(1) Residual agent residue

The BDDE residue of the product of Examples 1-10 was determined to be <1 ppm according to the measurement method described in the industry standard YY/T0962-2014 for Cross-linked Sodium Hyaluronate Gel for Plastic Surgery.

(2) Swelling degree

The degree of swelling of the products of Examples 1-10 was determined to be <25 according to the measurement method described in the industry standard YY/T0962-2014 of "Crosslinked Sodium Hyaluronate Gel for Plastic Surgery".

(3) Intensity

The tensile strength of the products in Examples 1-10 was measured using a material testing machine. The material strength is the value of N recorded when the hydrogel breaks and falls off the reticular fibers during stretching. N>5 was measured.

(4) Transparency

The product obtained in Example 8 was placed in a spectrophotometer cuvette having a thickness of 10 mm, and the transmittance of visible light in the range of 340 to 800 nm was measured at a water transmittance of 100%. The product of Example 8 was measured to have a transmittance of >60%.

(5) Water absorption experiment

10 ml of physiological saline was placed in the container, and the product of Example 1-10 was sealed in a container for 8 hours. The weight (W ₀ ) of the test piece before being placed in the container was compared with the weight (W ₁ ) of the test piece taken out after 8 hours. Absorption amount = (W ₀ ) - (W ₁ ). The amount of water additionally absorbed by the product was measured to be 50% of the degree of swelling before water absorption.

Example 12. Preparation of a sheet-like crosslinked sodium hyaluronate hydrogel using BDDE as a crosslinking agent

Sodium hyaluronate (molecular weight of about 1.2 million Daltons), BDDE and sodium hydroxide solution in an amount described in the "Formulation" section of Table 1 were mixed and stirred to obtain a uniform white granular hydrogel. The hydrogel was divided into two parts, A and B, respectively, and pressed and stretched into a sheet-like hydrogel of about 0.5 mm thick. The reticular fibers (polyamide) were spread on a sheet of hydrogel and the other sheet of hydrogel was symmetrically pressed onto the reticular fibers. The flake-like hydrogel is lightly pressed so that the two sheet-like hydrogels are in contact through the reticular fibers. Subsequently, it was placed in a 50 ° C incubator for 2 h, and then allowed to stand at room temperature for 24 h for use.

The pH was adjusted by soaking the crosslinked hydrogel in a 0.6 mol/ml hydrochloric acid solution for 10 seconds. The pH at various points on the flaky crosslinked hyaluronate hydrogel was examined. If the pH is not in the proper range, the pH can be adjusted by adding an acid solution.

Subsequently, the water described in Table 1 was added to obtain a sheet-like crosslinked sodium hyaluronate hydrogel. The reticular fibers used were polyamide diamond horn meshes of size 20D, 14A.

The sheet-like crosslinked hyaluronate hydrogel was packaged in an aluminum plastic bag and sterilized by damp heat at 111 ° C for 30 min. After the product is allowed to stand for 24 hours, it is packaged to obtain the final product.

Example 13. Measurement of the properties of the sheet-like crosslinked sodium hyaluronate hydrogel of Example 12.

The properties of the different sheet-like crosslinked sodium hyaluronate hydrogels of Example 12 were measured according to the measurement methods described above and are summarized in Table 1.

Table 1. Properties of the flaky crosslinked sodium hyaluronate hydrogel of Example 12.

Example 14. Measurement of long-term water retention of hydrogel after further water absorption (loss of moisture at different time points)

The weight of the sheet-like crosslinked sodium hyaluronate hydrogel of Example 1 of Example 13 which was further absorbed in an incubator at 25 ° C and 10-30% humidity was measured at the time point shown in Table 2, and the result was as follows. As shown in table 2.

The results in Table 2 show that after 120 min, the water content of the hydrogel decreased by only about 19.64%, 20.58% and 18.93% based on the weight of saturated hydrogel after 30 min of water absorption. This indicates that the crosslinked hyaluronate hydrogel of the present invention has excellent water retention capacity.

Overall observations indicate that during the first 2 hours, water was lost at a rate of approximately 5%/30 min, followed by a gradual decrease in rate. At 8 hours, the edge of the hydrogel dries and The cock is deformed while the middle is still intact and the moisture loss is about 45%. After 12 hours, only the center of the hydrogel remained soft and the moisture loss was approximately 60%. After 18 hours, the water loss was 90%.

Table 2. Three-parallel test to measure the long-term water retention of sample 1 of Example 13.

While the invention has been described with respect to the preferred embodiments the embodiments In addition, many modifications may be made to adapt a particular situation or material to the basic scope without departing from the scope of the invention. Therefore, it is intended that the invention not be limited to the particular embodiments disclosed as the invention

Claims

A sheet-like crosslinked hyaluronate hydrogel comprising:

a) cross-linked hyaluronate hydrogel; and

b) reticular fibers;

Wherein the reticular fibers are embedded in the crosslinked hyaluronate hydrogel and form a cement-reinforced structure.
The sheet-like crosslinked hyaluronate hydrogel according to claim 1, wherein said reticulated fiber is embedded in said crosslinked hyaluronate hydrogel, and a cement-reinforced structure is formed by making it transparent The mixture of the acid salt hydrogel and the reticular fibers is subjected to crosslinking to achieve.
The sheet-like crosslinked hyaluronate hydrogel according to claim 1 or 2, wherein the crosslinked hyaluronate hydrogel is obtained by subjecting a hyaluronate hydrogel to cross-linking.
The sheet-like crosslinked hyaluronate hydrogel according to any one of claims 1 to 3, wherein the reticular fiber is embedded in the crosslinked hyaluronate water by a pressure infusion method or a composite superposition method In the gel.
The sheet-like crosslinked hyaluronate hydrogel according to any one of claims 1 to 4, wherein the crosslinking is selected from the group consisting of physical crosslinking, chemical crosslinking, and self-crosslinking.
A sheet-like crosslinked hyaluronate hydrogel according to claim 5, wherein said physical crosslinking comprises different macromolecules of said hyaluronate under the action of nuclear, optical, electrical, magnetic or ultrasonic energy. Crosslinking occurs between the hydroxyl group, the carbonyl group or the amino group.
The sheet-like crosslinked hyaluronate hydrogel according to claim 5, wherein said chemical crosslinking use comprises having a selected from the group consisting of a diether group, a diepoxy group, a dialdehyde group, a diamino group, a dimercapto group, and a double A crosslinking agent for a compound of an ethylenic group.
The sheet-like crosslinked hyaluronate hydrogel according to claim 7, wherein the chemical crosslinking uses a crosslinking agent comprising a compound having a diepoxy group.
The sheet-like crosslinked hyaluronate hydrogel according to claim 8, wherein said chemical crosslinking uses 1,4 butanediol and glycidyl ether or 1,2,7,8-diepoxyoctane Make As a crosslinking agent.
The sheet-like crosslinked hyaluronate hydrogel according to claim 5, wherein said self-crosslinking is in an ionic medium, and a hydroxyl group, a carbonyl group or an amino group of a different macromolecule of said hyaluronate occurs spontaneously between Crosslinking.
The sheet-like crosslinked hyaluronate hydrogel according to any one of claims 1 to 10, wherein the reticular fibers are composed of a synthetic material, a natural material, or a combination thereof.
The sheet-like crosslinked hyaluronate hydrogel according to claim 11, wherein the synthetic material is a chemical synthetic fiber material selected from the group consisting of polyamide, polypropylene, polyester or polyvinylidene chloride.
The sheet-like crosslinked hyaluronate hydrogel according to claim 11, wherein the natural material is a fibrous material which is grown in nature and is selected from the group consisting of silk, noodles, hemp or hair.
The sheet-like crosslinked hyaluronate hydrogel according to any one of claims 1 to 13, wherein the reticulated fiber is an absorbable reticulated fiber.
The sheet-like crosslinked hyaluronate hydrogel according to any one of claims 1 to 14, wherein the reticular fibers have a pore diameter of from 1 to 30 mm, preferably from 0.5 to 30 mm.
The sheet-like crosslinked hyaluronate hydrogel according to any one of claims 1 to 15, which comprises 7 to 12% by weight, preferably 9 to 10% based on the sheet-like crosslinked hyaluronate hydrogel. % by weight hyaluronate.
The sheet-like crosslinked hyaluronate hydrogel according to any one of claims 1 to 16, which further comprises an additive selected from the group consisting of a bacteriostatic agent, an anesthetic agent, and an epidermal growth factor.
The sheet-like crosslinked hyaluronate hydrogel according to claim 17, which further comprises a bacteriostatic agent.
The sheet-like crosslinked hyaluronate hydrogel according to claim 18, wherein the bacteriostatic agent is cerium nitrate.
The sheet-like crosslinked hyaluronate hydrogel according to any one of claims 1 to 19, which further comprises phosphate-physiological saline.
The sheet-like crosslinked hyaluronate hydrogel according to claim 20, wherein the phosphate-physiological saline is present at 20-80% of a 100% swelling degree.
The sheet-like crosslinked hyaluronate hydrogel according to any one of claims 1 to 21, which has a thickness of from 0.01 to 3 cm, preferably from 0.01 to 2 cm, more preferably from 0.03 to 2 cm.
The sheet-like crosslinked hyaluronate hydrogel according to any one of claims 1 to 2, wherein the hyaluronate is replaced with one selected from the group consisting of chondroitin sulfate, keratan sulfate, and heparin.
A method for preparing a sheet-like crosslinked hyaluronate hydrogel comprising:

a) synthesizing hyaluronic acid water into a hyaluronate hydrogel;

b) pressing the hyaluronate hydrogel onto the reticular fibers into a sheet-like hyaluronate hydrogel in which the reticular fibers are embedded;

c) crosslinking the sheet-like hyaluronate hydrogel.
The method of claim 24 wherein said hyaluronate hydrogel is pressed against opposite sides of said reticulated fibers from opposite directions.
The method according to claim 24 or 25, wherein the crosslinking is carried out under alkaline conditions.
The method according to claim 26, wherein said alkaline condition is a pH of from 9 to 14, preferably a pH of from 13 to 14, more preferably a pH of 13.
The method of claim 26 wherein said crosslinking is carried out in a 1% by weight sodium hydroxide solution.
The method according to any one of claims 24 to 28, wherein the crosslinking uses a group having a group selected from the group consisting of a diether group, a diepoxy group, a dialdehyde group, a bisamino group, a bisindenyl group, and a diene bond. a crosslinking agent for the compound of the group.
The method according to claim 29, wherein the crosslinking uses a compound having a diepoxy group as a crosslinking agent.
The method according to claim 30, wherein said crosslinking uses 1,4 butanediol and glycidyl ether or 1,2,7,8-diepoxyoctane as a crosslinking agent.
The method according to any one of claims 24 to 31, wherein the crosslinking is carried out at a temperature of from 0 to 80 ° C, preferably from 40 to 60 ° C, more preferably 50 ° C.
The method according to any one of claims 24 to 32, wherein the crosslinking The reaction time is at least 1.5 to 3 hours, preferably 2 hours.
A method according to any one of claims 24 to 33, further comprising adjusting the pH of the crosslinked hyaluronate hydrogel to 3-10, preferably 6-8, with an acidic substance.
A dressing comprising the sheet-like crosslinked hyaluronate hydrogel of any one of claims 1 to 23 or a sheet-like cross-linked transparent prepared by the method of any one of claims 24-34 Hydrochloride salt hydrogel.
A mask comprising the sheet-like crosslinked hyaluronate hydrogel of any one of claims 1 to 23 or a sheet-like cross-linked transparent prepared by the method of any one of claims 24-34 Hydrochloride salt hydrogel.
A sheet-like crosslinked hyaluronate hydrogel prepared according to any one of claims 1 to 23 or prepared by the method according to any one of claims 24 to 34 as a dressing, a mask, a sustained release carrier, or Use of a barrier material for adhesion between organ tissues after surgery.
Use of a sheet-like crosslinked hyaluronate hydrogel prepared according to any one of claims 1 to 23 or prepared by the method of any one of claims 24-34 for skin wounds.
A sheet-like crosslinked hyaluronate hydrogel prepared according to any one of claims 1 to 23 or prepared by the method of any one of claims 24-34 for use in burns, ulcers, or laser-clearing of the face Use after skin dark spots.
Use of a sheet-like crosslinked hyaluronate hydrogel prepared according to any one of claims 1 to 23 or prepared by the method of any one of claims 24-34 in the preparation of a stem cell culture scaffold.