CN111632189B

CN111632189B - Injectable hydrogel hemostatic based on marine-derived gelatin, and application method thereof

Info

Publication number: CN111632189B
Application number: CN202010463576.7A
Authority: CN
Inventors: 赵晓丽; 边少荃; 潘浩波
Original assignee: Shenzhen Institute of Advanced Technology of CAS
Current assignee: Shenzhen Institute of Advanced Technology of CAS
Priority date: 2020-05-27
Filing date: 2020-05-27
Publication date: 2021-12-14
Anticipated expiration: 2040-05-27
Also published as: CN111632189A

Abstract

The invention relates to the technical field of hemostats, and particularly provides an injectable hydrogel hemostat based on marine-derived gelatin, and an application method thereof. The injectable hydrogel hemostatic based on the marine gelatin comprises the following components in percentage by weight of 100%; 0.1 to 10 percent of chemically modified marine gelatin; 10 to 20 percent of photo-crosslinking gel factor; 0.1 to 15 percent of tissue adhesion factor; 0.1 to 0.5 percent of photoinitiator; and a dissolved amount of a solvent. The injectable hydrogel hemostatic has the advantages of high speed and good hemostatic effect, no additional wound to peripheral tissues in the hemostatic process, degradable property and wide application in hemostasis of accidental wounds or operation wounds of tissues and visceral organs of human bodies or animal bodies.

Description

Injectable hydrogel hemostatic based on marine-derived gelatin, and application method thereof

Technical Field

The invention belongs to the technical field of hemostats, and particularly relates to an injectable hydrogel hemostat based on marine-derived gelatin, and an application method thereof.

Background

Acute bleeding from accidental wounds and surgical procedures is a major medical problem that is prevalent in the world and can seriously threaten the life and health of the patient. Particularly, human organs such as liver and spleen are the most important, because liver and spleen have abundant vascular tissues, acute bleeding on wound surface is not only common but also troublesome when suffering from trauma or performing related surgical operations. How to find a fast and effective hemostatic treatment has been the best strategy to reduce patient mortality.

For acute bleeding caused by internal organ rupture and bleeding wounds with irregular shapes and deeper narrow wounds, the conventional clinical hemostasis method (such as surgical suture or electrotome hemostasis and the like) or hemostasis material (such as hemostatic gauze or hemostatic sponge and the like) or the conventional clinical hemostasis method cannot penetrate into the wounds to effectively block bleeding points, so that the hemostasis effect is not ideal. At the moment, only when the hemostatic material with excellent performance is injected to the bleeding wound surface and is fully plugged, the bleeding can be effectively stopped, so that the death rate of patients is reduced.

With the development of biomaterial science and in order to better cope with medical acute bleeding, researchers developed a series of injectable hemostatic hydrogels, such as fibrin glue and polymer hydrogel. However, the current injectable hemostatic gels have several problems:

(1) the crosslinking reaction is not controllable. The working mechanism of the injectable hemostatic hydrogel is that two precursor solutions generate a crosslinking reaction to form hydrogel to block a hemostatic point, but the crosslinking reaction is uncontrollable, the gel can block an injector if the operation time is too long, and the precursor solution cannot be gelled in time if the operation time is too short, so that hemostasis failure is caused.

(2) The curing speed is slow. The precursor solution which does not form gel after injection is easy to be washed away by blood, or generates leakage and loss, and can not realize rapid hemostasis.

(3) Poor tissue adhesion and insufficient mechanical properties. This can result in failure to effectively seal the wound after curing, resulting in a failure to stop bleeding.

In view of the above problems, researchers have further studied and developed injectable fast light-curable hydrogel hemostatic based on natural polymer gelatin and hyaluronic acid, which can achieve fast curing, but can undergo swelling behavior in a humid environment in vivo, and the volume increase can cause mechanical property reduction and also can cause wound closure failure, and at the same time, the volume increase can press surrounding tissues to cause additional injury.

Disclosure of Invention

The invention provides an injectable hydrogel hemostatic based on marine gelatin, and an application method thereof, and aims to solve at least one problem of mechanical property reduction, sealing failure, additional damage to surrounding tissues and the like caused by swelling behavior of the conventional injectable rapid photo-curing hydrogel hemostatic.

The invention is realized by the following steps:

the injectable hydrogel hemostatic based on the marine gelatin comprises the following components in percentage by weight of 100%;

correspondingly, the injectable hydrogel hemostatic based on the marine gelatin is applied to hemostasis of accidental wounds or operation wounds of tissues and organs of a human body or an animal body.

Accordingly, the method of application of the injectable hydrogel haemostat based on marine-derived gelatin as described above, comprises the steps of: injecting the injectable hydrogel hemostatic agent based on the marine gelatin into a part to be stopped bleeding, and irradiating by using ultraviolet light to solidify the injectable hydrogel hemostatic agent.

The invention has the following beneficial effects:

compared with the prior art, the injectable hydrogel hemostatic based on the marine gelatin provided by the invention realizes injectable photocuring due to the inclusion of chemically modified marine gelatin, a photocrosslinking gel factor, a tissue adhesion factor and a photoinitiator, can achieve a curing effect within 20s, shows strong tissue adhesion and mechanical properties, and has extremely small curing swelling deformation, so that a bleeding wound surface can be rapidly and durably sealed, and the effects of rapid hemostasis for acute wound bleeding and no additional wound on peripheral tissues are finally realized; in addition, the material has degradable property, can be gradually degraded along with the healing of the wound, and has good biological safety.

The injectable hydrogel hemostatic based on the marine gelatin has the effects of rapid hemostasis and no additional trauma to surrounding tissues, so that the hemostatic can be applied to hemostasis of accidental wounds or operation wounds of human or animal body tissues and visceral organs.

The application method of the injectable hydrogel hemostatic based on the marine gelatin provided by the invention has the characteristics of simple operation, short curing time, high hemostasis speed and the like.

Drawings

In order to more clearly illustrate the technical solution in the embodiments of the present invention, the drawings needed to be used in the embodiments will be briefly described below. It is obvious that the drawings in the following description are only some embodiments of the invention, and that for a person skilled in the art, other drawings can be derived from them without inventive effort.

FIG. 1 is a nuclear magnetic resonance (NMR-400MHz) spectrum of methacrylic anhydride modified marine source fish skin gelatin provided in example 1 of the present invention;

FIG. 2 is a Pluronic F127-diacrylate nuclear magnetic resonance (NMR-400MHz) spectrum provided in example 1 of the present invention;

FIG. 3 is a nuclear magnetic resonance (NMR-400MHz) spectrum of Pluronic F127-dibenzoaldehyde formate provided in example 1 of the present invention;

fig. 4 is a photograph showing swelling of an injectable hydrogel hemostatic agent based on marine-derived gelatin prepared according to example 1 of the present invention;

FIG. 5 is a comparison between the hydrogel hemostatic agent prepared by example 1 of the present invention and the hydrogel provided by comparative example 1 before and after swelling;

fig. 6 is a picture of the change before and after swelling of an injectable hydrogel hemostatic agent based on marine-derived gelatin prepared in example 2 of the present invention;

FIG. 7 is a graphical representation of the percent change in volume after swelling of an injectable hydrogel hemostatic agent based on marine-derived gelatin prepared according to example 1 of the present invention versus the hydrogel provided in comparative example 1;

FIG. 8 is a graphical representation of the percent change in mass after swelling of an injectable hydrogel hemostatic agent based on marine-derived gelatin prepared according to example 1 of the present invention versus the hydrogel provided in comparative example 1;

fig. 9 is a graph of the diameter and mass change during swelling of an injectable hydrogel hemostatic agent based on marine-derived gelatin prepared in example 2 of the present invention;

FIG. 10 is a comparison of the liver hemostasis experiments of example 1 and a blank control mouse.

Detailed Description

In order to make the technical problems, technical solutions and advantageous effects to be solved by the present invention more clearly apparent, the present invention is further described in detail below with reference to the following embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.

The invention provides an injectable hydrogel hemostatic based on marine gelatin, which is in a liquid state before use, and can realize a hemostatic effect only by injecting the hemostatic to a corresponding part to be hemostatic and irradiating the hemostatic for 2-20 s by adopting ultraviolet light.

Specifically, the injectable hydrogel hemostatic based on the marine gelatin comprises the following components in percentage by weight of 100 percent:

the marine gelatin used in the chemically modified marine gelatin is derived from marine fish skin gelatin or marine fish scale gelatin, and the used modifier is methacrylic anhydride. The methacrylic anhydride modified marine gelatin (namely the methacrylic anhydride modified marine fish skin gelatin or the methacrylic anhydride modified marine fish scale gelatin) is obtained through the modification of the methacrylic anhydride.

In some embodiments, chemically modified marine-derived gelatin may be obtained by:

dissolving marine gelatin in deionized water to obtain a solution, then dropwise adding a certain amount of methacrylic anhydride, and adjusting the pH value with an alkaline solution to make the pH value between 8 and 9;

and after the reaction is finished, precipitating with ethanol, centrifugally collecting the precipitate, dialyzing, and finally freeze-drying to obtain the chemically modified marine gelatin.

Preferably, the photocrosslinking gelator is a diacrylate-modified poloxamer. The poloxamer of the present invention is a polyoxyethylene-polyoxypropylene-polyoxyethylene (PEO-PPO-PEO) triblock copolymer (hereinafter referred to as Pluronics), and any Pluronics having hydrophilicity is applicable, such as Pluronics F127, Pluronics F68, Pluronics F87, and Pluronics F108. Further preferably, the diacrylate-modified poloxamer is Pluronics F127-diacrylate (PF 127-DA for short).

In some embodiments, PF127-DA may be obtained by:

pluronics F127 was dissolved in anhydrous dichloromethane and a certain amount of triethylamine was added;

dropwise adding a certain amount of acryloyl chloride under the ice-water bath condition for reaction, after the reaction is finished, precipitating with diethyl ether, centrifuging, collecting the precipitate, dialyzing with water, and finally freeze-drying to obtain PF 127-DA.

Preferably, the tissue adhesion factor according to the present invention is at least one selected from the group consisting of compounds having the ability to react with amino groups and can be integrated into a hydrogel network through a photocuring reaction.

Specifically, the compound having the ability to react with an amino group and can be integrated into the hydrogel network through a photocuring reaction is at least one of a dibenzoate-modified poloxamer, an N-succinimidyl acrylate, 4-vinylbenzaldehyde, 3-methacrylamidodopamine, and other compounds having the ability to react with an amino group and can be integrated into the hydrogel network through a photocuring reaction.

Further preferably, the poloxamer modified by the dibenzoate is Pluronics F127-dibenzoate (PF 127-DF).

When a compound containing a catechol group (e.g., 3-methacrylamidodopamine) is used as the tissue adhesion factor, the content thereof in the injectable hydrogel hemostatic agent based on marine-derived gelatin does not exceed 0.5% by weight, otherwise the solidifying effect is affected.

In some embodiments, PF127-DF may be obtained by:

dissolving Pluronics F127 in anhydrous dichloromethane, adding a certain amount of 4-Dimethylaminopyridine (DMAP) and p-aldehyde benzoic acid, and adding a certain amount of Dicyclohexylcarbodiimide (DCC);

after the reaction is finished, precipitating with diethyl ether, centrifuging, collecting precipitate, dialyzing with deionized water, and finally freeze-drying to obtain PF 127-DF.

Further preferably, in the injectable hydrogel hemostatic based on the marine gelatin, the total content of the poloxamer compound is not more than 20%, for example, when only the diacrylate modified poloxamer is contained, the content of the diacrylate modified poloxamer is not more than 20%; when containing both the diacrylate-modified poloxamer and the dibenzoate-modified poloxamer, the total content of the diacrylate-modified poloxamer and the dibenzoate-modified poloxamer is not more than 20%. If the total content of poloxamer compounds exceeds 20%, the injectable hydrogel hemostatic based on marine gelatin loses liquid fluidity at room temperature, becomes a pasty substance, and cannot be used in an injectable manner. When the total content of the poloxamer compounds exceeds 20 percent, the liquid fluidity can be kept only at 10 ℃, so that the complex negative influence is caused to the use.

In the invention, the photoinitiator used should be capable of curing the injectable hydrogel hemostatic agent based on the marine gelatin under ultraviolet irradiation for (2-20) seconds, and if the photoinitiator used is too long in acting time and the curing time is more than 20 seconds, the hemostatic effect is difficult to exert.

Further preferably, the photoinitiator is phenyl (2,4, 6-trimethylbenzoyl) lithium phosphate (abbreviated as LAP), and the initiator can solidify the injectable hydrogel hemostatic based on the marine gelatin within about 2 seconds, so as to achieve rapid and effective hemostasis.

The solvent used in the present invention is not particularly limited as long as it can dissolve chemically modified marine gelatin, photocrosslinking gelator, tissue adhesion factor and photoinitiator, and does not react before light irradiation, and can be prepared in accordance with the concentration required for actual hemostasis.

Preferably, the solvent may be any one of deionized water, phosphate buffer (PBS buffer for short), and physiological saline.

The injectable hydrogel hemostatic based on the marine gelatin can be prepared by the following method:

dissolving chemically modified marine gelatin, photo-crosslinking gelator and tissue adhesion factor in a solvent according to a target ratio to completely dissolve all components to obtain a solution, adding a photoinitiator, uniformly mixing, and storing under ultraviolet illumination for later use.

Because the injectable hydrogel hemostatic based on the marine gelatin is chemically modified, the marine gelatin has better biological activity and can better promote wound healing; the injectable hydrogel hemostatic based on the marine gelatin has extremely low swelling deformation, can keep enough mechanical property and tissue adhesion for a long time, keeps the wound sealing effect for a long time, does not press surrounding tissues, can be gradually degraded along with the healing of a wound due to the degradable characteristic, has no toxic or harmful effect on organisms due to degradable organisms, and has better biological safety. Therefore, the hemostatic bag can be widely applied to hemostasis of accidental wounds of tissues and organs of human or animal bodies or operation wounds.

The injectable hydrogel hemostatic based on the marine gelatin is applicable to any organ of liver, spleen, kidney, intestine, stomach and lung in the hemostasis process.

Specifically, the operation can be performed as follows:

and injecting the injectable hydrogel hemostatic agent to a part to be hemostatic, and irradiating by using ultraviolet light to solidify the injectable hydrogel hemostatic agent.

When the ultraviolet light is irradiated, the wavelength of the ultraviolet light is not more than 405nm, and the longer the wavelength is, the longer the curing time is, the more adverse to rapid hemostasis, and if the wavelength of the ultraviolet light is shorter, the more easily the injury is generated to the surrounding tissues. The preferred wavelength of the UV light is 300nm to 380nm, for example, 365nm UV light can be used.

In order to better explain the technical scheme of the invention, a plurality of specific embodiments are combined for explanation.

Example 1

A preparation method of an injectable hydrogel hemostatic based on marine-derived gelatin comprises the following steps:

(1) synthesis of chemically modified marine gelatin:

dissolving 5.0g of marine fish skin gelatin in 200mL of deionized water, and heating to dissolve at 60 ℃;

cooling to 50 ℃, dropwise adding 1.0mL of methacrylic anhydride, and adjusting the pH value to about 8.6 by using 2mol/L sodium hydroxide solution; stirring and reacting for 2 h;

and after the reaction is finished, adding the reaction solution into 1000mL of absolute ethyl alcohol for precipitation, centrifuging, collecting the precipitate, dissolving the precipitate with deionized water, dialyzing the solution in the deionized water for 3 days, freeze-drying to obtain the methacrylic anhydride modified marine source fish skin gelatin, and collecting for later use.

(2) Synthesis of PF 127-DA:

dissolving 12.6g of Pluronics F127 in 100mL of anhydrous dichloromethane, adding 1.2mL of triethylamine, introducing nitrogen for 30min, cooling to 0 ℃ in an ice-water bath to obtain a Pluronics F127 solution;

adding 650 mu L of acryloyl chloride into 10mL of anhydrous dichloromethane, dropwise adding the mixture into Pluronics F127 solution, and reacting under the protection of nitrogen;

precipitating with glacial ethyl ether, centrifuging, collecting precipitate, and vacuum drying to obtain crude product;

finally, the crude product is dissolved by deionized water, dialyzed in deionized water at 4 ℃ in a dark place for 3 days, frozen and dried, and collected for later use.

(3) Synthesis of PF 127-DF:

dissolving 12.6g of Pluronics F127 in 100mL of anhydrous dichloromethane, adding 900mg of p-aldehyde benzoic acid and 500mg of 4-dimethylaminopyridine, adding 1.24g of dicyclohexylcarbodiimide, and stirring at room temperature to react completely;

filtering the reaction product, removing precipitate, dripping the filtrate into 500mL of ethyl glacial ether for precipitation, centrifugally collecting the precipitate, and performing vacuum drying to obtain a crude product;

dissolving the crude product with dichloromethane for the second time, precipitating in ethyl ether, centrifuging, collecting precipitate, vacuum drying, and repeating for 2 times; and dissolving the product with deionized water, freeze-drying, and collecting for later use.

(4) Respectively carrying out nuclear magnetic resonance (NMR-400MHz) spectrum characterization on the products obtained in the steps (1) to (3), and obtaining results shown in figures 1-3.

FIG. 1 is a nuclear magnetic resonance spectrum of chemically modified marine source fish skin gelatin.

From FIG. 1, two characteristic peaks (marked with asterisks) with chemical shift values around 5.5 can be seen, corresponding to the asterisks marked in the structural formula of the compound in the figure, indicating the success of the chemical modification.

FIG. 2 is a nuclear magnetic resonance spectrum of PF 127-DA.

From fig. 2, it can be seen that the characteristic peak (number 1) with the chemical shift value around 1.0 and the three characteristic peaks (number 2) around 6.0 correspond to the number 1 and the number 2 respectively identified in the compound structural formula in fig. 2, which indicates the successful preparation of the PF127-DA material.

FIG. 3 is a nuclear magnetic resonance spectrum of PF 127-DF.

From fig. 3, it can be seen that the characteristic peak (number 1) with the chemical shift value around 1.0, three characteristic peaks (number 2) around 8.0, and two characteristic peaks (number 3) around 10.0 correspond to the number 1, the number 2, and the number 3 respectively identified in the compound structural formula in fig. 3, indicating that the PF127-DF material is successfully prepared.

(5) Preparation of injectable hydrogel haemostats based on marine-derived gelatin:

adding 10mg of methacrylic anhydride modified marine fish skin gelatin obtained in the step (1), 100mg of PF127-DA obtained in the step (2), 50mg of PF127-DF obtained in the step (3) and 2.5mg of phenyl (2,4, 6-trimethylbenzoyl) lithium phosphate into 740 mu L of PBS buffer (pH 7.4), shaking by vortex until the mixture is fully dissolved, and preserving by avoiding ultraviolet light.

Example 2

(1) the synthesis of chemically modified marine gelatin, PF127-DA, and PF127-DF was the same as that of example 1, except that the marine gelatin was derived from marine fish scale gelatin.

(2) Preparation of injectable hydrogel haemostats based on marine-derived gelatin:

100mg of methacrylic anhydride modified marine fish phosphogelatin, 100mg of PF127-DA, 50mg of PF127-DF and 2.5mg of phenyl (2,4, 6-trimethylbenzoyl) lithium phosphate are added into 750 muL of PBS buffer (pH 7.4), vortexed to fully dissolve the mixture, and the mixture is preserved by avoiding ultraviolet light.

Example 3

(1) the synthesis of chemically modified marine gelatin, PF127-DA and PF127-DF was the same as that of example 1.

adding 100mg of methacrylic anhydride modified marine source fish skin gelatin, 100mg of PF127-DA, 100mg of PF127-DF and 5mg of lithium phenyl (2,4, 6-trimethylbenzoyl) phosphate into 695 mu L of normal saline, and oscillating to be fully dissolved by vortex, and keeping away from ultraviolet light.

Example 4

(1) the synthesis of chemically modified marine gelatin, PF127-DA was the same as in example 1.

10mg of methacrylic anhydride modified marine source fish skin gelatin, 150mg of PF127-DA, 15mg of acrylic acid-N-succinimide ester and 5mg of phenyl (2,4, 6-trimethylbenzoyl) lithium phosphate are added into 820 mu L of deionized water, and the mixture is vortexed until the mixture is fully dissolved, and the mixture is preserved by avoiding ultraviolet light.

Example 5

(1) chemically modified marine gelatin, PF127-DA are the same as in example 1.

adding 50mg of methacrylic anhydride modified marine source fish skin gelatin, 150mg of PF127-DA, 10mg of acrylic acid-N-succinimide ester and 5mg of lithium phenyl (2,4, 6-trimethylbenzoyl) phosphate into 785 mu L of physiological saline, performing vortex oscillation until the materials are fully dissolved, and keeping away from ultraviolet light for storage.

Example 6

adding 50mg of methacrylic anhydride modified marine fish skin gelatin, 150mg of PF127-DA, 1mg of 4-vinylbenzaldehyde and 5mg of lithium phenyl (2,4, 6-trimethylbenzoyl) phosphate into 794 mu L of physiological saline, performing vortex oscillation until the materials are fully dissolved, and keeping the materials under the condition of avoiding ultraviolet light.

Example 7

adding 50mg of methacrylic anhydride modified marine source fish skin gelatin, 150mg of PF127-DA, 5mg of 3-methacrylamide dopamine and 5mg of lithium phenyl (2,4, 6-trimethylbenzoyl) phosphate into 790 mu L of deionized water, and performing vortex oscillation until the materials are fully dissolved, and keeping the materials under the condition of avoiding ultraviolet light.

Comparative example 1

The hydrogel comprises the following chemical components in percentage by weight of 100 percent:

and (3) performance testing:

in order to verify the performances of the injectable hydrogel hemostatic agent based on the marine gelatin obtained in examples 1 to 7 and the hydrogel obtained in comparative example 1, a swelling performance test, a tissue adhesion performance test and an animal hemostasis experiment were performed on the injectable hydrogel hemostatic agent and the hydrogel respectively.

1. Swelling Performance test

(1) A portion of each of the products of examples 1 and 2 and comparative example 1 was irradiated with 365nm UV light for 20 seconds to cure the resulting hydrogel, and the results are shown in FIGS. 4, 5 and 6.

As can be seen from the figures 4-6, the obtained hydrogel is colorless and highly transparent elastic solid, and the colorless and highly transparent hydrogel ensures that the real-time situation of a bleeding point can be conveniently observed in the hemostasis process, thereby being beneficial to timely changing the treatment scheme.

(2) A part amount of the products of example 1 and comparative example 1 was cured using ultraviolet light under the same conditions, respectively, and immersed in PBS buffer, maintained at 37 ℃, and volume and mass changes were observed and measured. It can be seen that the volume of the solution after being soaked in phosphate buffer solution for 12 hours is not changed any more, and the swelling equilibrium is reached. The resulting change in volume and mass is shown in figures 7 and 8.

As can be seen from fig. 7 and 8, the injectable hydrogel hemostatic based on marine-derived gelatin of example 1 had an initial 147% increase in volume and an initial 161% increase in mass. Whereas the hydrogel of comparative example 1 had a volume increase of the initial 300% and a mass increase of the initial 295%.

A portion of the product from example 2 was soaked in PBS buffer and maintained at 37 ℃. Gel diameter and wet weight measurements were made every 12 hours and compared to the initial hydrogel, and the results are shown in figure 9. As can be seen from FIG. 9, the product of example 2 reached swelling equilibrium within 12 hours.

In conclusion, the injectable hydrogel haemostat based on marine-derived gelatin of the present invention has a very low swelling set.

2. Tissue adhesion

The products of examples 1-7 and comparative example 1 were subjected to tissue adhesion using a conventional hydrogel pressure burst test apparatus.

Specifically, 8 hole-shaped wounds with the diameter of 2mm are manufactured on the surface of the circular pigskin, the serial numbers of the wounds are 1# to 8#, then corresponding products are dripped on the surfaces of the wounds 1# to 8# according to the sequence of the embodiments 1 to 7 and the comparative example 1, and the wounds are cured by 365nm ultraviolet radiation for 20 seconds. Then, the pigskin was soaked in phosphate buffer for 12 hours and then taken out. Finally, the injection was pushed to observe the barometer reading when the hydrogel broke, i.e., the adhesive strength of the hydrogel, and the results are shown in Table 1.

Table 1 tissue adhesion performance test

Example one another	Pigskin wound numbering	Barometer reading/mmHg
			Example 1	1#	261
Example 2	2#	262
			Example 3	3#	273
Example 4	4#	326
			Example 5	5#	291
Example 6	6#	276
			Example 7	7#	277
Comparative example 1	8#	101

As can be seen from Table 1, the burst strength of examples 1 to 7 can reach 270mm Hg or more, which is much higher than 120mm Hg of human blood pressure, and is suitable for rapid hemostasis of emergency bleeding.

3. Hemostasis test

Two C57 mice were used as model animals (male, 8-week-old, 25g in weight), and a 1 cm-long defect was made in each mouse liver to simulate an emergency bleeding, and a piece of clean and dry filter paper was placed under the bleeding wound for easy observation.

After 5 seconds of bleeding, hemostasis was performed using the injectable hydrogel hemostatic based on marine-derived gelatin of example 1, with untreated wounds as a blank control. The bleeding of the wound was observed and the results are shown in FIG. 10.

As can be seen from fig. 10, the injectable hydrogel hemostatic prepared from marine gelatin according to example 1 can stop bleeding effectively by dropping the hemostatic on a bleeding site and irradiating the hemostatic with ultraviolet light for 20 seconds, and has a function of rapidly stopping bleeding in acute bleeding.

In conclusion, the injectable hydrogel hemostatic based on the marine gelatin has the advantages of strong tissue adhesion and mechanical property, small curing swelling deformation and quick hemostasis effect. Meanwhile, due to the degradable characteristics of the used marine gelatin and the tissue adhesion factors, the raw materials and the degradation products have no stimulation to organisms, and the biological safety is realized.

The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents and improvements made within the spirit and principle of the present invention are intended to be included within the scope of the present invention.

Claims

1. An injectable hydrogel hemostatic agent based on marine-derived gelatin, which is characterized by comprising the following components in percentage by weight of 100%;

0.1 to 10 percent of chemically modified marine gelatin;

10 to 20 percent of photo-crosslinking gel factor;

0.1 to 15 percent of tissue adhesion factor;

0.1 to 0.5 percent of photoinitiator;

the solvent dissolution amount;

wherein the chemically modified marine gelatin is methacrylic anhydride modified marine gelatin;

the photocrosslinking gelator is poloxamer modified by diacrylate ester;

the tissue adhesion factor is selected from at least one of a dibenzoate-modified poloxamer, an N-succinimidyl acrylate, a 4-vinylbenzaldehyde, a 3-methacrylamidodopamine, or other compounds that are capable of reacting with an amino group and can be incorporated into a hydrogel network through a photocuring reaction.

2. The injectable hydrogel hemostatic agent based on marine-derived gelatin according to claim 1, wherein the methacrylic anhydride-modified marine-derived gelatin is methacrylic anhydride-modified marine-derived fish skin gelatin or fish scale gelatin.

3. The injectable hydrogel hemostatic agent based on marine-derived gelatin according to claim 1 or 2, wherein when the photocrosslinked gelator is a diacrylate-modified poloxamer and the tissue adhesion factor is a dibenzoate-modified poloxamer, the total content of both is not more than 20%.

4. The injectable hydrogel hemostatic agent based on marine-derived gelatin according to claim 1, wherein the photoinitiator is a photoinitiator that cures upon UV irradiation for 2 to 20 seconds.

5. The injectable hydrogel hemostatic agent based on marine-derived gelatin according to claim 1 or 4, wherein the photoinitiator is phenyl (2,4, 6-trimethylbenzoyl) lithium phosphate.

6. The injectable hydrogel hemostatic agent based on marine-derived gelatin according to claim 1, wherein the vehicle is any one of deionized water, phosphate buffer, physiological saline.