CN115703889A - Hydrogel with postoperative hemostasis and anti-adhesion functions and preparation method thereof - Google Patents

Abstract

The invention discloses a hydrogel with postoperative hemostasis and anti-adhesion functions and a preparation method thereof. The main constituent unit of the hydrogel is amino acid, and due to the special cyclization degradation mechanism of The glue can be degraded within 7 days in the body, so it will not cause foreign body reaction and inflammatory reaction in the body, and within 7 days of the high-incidence stage of postoperative adhesion, the anti-fouling properties of polyethylene glycol can be used to form a physical barrier to effectively prevent tissue adhesion And it will degrade automatically after 7 days.

Description

A hydrogel with postoperative hemostasis and anti-adhesion functions and preparation method thereof

technical field

The invention belongs to the field of medical technology, and in particular relates to a hydrogel with postoperative hemostasis and anti-adhesion functions and a preparation method thereof.

Background technique

Postoperative adhesion of organs is a common postoperative complication, and its incidence rate is about 80%. Visceral adhesions can lead to infertility, chronic pain, intestinal obstruction, limited movement and even death, causing a great burden on patients and medical and health services. As a material with biocompatibility, tunable Young's modulus and tissue similarity, hydrogel has been widely developed and applied in life science, clinical medicine, tissue engineering, environmental protection and other fields in recent years. In the field of tissue engineering, hydrogel has been successfully developed as an artificial cornea and has initially achieved small-scale industrial production, which is a great boon for patients with eye diseases and blindness. In addition, in the field of life sciences, hydrogels are also used in soft robots to achieve precise drug delivery. However, in clinical medicine, the more difficult problem faced by surgeons is the problem of postoperative adhesions. Due to the concealment of its onset, it is often difficult for patients to detect it, but its harm cannot be ignored. Due to its high tissue compatibility, hydrogels have great potential to be developed as products or commodities for preventing tissue adhesions. At present, there is no commercial product on the market that specifically solves postoperative adhesion prevention to solve the adhesion problem.

Contents of the invention

The purpose of the present invention is to overcome the above-mentioned shortcoming of prior art, provide a kind of hydrogel and preparation method thereof that have postoperative hemostasis and anti-adhesion functions concurrently, the hydrogel provided has the functions of preventing organ adhesion, organ hemostasis and biodegradable characteristics.

In order to achieve the above object, the present invention adopts the following technical solutions to achieve:

A method for preparing a hydrogel with postoperative hemostasis and anti-adhesion functions, comprising the following steps:

Step 1, dissolving gelatin in a solvent to make a gelatin solution;

Step 2, dissolving polyethylene glycol carboxyl activated ester in a solvent to make a polyethylene glycol carboxyl activated ester solution;

Step 3, after mixing the gelatin solution and the polyethylene glycol carboxyl activated ester solution according to the volume ratio (0.001-1): (0.001-1), a reaction occurs at a reaction temperature of 0-40°C, and a hydrogel is formed after the reaction; The hydrogel contains an amide bond, and the structural formula of the amide bond is VI,

Wherein, n is 10-1000.

A further improvement of the present invention is:

Preferably, in step 1, the gelatin is the first type of gelatin or the second type of gelatin, the first type of gelatin is made by hydrolysis or thermal denaturation of animal collagen tissue, and the second type of gelatin is made by modifying the first type of gelatin become.

Preferably, in step 1, the mass fraction of gelatin in the gelatin solution is 1-25%.

Preferably, in step 2, the structural formula of the polyethylene glycol carboxyl activated ester is I, II, II, IV, wherein, n is 10-1000; specifically,

Preferably, in step 2, in the polyethylene glycol carboxyl activated ester solution, the concentration of polyethylene glycol carboxyl activated is 5-25%.

Preferably, the solvents in step 1 and step 2 are both buffer solutions.

Preferably, the buffer solution is dilute hydrochloric acid buffer, phosphate buffer, phosphate buffered saline, Tris-hydrochloric acid buffer, glycine-hydrochloric acid buffer, disodium hydrogen phosphate-sodium citrate buffer, citric acid- Sodium hydroxide-hydrochloric acid buffer, citric acid-sodium citrate buffer, disodium hydrogen phosphate-potassium dihydrogen phosphate buffer, potassium dihydrogen phosphate-sodium hydroxide buffer, boric acid-borax buffer, borax-hydroxide Sodium buffer or sodium carbonate-sodium bicarbonate buffer.

Preferably, the gelatin concentration in the gelatin solution is 25%, and the polyethylene glycol carboxyl activated ester concentration in the polyethylene glycol carboxyl activated ester solution is 20%.

A hydrogel with postoperative hemostasis and anti-adhesion functions prepared by any of the above-mentioned preparation methods, the hydrogel contains amide bonds of succinyl ester bonds, and the structural formula of the hydrogel is for VI,

Among them, n is 10-1000.

Preferably, the degradation time of the hydrogel in a simulated body fluid environment is 1 hour to 1 month.

Compared with the prior art, the present invention has the following beneficial effects:

The invention discloses a method for preparing a hydrogel with postoperative hemostasis and anti-adhesion functions. In the preparation method, after mixing gelatin solution and polyethylene glycol carboxyl activated ester, aminolysis reaction occurs, and the gelatin can be prepared in a short time. An in-situ crosslinked hydrogel with hemostasis and anti-adhesion was obtained. The hydrogel uses cheap, biodegradable, and easily mass-produced gelatin as a raw material, and utilizes amino-containing gelatin and polyethylene glycol containing succinic acid carboxyl-activated ester to undergo amide reaction. The nucleophilic attack of the reacts the amino group with the ester group to form an amide bond.

The present invention also discloses a hydrogel with postoperative hemostasis and anti-adhesion functions. It degrades within 7 days in the body, so it will not cause foreign body reactions and inflammatory reactions in the body, and within 7 days of the high-incidence stage of postoperative adhesions, the anti-fouling properties of polyethylene glycol can be used to form a physical barrier to effectively prevent tissue adhesions and within 7 days It will automatically degrade after a few days. The gel has a short gelation time and high compressive strength, and can be used in drug sustained-release materials, tissue engineering scaffolds, medical sponges, visceral hemostatic sealants, surface coatings of medical built-in objects, and epidermal hemostatic seal coatings , Coatings for burn treatment, etc. The gelatin can control its degradation time as required, has good biocompatibility, and has the function of hemostasis.

Description of drawings

Fig. 1 is the lap strength performance figure of the hydrogel that embodiment 1 of the present invention makes;

Fig. 2 is the swelling curve figure of the hydrogel that the embodiment of the present invention 2 makes;

Fig. 3 is the degradation curve figure of the hydrogel of embodiment 3 of the present invention;

Fig. 4 is the degradation curve figure of the hydrogel of embodiment 4 of the present invention;

Fig. 5 is the degradation curve figure of the hydrogel of embodiment 5 of the present invention;

Fig. 6 is the degradation curve diagram of the hydrogel of embodiment 6 of the present invention;

Fig. 7 is the degradation curve diagram of the hydrogel of embodiment 7 of the present invention;

Fig. 8 is the effect verification figure of embodiment 8 of the present invention;

(a) The picture is a picture of the rat liver hemorrhage model, (b) The picture is the picture of the effect of the hydrogel on inhibiting the rat liver hemorrhage;

Fig. 9 is the effect verification figure of embodiment 9 of the present invention;

(a) The picture is a picture of rat liver adhesion caused by not using this hydrogel, (b) The picture is a picture of using this hydrogel to inhibit rat liver adhesion;

Fig. 10 is a diagram of relative cell activity of Example 10 of the present invention.

Detailed ways

Below in conjunction with specific preparation method, the present invention is described in further detail:

The postoperative anti-adhesion hydrogel based on gelatin polyethylene glycol carboxyl activated ester provided by the present invention can be quickly formed on the tissue surface in situ after injection of gelatin and polyethylene glycol carboxyl activated ester on the tissue surface through a double-needle syringe. The glue thus plays the functions of hemostasis and anti-visceral adhesion.

In the anti-viscera adhesion hydrogel based on gelatin and polyethylene glycol, the gelatin can be porcine gelatin, fish gelatin, bovine gelatin, etc., all of the first type of gelatin derived from animal collagen tissue hydrolysis and thermal denaturation, and A gelatin of the second type obtained by modification of the gelatin of the first type above. Modification methods include physical modification (salting out, freezing-thawing), blending modification (hyaluronic acid, chitosan, carrageenan, etc., which can form a blend with gelatin or polyethylene glycol, Triethylamine and other low-molecular compounds that can change the glass transition temperature or strength of gelatin) and chemical modification (acrylic acid and its esters modified gelatin, aminated gelatin, etc.), the model can be A type or B type, gel Strength is 1-500.

In the anti-visceral adhesion hydrogel based on gelatin and polyethylene glycol carboxyl activated ester, the polyethylene glycol carboxyl activated ester has structural formulas I, II, III, and IV, wherein n is 10-1000.

The present invention further provides the preparation method of the anti-adhesion hydrogel based on gelatin and polyethylene glycol carboxyl activated ester, comprising the steps of:

(1) prepare described gelatin solution 1, dissolve gelatin in the solvent,

In the above preparation method, in step (1), in the gelatin solution, the mass fraction of the gelatin can be 1% to 25%, preferably 25%, and the solvent can be a pH1-10 buffer solution, including Phosphate Buffer, Phosphate Buffered Saline, Tris-HCl Buffer, Glycine-HCl Buffer, Disodium Hydrogen Phosphate-Sodium Citrate Buffer, Citric Acid-Sodium Hydroxide-HCl Buffer, Citric Acid-Citrate Sodium buffer, disodium hydrogen phosphate-potassium dihydrogen phosphate buffer, potassium dihydrogen phosphate-sodium hydroxide buffer, boric acid-borax buffer, borax-sodium hydroxide buffer, sodium carbonate-sodium bicarbonate buffer.

(2) preparing the polyethylene glycol carboxyl activated ester solution; dissolving the polyethylene glycol carboxyl activated ester in a buffer solution to form a polyethylene glycol carboxyl activated ester solution.

In step (2), in the polyethylene glycol carboxyl activated ester solution, its mass fraction is 5-25%, preferably 20%, and the solvent is a buffer solution with a pH of 1-10, including phosphate buffer , Phosphate buffered saline, Tris-hydrochloric acid buffer, glycine-hydrochloric acid buffer, disodium hydrogen phosphate-sodium citrate buffer, citric acid-sodium hydroxide-hydrochloric acid buffer, citric acid-sodium citrate buffer, Disodium hydrogen phosphate-potassium dihydrogen phosphate buffer, potassium dihydrogen phosphate-sodium hydroxide buffer, boric acid-borax buffer, borax-sodium hydroxide buffer, sodium carbonate-sodium bicarbonate buffer.

(3) The solution 1 and the solution 2 are mixed to undergo an ammonolysis reaction at a reaction temperature of 0-40° C., and an in-situ cross-linked hydrogel with hemostasis and anti-adhesion is obtained within 3 minutes.

Step (3) The volume ratio of the mixed solution 1 to the mixed solution 2 is 0.001:1-1:0.001.

Described gelatin and polyethylene glycol carboxyl activated ester can react to generate the amide bond of succinyl ester bond, and its bond structural formula is as follows:

Among them, n is 10-1000. The n value in the finally generated hydrogel is the same as the n value of the polyethylene glycol carboxy-activated ester.

The degradation cycle of the anti-adhesion hydrogel based on gelatin and PEG in a simulated body fluid environment is 1 hour to 7 days, specifically 1 hour to 1 month. Specifically, it can be 4 days or 6 days. The mechanism is that the cross-linking density of succinimide ester and amino group is different to produce a hydrogel with adjustable degradation time, and the degree of protonation of the amino group of gelatin is different under different pH environments, resulting in the formation of different degrees of cross-linking with carboxyl-activated esters. Hydrogels with different degradation times were obtained. In addition, the degradation time can also be adjusted by adjusting the concentration of gelatin and carboxyl-activated ester while maintaining the same pH. The hydrogel forms a physical barrier in situ in the tissue to prevent postoperative fibroblast infiltration. Maintain the gel state within 3-7 days of the critical period of postoperative adhesions to prevent postoperative adhesions, and then degrade, which will not cause foreign body reactions caused by the gel itself, but will aggravate the adhesions. The anti-adhesion hydrogel based on gelatin and PEG provided by the invention has potential applications in the following fields:

(1) local drug delivery; (2) flexible electronic materials; (3) soft robots; (4) biological patches; (5) bio-inks; (6) wound dressings, hemostasis, wound repair; (7) internal organs (8) nervous system anti-adhesion; (9) skeletal system anti-adhesion.

Embodiment 1,

Take by weighing 1250mg gelatin (pigskin gelatin, type A, bloom 300) and be dissolved in the sodium bicarbonate solution of 3.75mL 0.1M to obtain mixed solution 1, take by weighing 400mg polyethylene glycol carboxyl activated ester (as shown in formula I, wherein , n is 23) was dissolved in 1.6mL of pH=4.0 phosphate buffer solution to obtain mixed solution 2; the pigskin bought from the supermarket was cut into 3cm long and 1cm wide strips and soaked in the PBS buffer solution of pH=7.4 After 20 minutes, preheat in a 37°C incubator for 5 minutes, draw 0.6 mL of mixture 1 and 2 in equal amounts into double-barreled syringes, and inject them evenly on the pigskin of the three groups. After curing for 1 hour, the lap shear strength was measured with a universal tensile machine, as shown in Figure 1, the lap shear strength was 3.57kPa.

Embodiment 2,

Take by weighing 1250mg gelatin and be dissolved in the phosphate solution of 3.75mL PB=9.0 to obtain mixed solution 1, take by weighing 400mg polyethylene glycol carboxyl activated ester (as shown in formula I, wherein, n=23) be dissolved in 1.6mL pH= Mixed solution 2 was obtained in 4.0 phosphate buffer solution; 0.5mL mixed solution 1 and 2 were drawn in equal amounts with a double-barreled syringe, and then injected on the parafilm and waited for gelation, then randomly cut into 4 pieces and placed in the Fill it with 3mL of simulated body fluid, then place it in a shaker at 37°C (rotating at 100rpm), weigh at preset times 12, 24, 36 and 48h, and record the swelling curve of the hydrogel. As shown in Figure 2, after 48 h, the hydrogel swelled to about 4 times its initial weight.

Embodiment 3,

Take by weighing 1250mg gelatin and be dissolved in the phosphate solution of 3.75mL PB=9.0 to obtain mixed solution 1, take by weighing 400mg polyethylene glycol carboxyl activated ester (as shown in formula I, wherein, n=23) be dissolved in 1.6mL pH= Mixed solution 2 was obtained in the PB buffer solution of 4.0; 0.5mL mixed solution 1 and 2 were drawn in equal amounts with a double-barreled syringe, and then injected on the parafilm and waited for gelation, then randomly cut into 4 pieces and placed in the container. 3mL of simulated body fluid was placed in a shaker at 37°C (100rpm), weighed at preset times of 24, 48, 96, 144h and 192h, and the degradation curve of the hydrogel was recorded. As shown in Figure 3, after 192 h, the hydrogel was completely degraded in the simulated body fluid.

Example 4

Take by weighing 1250mg gelatin and be dissolved in the phosphate solution of 3.75mL PB=7.0 to obtain mixed solution 1, take by weighing 400mg polyethylene glycol carboxyl activated ester (as shown in formula I, wherein, n=23) be dissolved in 1.6mL pH= Mixed solution 2 was obtained in the PB buffer solution of 7.0; 0.5mL mixed solution 1 and 2 were drawn in equal amounts with a double-barreled syringe, and then injected on the parafilm and waited for gelation, then randomly cut into 4 pieces and placed in the container. 3mL of simulated body fluid was placed in a shaker at 37°C (100rpm), weighed at preset times of 24, 48, 96 and 144h, and the degradation curve of the hydrogel was recorded. As shown in Figure 4, after 144 h, the hydrogel was completely degraded in simulated body fluid.

Example 5

Take by weighing 1250mg gelatin and be dissolved in the phosphate solution of 3.75mL PB=9.0 to obtain mixed solution 1, take by weighing 400mg polyethylene glycol carboxyl activated ester (as shown in formula I, wherein, n=23) be dissolved in 1.6mL pH= Mixed solution 2 was obtained in the PB buffer solution of 7.0; 0.5mL mixed solution 1 and 2 were drawn in equal amounts with a double-barreled syringe, and then injected on the parafilm and waited for gelation, then randomly cut into 4 pieces and placed in the container. 3mL of simulated body fluid was placed in a shaker at 37°C (100rpm), weighed at preset times of 24, 48, 96, 144h and 192h, and the degradation curve of the hydrogel was recorded. As shown in Figure 5, after 192 h, the hydrogel was completely degraded in the simulated body fluid.

Example 6

Take by weighing 1000mg gelatin and dissolve in the phosphate solution of 4mL PB=9.0 to obtain mixed solution 1, take by weighing 150mg polyethylene glycol carboxyl activated ester (as shown in formula I, wherein, n=23) dissolve in 850mL pH=7.0 Mixed solution 2 was obtained in PB buffer solution; 0.5mL mixed solution 1 and 2 were drawn in equal amounts with a double-barreled syringe, and then injected on the parafilm and waited for gelation, then randomly cut into 4 pieces and placed in a 3mL container They were placed in a simulated body fluid, and then placed in a shaker at 37°C (rotating at 100 rpm), weighed at preset times of 24, 48, 96, and 144 hours, and recorded the degradation curve of the hydrogel. As shown in Figure 6, after 96 h, the hydrogel was completely degraded in the simulated body fluid.

Combining Example 3, Example 4 and Example 5, it can be concluded that under different pH conditions, different protonation of amino groups on gelatin chains leads to different cross-linking densities, resulting in hydrogels with different degradation times.

In addition, through Examples 5 and 6, it can be concluded that different concentrations of carboxyl-activated esters can also produce hydrogels with different degradation times, because the concentration of carboxyl-activated esters determines the formation of different amounts of covalent gels after crosslinking. The bond further determines the degradation time.

Example 7

Weigh 1250mg gelatin and dissolve in 3.75mL PB=9.0 borate solution to obtain mixed solution 1, weigh 400mg polyethylene glycol carboxyl activated ester (as shown in formula I, wherein, n=23) and dissolve in pH=4.0 Mixed solution 2 was obtained in the PB buffer solution; 0.5mL mixed solution 1 and 2 were drawn in equal amounts with a double-barreled syringe, and then injected on the parafilm and waited for gelation, then randomly cut into 4 pieces and placed in a 3mL simulated body fluid, then placed in a shaker at 37°C (100rpm), weighed at preset times 24, 48, 96, 144h and 192h, and recorded the degradation curve of the hydrogel. As shown in Figure 7, after 192 h, the hydrogel was completely degraded in the simulated body fluid.

Example 8

Weigh 1250mg of gelatin and dissolve in 3.75mL of PB=9.0 solution to obtain mixed solution 1, weigh 400mg of polyethylene glycol carboxyl activated ester (as shown in formula I, wherein, n=23) and dissolve in the PB buffer of pH=4.0 Mixed solution 2 was obtained from the mixed solution; 0.5mL mixed solution 1 and 2 were drawn in equal amounts with a double-barreled syringe, and then the hydrogel with a degradation time of about 7 days was evenly spread on the injured SD rat liver (the wound length was 1cm), as shown in Figure 8, can effectively prevent blood from flowing out.

Example 9

Weigh 1250mg of gelatin and dissolve in 3.75mL PB=9.0 PB solution to obtain mixed solution 1, weigh 400mg of polyethylene glycol carboxyl activated ester (as shown in formula I, wherein, n=23) and dissolve in PB of pH=4.0 Mixed solution 2 was obtained in the buffer solution; 0.5mL mixed solution 1 and mixed solution 2 were drawn in equal amounts with a double-barreled syringe, and then the hydrogel with a degradation time of about 7 days optimized by the cross-linking ratio and concentration was evenly coated. Distributed on the liver of injured SD rats (a square with a wound side length of 0.5 cm), as shown in Figure 9, compared with the control group, it can effectively prevent the adhesion between the liver and surrounding tissues after 4 days.

Example 10

Weigh 1250mg gelatin and dissolve it in the PB solution of 3.75mL PB=9.0 to obtain mixed solution 1, weigh 400mg polyethylene glycol carboxyl activated ester (as shown in formula IV, wherein, n=23) and dissolve it in the PB solution of pH=4.0 Mixed solution 2 was obtained in the buffer solution; draw 0.5mL mixed solution 1 and mixed solution 2 with a double-barreled syringe respectively, and then inject mixed solutions 1 and 2 on the parafilm to form a gel, and prepare the same gel by the same method , and respectively placed the two gels in 10mL DMEM-H medium at 37°C to obtain the 24h extract and 192h degradation solution to test their cytotoxicity, as shown in Figure 10, whether it is the extraction solution or the degradation solution solutions were not cytotoxic.

Example 11

Take gelatin and dissolve it in phosphate buffered saline physiological solution to obtain a mixed solution 1 with a mass fraction of 1%, weigh polyethylene glycol carboxyl activated ester (as shown in formula II, n=30) and dissolve it in borax-sodium hydroxide buffer solution 2 with a mass fraction of 25%; use a double-barreled syringe to draw mixed solution 1 and mixed solution 2 respectively, and then inject mixed solutions 1 and 2 on the parafilm according to the volume ratio of 1:0.01. Gel, using the same method to prepare the same gel.

Example 12

Weigh gelatin and dissolve it in Tris-hydrochloric acid buffer to obtain a mixed solution 1 with a mass fraction of 25%, weigh polyethylene glycol carboxyl activated ester (as shown in formula III, n=40) and dissolve it in sodium carbonate-sodium bicarbonate In the buffer solution, the solution 2 with a mass fraction of 5% was obtained; the mixed solution 1 and the mixed solution 2 were sucked up with a double-barreled syringe, and then the mixed solution 1 and 2 were injected on the parafilm according to the volume ratio of 0.01:1, at 35 ° C Under the gel, the same method was used to prepare the same gel.

Example 13

Take gelatin and dissolve it in glycine-hydrochloric acid buffer to obtain a mass fraction of 5% mixed solution 1, weigh polyethylene glycol carboxyl activated ester (as shown in formula IV, n=23) and dissolve it in borax-sodium hydroxide buffer solution 2 with a mass fraction of 10%; use a double-barreled syringe to draw mixed solution 1 and mixed solution 2 respectively, and then inject mixed solutions 1 and 2 on the parafilm according to the volume ratio of 1:0.001. Gel, using the same method to prepare the same gel.

Example 15

Weigh gelatin and dissolve it in disodium hydrogen phosphate-sodium citrate buffer to obtain a mixed solution 1 with a mass fraction of 10%, weigh polyethylene glycol carboxyl activated ester (as shown in formula II, n=300) and dissolve it in phosphoric acid In the potassium dihydrogen potassium-sodium hydroxide buffer solution, the solution 2 with a mass fraction of 15% is obtained; draw the mixed solution 1 and the mixed solution 2 with a double-barreled syringe, and then inject the mixed solution 1 and 2 according to the volume ratio of 0.1:1 in On the parafilm, gel was formed at 25°C, and the same gel was prepared by the same method.

Example 14

Weigh gelatin and dissolve in citric acid-sodium hydroxide-hydrochloric acid buffer solution to obtain a mass fraction of 15% mixed solution 1, weigh polyethylene glycol carboxyl activated ester (as shown in formula II, n=400) and dissolve in phosphoric acid In the disodium hydrogen-potassium dihydrogen phosphate buffer solution, the solution 2 with a mass fraction of 8% was obtained; the mixed solution 1 and the mixed solution 2 were sucked up with a double-barreled syringe, and then the mixed solution 1 and 2 were injected according to the volume ratio of 1:0.1 On parafilm, gel was formed at 20°C, and the same gel was prepared by the same method.

Example 15

Weigh gelatin and dissolve it in citric acid-sodium citrate buffer solution to obtain a mixed solution 1 with a mass fraction of 8%, weigh polyethylene glycol carboxyl activated ester (as shown in formula III, n=100) and dissolve it in citric acid- In the sodium hydroxide-hydrochloric acid buffer solution, the solution 2 with a mass fraction of 12% was obtained; the mixed solution 1 and the mixed solution 2 were sucked up with a double-barreled syringe, and then the mixed solution 1 and 2 were injected into the parafilm according to the volume ratio of 1:0.01 Above, gel was formed at 15°C, and the same gel was prepared by the same method.

Example 16

Weigh gelatin and dissolve in disodium hydrogen phosphate-potassium dihydrogen phosphate buffer to obtain a mixed solution 1 with a mass fraction of 12%, weigh polyethylene glycol carboxyl activated ester (as shown in formula III, n=400) and dissolve in In the citric acid-sodium citrate buffer solution, the solution 2 with a mass fraction of 18% was obtained; the mixed solution 1 and the mixed solution 2 were sucked up with a double-barreled syringe, and then the mixed solution 1 and 2 were injected into the seal according to the volume ratio of 0.5:1 On the membrane, gel was formed at 10°C, and the same gel was prepared by the same method.

Example 17

Weighing gelatin and dissolving it in potassium dihydrogen phosphate-sodium hydroxide buffer to obtain a mixed solution 1 with a mass fraction of 18%, weighing polyethylene glycol carboxyl activated ester (as shown in formula IV, n=240) dissolved in phosphoric acid In the disodium hydrogen-sodium citrate buffer solution, the solution 2 with a mass fraction of 20% was obtained; the mixed solution 1 and the mixed solution 2 were drawn respectively with a double-barreled syringe, and then the mixed solution 1 and 2 were injected into the On the parafilm, gel was formed at 5°C, and the same gel was prepared by the same method.

Example 18

Weigh gelatin and dissolve it in borax-sodium hydroxide buffer to obtain a mixed solution 1 with a mass fraction of 20%, weigh polyethylene glycol carboxyl activated ester (as shown in formula IV, n=48) and dissolve it in glycine-hydrochloric acid buffer solution 2 with a mass fraction of 20%; use a double-barreled syringe to draw mixed solution 1 and mixed solution 2 respectively, and then inject mixed solutions 1 and 2 on the parafilm according to the volume ratio of 0.05:1. Gel, using the same method to prepare the same gel.

Example 19

Weigh gelatin and dissolve it in borax-sodium hydroxide buffer solution to obtain a mixed solution 1 with a mass fraction of 22%, weigh polyethylene glycol carboxyl activated ester (as shown in formula II, n=400) and dissolve it in Tris-hydrochloric acid buffer solution 2 with a mass fraction of 25%; use a double-barreled syringe to draw mixed solution 1 and mixed solution 2 respectively, and then inject mixed solutions 1 and 2 on the parafilm according to the volume ratio of 1:0.05. Gel, using the same method to prepare the same gel.

Example 20

Weigh gelatin and dissolve it in sodium carbonate-sodium bicarbonate buffer to obtain a mixed solution 1 with a mass fraction of 25%, weigh polyethylene glycol carboxyl activated ester (as shown in formula III, n=100) and dissolve it in Tris-hydrochloric acid In the buffer solution, the solution 2 with a mass fraction of 20% was obtained; the mixed solution 1 and the mixed solution 2 were sucked up with a double-barreled syringe, and then the mixed solution 1 and 2 were injected on the parafilm according to the volume ratio of 1:1. Under the gel, the same method was used to prepare the same gel.

Example 21

Weigh gelatin and dissolve it in borax-sodium hydroxide buffer to obtain a mixed solution 1 with a mass fraction of 22%, weigh polyethylene glycol carboxyl activated ester (as shown in formula II, n=10) and dissolve it in Tris-hydrochloric acid buffer solution 2 with a mass fraction of 25%; use a double-barreled syringe to draw mixed solution 1 and mixed solution 2 respectively, and then inject mixed solutions 1 and 2 on the parafilm according to the volume ratio of 1:0.05. Gel, using the same method to prepare the same gel.

Example 22

Weigh gelatin and dissolve it in sodium carbonate-sodium bicarbonate buffer to obtain a mixed solution 1 with a mass fraction of 25%, weigh polyethylene glycol carboxyl activated ester (as shown in formula III, n=1000) and dissolve it in Tris-hydrochloric acid In the buffer solution, the solution 2 with a mass fraction of 20% was obtained; the mixed solution 1 and the mixed solution 2 were sucked up with a double-barreled syringe, and then the mixed solution 1 and 2 were injected on the parafilm according to the volume ratio of 1:1. Under the gel, the same method was used to prepare the same gel.

Example 23

Weigh gelatin and dissolve it in borax-sodium hydroxide buffer to obtain a mixed solution 1 with a mass fraction of 22%, weigh polyethylene glycol carboxyl activated ester (as shown in formula II, n=500) and dissolve it in Tris-hydrochloric acid buffer solution 2 with a mass fraction of 25%; use a double-barreled syringe to draw mixed solution 1 and mixed solution 2 respectively, and then inject mixed solutions 1 and 2 on the parafilm according to the volume ratio of 1:0.05. Gel, using the same method to prepare the same gel.

Example 24

Weigh gelatin and dissolve it in sodium carbonate-sodium bicarbonate buffer to obtain a mixed solution 1 with a mass fraction of 25%, weigh polyethylene glycol carboxyl activated ester (as shown in formula III, n=800) and dissolve it in Tris-hydrochloric acid In the buffer solution, the solution 2 with a mass fraction of 20% was obtained; the mixed solution 1 and the mixed solution 2 were sucked up with a double-barreled syringe, and then the mixed solution 1 and 2 were injected on the parafilm according to the volume ratio of 1:1. Under the gel, the same method was used to prepare the same gel.

The above descriptions are only preferred embodiments of the present invention, and are not intended to limit the present invention. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principles of the present invention shall be included in the scope of the present invention. within the scope of protection.

Claims (10)

1. A preparation method of hydrogel with postoperative hemostasis and anti-adhesion functions is characterized by comprising the following steps:

step 1, dissolving gelatin in a solvent to prepare a gelatin solution;

step 2, dissolving the polyethylene glycol carboxyl activated ester in a solvent to prepare a polyethylene glycol carboxyl activated ester solution;

and 3, mixing the gelatin solution and the polyethylene glycol carboxyl activated ester solution according to the volume ratio (0.001-1): (0.001-1), reacting at the temperature of 0-40 ℃ to generate hydrogel; the hydrogel contains amido bond, the structural formula of the amido bond is VI,

wherein n is 10-1000.

2. The method for preparing a hydrogel having functions of hemostasis after operation and adhesion prevention according to claim 1, wherein in step 1, the gelatin is a first gelatin or a second gelatin, the first gelatin is prepared by hydrolyzing or thermally denaturing animal collagen tissue, and the second gelatin is prepared by modifying the first gelatin.

3. The method for preparing the hydrogel with the functions of postoperative hemostasis and adhesion prevention according to claim 1, wherein in the step 1, the mass fraction of gelatin in the gelatin solution is 1-25%.

4. The method for preparing the hydrogel with the functions of postoperative hemostasis and adhesion prevention according to claim 1, wherein in the step 2, the polyethylene glycol carboxyl activated ester has the structural formula I, II and IV, wherein n is 10-1000; in particular, the method comprises the following steps of,

5. the method for preparing a hydrogel with post-operative hemostasis and anti-adhesion functions as claimed in claim 1, wherein in step 2, the concentration of activated polyethylene glycol carboxyl in the polyethylene glycol carboxyl activated ester solution is 5-25%.

6. The method for preparing hydrogel with functions of postoperative hemostasis and adhesion prevention according to claim 1, wherein the solvents in step 1 and step 2 are buffer solutions.

7. The method of claim 6, wherein the buffer solution is a dilute hydrochloric acid buffer, a phosphate buffered saline, a Tris-hydrochloric acid buffer, a glycine-hydrochloric acid buffer, a disodium hydrogen phosphate-sodium citrate buffer, a citric acid-sodium hydroxide-hydrochloric acid buffer, a citric acid-sodium citrate buffer, a disodium hydrogen phosphate-potassium dihydrogen phosphate buffer, a potassium dihydrogen phosphate-sodium hydroxide buffer, a boric acid-borax buffer, a borax-sodium hydroxide buffer, or a sodium carbonate-sodium bicarbonate buffer.

8. The method for preparing a hydrogel with functions of postoperative hemostasis and adhesion prevention according to claim 1, wherein the concentration of gelatin in the gelatin solution is 25%, and the concentration of the polyethylene glycol carboxyl activated ester in the polyethylene glycol carboxyl activated ester solution is 20%.

9. A hydrogel with postoperative hemostasis and anti-adhesion functions prepared by the preparation method of any one of claims 1-8, wherein the hydrogel contains amido bonds of succinyl ester bonds, the structural formula of the hydrogel is VI,

wherein n is 10 to 1000.

10. The hydrogel with the functions of hemostasis after operation and adhesion prevention as claimed in claim 9, wherein the degradation time of the hydrogel in the environment of simulated body fluid is 1 hour to 1 month.

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