Hemostatic aerogel and preparation method thereof

Abstract

The invention relates to the field of medicines, in particular to hemostatic aerogel and a preparation method thereof. The aerogel is prepared from oyster shell-based microspheres and a graft of agarose and gelatin; the oyster shell-based microspheres are dispersed in a graft of agarose and gelatin; the aerogel does not contain water; wherein the oyster shell-based microspheres comprise precipitates obtained by reaction of oyster shell substances and urea. The oyster shell-based microspheres are dispersed in the graft of agarose and gelatin, and do not contain water, so that the aerogel with light weight and high porosity can be obtained; the aerogel is more beneficial to hemostasis. The oyster shell-based microspheres are dispersed in a graft of agarose and gelatin; the oyster shell-based microspheres can be embedded into a network of a graft of agarose and gelatin, so that the porosity of the aerogel is higher, and hemostasis is more facilitated. The excellent porosity is more beneficial to hemostasis; the method is especially suitable for some scenes with a large amount of bleeding.

Description

Technical Field

[0001] The present application relates to the field of medicine, and in particular, to a hemostatic aerogel and a preparation method thereof. Background Art

[0002] Hemostatic aerogel has the function of coagulating wounds. Its advantages of light weight and high porosity enable aerogel to quickly absorb blood, thereby having a certain sealing effect on bleeding wounds, which is better than ordinary bandages.

[0003] However, the current application range of hemostatic aerogels is limited. Summary of the invention

[0004] The purpose of the embodiments of the present application is to provide a hemostatic aerogel and a preparation method thereof.

[0005] In a first aspect, the present application provides a hemostatic aerogel, the aerogel comprising:

[0006] Oyster shell-based microspheres, agarose and gelatin grafts; oyster shell-based microspheres are dispersed in the agarose and gelatin grafts; the aerogel does not contain water;

[0007] The oyster shell-based microspheres include: precipitates of the reaction of oyster shell materials and urea.

[0008] In the above technical solution, oyster shell-based microspheres are dispersed in the graft aerogel of agarose and gelatin, and are water-free, so that a lightweight, high-porosity aerogel can be obtained; this aerogel is more conducive to hemostasis. Oyster shell-based microspheres are dispersed in the graft of agarose and gelatin; the oyster shell-based microspheres can be embedded in the aerogel network of the graft of agarose and gelatin, making the aerogel have a higher porosity and more conducive to hemostasis. Furthermore, the excellent porosity is more conducive to hemostasis; especially suitable for some scenes with heavy bleeding.

[0009] Furthermore, the grafted product of agarose and gelatin forms a composite aerogel with oyster shell-based microspheres, which makes the mechanical properties of the aerogel better, especially the compression performance better, which is more conducive to hemostasis; further, the excellent compression performance can also greatly expand the application range of the aerogel; for some irregular or narrow areas, it can also stop bleeding well.

[0010] In other embodiments of the present application, in terms of mass percentage, the proportion of oyster shell white-based microspheres in the aerogel is 1% to 20%.

[0011] In other embodiments of the present application, the density of the aerogel is 20 mg / cm 3 ~60mg / cm 3 .

[0012] In other embodiments of the present application, the porosity of the aerogel is 70% to 92%.

[0013] In a second aspect, the present application provides a method for preparing a hemostatic aerogel, comprising:

[0014] Oyster shell-based microspheres were dispersed in a graft of agarose and gelatin, and the water was removed by freeze-drying process;

[0015] Wherein, the preparation of the grafted product of agarose and gelatin comprises:

[0016] The agarose solution and gelatin solution were activated separately;

[0017] The activated agarose solution and the activated gelatin solution are mixed and reacted at 50°C to 70°C for 3 to 12 hours; water is then added to obtain a reaction solution; the reaction solution is freeze-dried to obtain a graft of agarose and gelatin;

[0018] The preparation of oyster shell-based microspheres comprises: reacting oyster shell materials with urea to obtain a precipitate.

[0019] In other embodiments of the present application, the agarose solution and the gelatin solution are activated separately, comprising:

[0020] N,N'-Carbonyldiimidazole was added to the agarose solution and the gelatin solution, respectively.

[0021] In other embodiments of the present application, preparing an agarose solution and a gelatin solution comprises:

[0022] Agarose or gelatin was dissolved in anhydrous dimethyl sulfoxide.

[0023] In other embodiments of the present application, oyster shell-based microspheres are dispersed in a graft of agarose and gelatin, and the water is removed by freeze drying, comprising:

[0024] The freeze-dried graft of agarose and gelatin is dissolved to obtain a solution, and oyster shell-based microspheres are added to the solution of the graft of agarose and gelatin to disperse evenly to obtain a mixed solution; and the mixed solution is freeze-dried.

[0025] In other embodiments of the present application, oyster shell material is reacted with urea to obtain a precipitate, comprising:

[0026] The oyster shell material is mixed with citrate and urea to undergo a hydration reaction to obtain a precipitate;

[0027] Optionally, preparing oyster shell material comprises:

[0028] The oyster shells are calcined to obtain a sintered product. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] In order to more clearly illustrate the technical solutions of the embodiments of the present application, the drawings required for use in the embodiments will be briefly introduced below. It should be understood that the following drawings only show certain embodiments of the present application and therefore should not be regarded as limiting the scope. For ordinary technicians in this field, other related drawings can be obtained based on these drawings without paying creative work.

[0030] Figure 1 The pore structure diagram of the aerogels prepared in the comparative example and the embodiment is detected by scanning electron microscopy. DETAILED DESCRIPTION

[0031] In order to make the purpose, technical solutions and advantages of the embodiments of the present application clearer, the technical solutions in the embodiments of the present application will be clearly and completely described below. Obviously, the described embodiments are only part of the embodiments of the present application, not all of the embodiments.

[0032] Therefore, the following detailed description of the embodiments of the present application is not intended to limit the scope of the present application for protection, but merely represents selected embodiments of the present application. Based on the embodiments in the present application, all other embodiments obtained by ordinary technicians in the field without creative work are within the scope of protection of the present application.

[0033] The present application provides a hemostatic aerogel, the aerogel comprising:

[0034] Oyster shell-based microspheres, agarose and gelatin grafts; oyster shell-based microspheres are dispersed in the agarose and gelatin grafts; the aerogel does not contain water;

[0035] The oyster shell-based microspheres include: precipitates of the reaction of oyster shell materials and urea.

[0036] In the above technical solution, oyster shell-based microspheres are dispersed in the graft of agarose and gelatin and do not contain water, so that a lightweight, high-porosity aerogel can be obtained; this aerogel is more conducive to hemostasis. Oyster shell-based microspheres are dispersed in the graft of agarose and gelatin; the oyster shell-based microspheres can be embedded in the network of the graft of agarose and gelatin, making the porosity of the aerogel higher and more conducive to hemostasis. Furthermore, the excellent porosity is more conducive to hemostasis; especially suitable for some scenes with heavy bleeding.

[0037] Furthermore, the grafted product of agarose and gelatin forms a composite aerogel with oyster shell-based microspheres, which makes the mechanical properties of the aerogel better, especially the compression performance better, which is more conducive to hemostasis; further, the excellent compression performance can also greatly expand the application range of the aerogel; for some irregular or narrow areas, it can also stop bleeding well.

[0038] Furthermore, in some embodiments of the present application, in the aerogel, the proportion of oyster shell-based microspheres is 1% to 20% by mass.

[0039] In the above technical solution, the proportion of oyster shell-based microspheres in the aerogel is 1% to 20% by mass.

[0040] In the above technical scheme, the proportion of oyster shell-based microspheres in the aerogel is 1% to 20%; they can be well dispersed and evenly embedded in the network of agarose and gelatin graft aerogel, and are beneficial to improving the mechanical properties of the aerogel, and can achieve the regulation of the pore size of the aerogel, making the pore size of the aerogel smaller and the porosity higher; it is more conducive to hemostasis; thereby effectively improving the pore size and porosity of the composite aerogel.

[0041] Exemplarily, in some embodiments of the present application, in the aerogel, the proportion of oyster shell-based microspheres is 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20% or a range between any two of the foregoing values, by mass percentage.

[0042] Furthermore, in some embodiments of the present application, the density of the aerogel is 20 mg / cm 3 ~60mg / cm 3 .

[0043] In the above technical solution, the density of the aerogel is 20 mg / cm 3 ~60mg / cm 3 , which is conducive to obtaining lightweight aerogel; this lightweight aerogel is conducive to improving the hemostatic effect of aerogel.

[0044] For example, in some embodiments of the present application, the density of the aerogel is 20 mg / cm 3 , 21mg / cm 3 , 22mg / cm 3 , 23mg / cm 3 , 24mg / cm 3 , 25mg / cm 3 、26mg / cm 3 , 27mg / cm 3 、28mg / cm 3 , 29mg / cm 3 、30mg / cm 3 、32mg / cm 3 、35mg / cm 3 、38mg / cm 3 , 40mg / cm 3, 45mg / cm 3 , 48mg / cm 3 、50mg / cm 3 、55mg / cm 3 、56mg / cm 3 , 57mg / cm 3 、58mg / cm 3 、59mg / cm 3 、60mg / cm 3 Or the range between any two of the above values.

[0045] Furthermore, in some embodiments of the present application, the above-mentioned "density of aerogel" has a meaning well known in the art, which represents the mass of the sample per unit volume.

[0046] The measurement can be performed using instruments and methods known in the art.

[0047] Illustratively, in some embodiments of the present application, the above-mentioned test method for the "density of aerogel" includes: (1) measuring the weight (mg) of a cubic sample using an analytical balance; (2) measuring the length (cm), width (cm) and height (cm) of the corresponding cubic sample using a vernier caliper; (3) calculating the density of the aerogel.

[0048] Furthermore, in some embodiments of the present application, the porosity of the aerogel is 70% to 92%.

[0049] In the above technical solution, the porosity of the aerogel is 70% to 92%, which is conducive to obtaining an aerogel with a better pore structure; the porosity of the aerogel is within the above range, which is conducive to improving the hemostatic effect of the aerogel.

[0050] Exemplarily, in some embodiments of the present application, the porosity of the aerogel is 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92% or the range between any two of the foregoing values.

[0051] Furthermore, in some embodiments of the present application, the above-mentioned "porosity of aerogel" has a well-known meaning in the art, which means: the volume occupied by pores in a unit volume of aerogel.

[0052] The measurement can be performed using instruments and methods known in the art.

[0053] For example, in some embodiments of the present application, the above-mentioned test method for the porosity of aerogel includes: (1) measuring the length, width and height of a cubic sample using a vernier caliper; (2) using an analytical balance to measure the initial weight of the cubic sample; (3) immersing the sample in anhydrous ethanol, weighing it after saturation; (4) calculating the porosity of the aerogel.

[0054] Some embodiments of the present application provide a method for preparing a hemostatic aerogel, comprising:

[0055] Oyster shell-based microspheres were dispersed in a graft of agarose and gelatin, and the water was removed by freeze drying;

[0056] Wherein, the preparation of the grafted product of agarose and gelatin comprises:

[0057] The agarose solution and gelatin solution were activated separately;

[0058] The activated agarose solution and the activated gelatin solution are mixed and reacted at 50°C to 70°C for 3 to 12 hours; water is then added to obtain a reaction solution; and the reaction solution is freeze-dried;

[0059] The preparation of oyster shell-based microspheres comprises: reacting oyster shell materials with urea to obtain a precipitate.

[0060] In the above technical solution, oyster shell-based microspheres are dispersed in the graft of agarose and gelatin and do not contain water, so that a lightweight, high-porosity aerogel can be obtained; this aerogel is more conducive to hemostasis. Oyster shell-based microspheres are dispersed in the graft of agarose and gelatin; the oyster shell-based microspheres can be embedded in the network of the graft of agarose and gelatin, making the porosity of the aerogel higher and more conducive to hemostasis. Furthermore, the excellent porosity is more conducive to hemostasis; especially suitable for some scenes with heavy bleeding.

[0061] In some embodiments of the present application, the preparation of the above-mentioned oyster shell-based microspheres includes:

[0062] The oyster shell material is reacted with urea to obtain a precipitate including:

[0063] The oyster shell material is mixed with phosphoric acid, citrate and urea to carry out a hydration reaction to obtain a precipitate.

[0064] Further optionally, the "oyster shell material" mentioned above in the present application can be selected from various shellfish shells such as oyster shells, scallop shells, clam shells, and mussel shells.

[0065] Various shellfish shells, such as oyster shells, scallop shells, clam shells, and mussel shells, are usually treated as waste; how to achieve their green recycling has always been a problem in coastal areas.

[0066] In the above technical scheme, various shellfish shell wastes such as oyster shells, scallop shells, clam shells, and clam shells are used as raw materials for preparing oyster shell-based microspheres, which effectively solves this problem and realizes the recycling of resources, which is conducive to the promotion and application in industry and achieves the effect of reducing costs.

[0067] Further optionally, in some embodiments of the present application, the above-mentioned preparation of oyster shell material includes:

[0068] The oyster shells are calcined to obtain a sintered product.

[0069] Further optionally, in some embodiments of the present application, the oyster shell is calcined at a temperature of 500°C to 600°C.

[0070] Illustratively, in some embodiments of the present application, the oyster shell is calcined at 500°C, 505°C, 510°C, 515°C, 520°C, 525°C, 530°C, 535°C, 540°C, 545°C, 550°C, 555°C, 560°C, 565°C, 570°C, 575°C, 580°C, 585°C, 590°C, 595°C, 600°C or a temperature in a range between any two of the foregoing values.

[0071] Further optionally, in some embodiments of the present application, the oyster shell is calcined for 1 hour to 5 hours.

[0072] Illustratively, in some embodiments of the present application, the oyster shell is calcined for 1 hour, 1.2 hours, 1.5 hours, 1.8 hours, 2 hours, 2.5 hours, 2.8 hours, 3 hours, 3.5 hours, 3.8 hours, 4 hours, 4.2 hours, 4.5 hours, 4.8 hours, 5 hours or a time range between any two of the foregoing values.

[0073] Furthermore, in some embodiments of the present application, the oyster shells need to be crushed and sieved before being calcined.

[0074] Further optionally, in some embodiments of the present application, before calcining the oyster shell, it is also necessary to crush it and pass it through an 80-100 mesh sieve.

[0075] Illustratively, in some embodiments of the present application, before calcining the oyster shell, it is also necessary to crush it and pass it through a sieve with a mesh number of 80 mesh, 82 mesh, 85 mesh, 88 mesh, 90 mesh, 91 mesh, 92 mesh, 93 mesh, 94 mesh, 95 mesh, 96 mesh, 97 mesh, 98 mesh, 99 mesh, 100 or a range between any two of the foregoing values.

[0076] Illustratively, in some embodiments of the present application, calcining oyster shells to obtain a sintered product includes: washing and drying the oyster shells, crushing them using a grinder, and sieving them with a 100-mesh sieve to obtain oyster shell powder with fine particles; and calcining the oyster shell powder in a muffle furnace at 600° C. for 3 hours.

[0077] Furthermore, in some embodiments of the present application, the oyster shell is calcined, and the obtained sintered material is dissolved for subsequent reactions.

[0078] Further optionally, in some embodiments of the present application, a weak acid is used for dissolution.

[0079] For example, in some embodiments of the present application, acetic acid, formic acid, etc. can be selected for dissolution. After dissolution, the solution is used for subsequent reactions to facilitate the preparation of oyster shell-based microspheres.

[0080] Further optionally, in some embodiments of the present application, a weak acid with a mass concentration of 5% to 20% is used to dissolve the sintered oyster shell. Exemplarily, in some embodiments of the present application, a weak acid with a mass concentration of 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20% or a concentration range between any two of the foregoing values ​​is used to dissolve the sintered oyster shell.

[0081] Further optionally, in some embodiments of the present application, the aforementioned dissolved oyster shell solution is also filtered.

[0082] Further, in some embodiments of the present application, the above-prepared oyster shell material solution is reacted with phosphoric acid, urea, and citrate to obtain a precipitate, including:

[0083] The oyster shell material is mixed with phosphoric acid, citrate and urea to carry out a hydration reaction to obtain a precipitate.

[0084] In the above technical solution, oyster shell material is mixed with phosphoric acid, citrate and urea to undergo hydration reaction to obtain a precipitate, thereby facilitating the acquisition of an aerogel structure with excellent pore structure and more conducive to hemostasis.

[0085] Further optionally, in some embodiments of the present application, the above-mentioned citrate can be selected from at least one of sodium citrate, potassium citrate, zinc citrate, and calcium citrate. Exemplarily, the above-mentioned citrate can be selected from any one of sodium citrate, potassium citrate, zinc citrate, and calcium citrate; or in some embodiments of the present application, the above-mentioned citrate can be selected from a mixture of sodium citrate and potassium citrate; or a mixture of zinc citrate and calcium citrate, etc.; each raw material in the above-mentioned mixture can be mixed in any proportion.

[0086] Further optionally, in some embodiments of the present application, oyster shell material is mixed with phosphoric acid, citrate, and urea to carry out a hydration reaction, which can be carried out according to the following ratio:

[0087] In terms of mass percentage, 5% to 25% of oyster shell material, 2% to 15% of phosphoric acid, 2% to 10% of citrate, and 50% to 90% of urea are mixed to carry out a hydration reaction.

[0088] In the above technical solution, the oyster shell material is mixed with phosphoric acid, citrate and urea within the above range to carry out a hydration reaction.

[0089] For example, in some embodiments of the present application, the oyster shell material is mixed with phosphoric acid, citrate, and urea for hydration reaction, which can be carried out according to the following ratio:

[0090] In terms of mass percentage, 5%, 10%, 15%, 20% or 25% of oyster shell material is mixed with 2%-15% of phosphoric acid, 2%-10% of citrate and 50%-90% of urea to carry out a hydration reaction.

[0091] Further optionally, in some embodiments of the present application, the temperature of the hydration reaction is 50°C to 150°C.

[0092] In the above technical solution, by setting the temperature of the hydration reaction to 50°C to 150°C, oyster shell-based microspheres with good performance can be obtained.

[0093] Illustratively, in some embodiments of the present application, the temperature of the hydration reaction is 50°C, 55°C, 60°C, 65°C, 70°C, 75°C, 80°C, 85°C, 90°C, 95°C, 100°C, 115°C, 120°C, 125°C, 130°C, 135°C, 140°C, 145°C, 150°C or a range between any two of the foregoing values.

[0094] Further optionally, in some embodiments of the present application, after the above-mentioned hydration reaction, the precipitate is taken and washed.

[0095] Further optionally, water and alcohol are used for washing. Exemplarily, deionized water and anhydrous ethanol can be selected for washing.

[0096] Further optionally, the washed precipitate is dried to obtain oyster shell-based microspheres.

[0097] Further optionally, in some embodiments of the present application, the drying temperature can be selected to be 50° C. to 80° C. For example, in some embodiments of the present application, the drying temperature is 50° C., 51° C., 52° C., 53° C., 54° C., 55° C., 56° C., 57° C., 58° C., 59° C., 60° C., 61° C., 62° C., 63° C., 64° C., 65° C., 66° C., 67° C., 68° C., 69° C., 70° C., 72° C., 75° C., 78° C., 80° C. or a range between any two of the foregoing values.

[0098] For example, in some embodiments of the present application, oyster shell-based microspheres can be prepared according to the following method:

[0099] The oyster shells were washed and dried, crushed with a grinder, and sieved with a 100-mesh screen to obtain fine-grained oyster shell powder; the oyster shell powder was calcined in a muffle furnace at 600°C for 3h; then, a certain amount of calcined oyster shell powder was added to a certain amount of 10% acetic acid solution, stirred for a certain time, filtered with a 0.22μm filter membrane, and the filtrate was collected; 50mL of 0.06mol / L phosphoric acid was added to the acetic acid solution of oyster shell powder and stirred evenly, a certain amount of sodium citrate solution and urea were also added, and stirring was continued for a certain time until the solution was uniform; the prepared mixed solution was transferred to a hydration reactor, placed at 90°C for reaction for a certain time, and then washed with deionized water and anhydrous ethanol for 3 times respectively, the white precipitate was collected, and dried at 60°C to obtain oyster shell-based microspheres.

[0100] Furthermore, in some embodiments of the present application, the preparation of the graft of agarose and gelatin includes: reacting the activated agarose solution and the activated gelatin solution separately.

[0101] Further, in some embodiments of the present application,

[0102] The agarose solution and the gelatin solution are activated separately, including:

[0103] N,N'-Carbonyldiimidazole was added to the agarose solution and the gelatin solution, respectively.

[0104] Further optionally, in some embodiments of the present application, the activation time of the agarose solution and the gelatin solution is 45-90 minutes.

[0105] Illustratively, in some embodiments of the present application, the time for activating the agarose solution and the gelatin solution, respectively, is 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90 minutes or the range between any two of the foregoing values.

[0106] Further optionally, in some embodiments of the present application, preparing an agarose solution and a gelatin solution comprises:

[0107] Agarose or gelatin was dissolved in anhydrous dimethyl sulfoxide.

[0108] Further optionally, in some embodiments of the present application, agarose or gelatin is dissolved in anhydrous dimethyl sulfoxide, respectively, comprising:

[0109] Weigh a certain amount of agarose and gelatin, add them into anhydrous dimethyl sulfoxide, and stir at 70°C to 100°C until they are completely dissolved.

[0110] Illustratively, in some embodiments of the present application, a certain amount of agarose and gelatin are weighed and added to anhydrous dimethyl sulfoxide, and stirred at 70°C, 72°C, 75°C, 78°C, 80°C, 82°C, 85°C, 88°C, 90°C, 92°C, 95°C, 98°C, 100°C or a temperature range between any two of the foregoing values ​​until completely dissolved.

[0111] Further, in some embodiments of the present application, the preparation of the graft of agarose and gelatin includes:

[0112] The activated agarose solution and the activated gelatin solution are reacted separately.

[0113] Further optionally, in some embodiments of the present application, the activated agarose solution is mixed with the gelatin solution and reacted at 50°C to 70°C; after 3h to 12h, an appropriate amount of deionized water is added to terminate the reaction, and the obtained solution is placed in a dialysis bag with a molecular weight of 3000 to 5000 and dialyzed for a certain period of time, and freeze-dried to obtain a graft of agarose and gelatin.

[0114] Illustratively, in some embodiments of the present application, the activated agarose solution is mixed with the gelatin solution and reacted at 50°C, 51°C, 52°C, 53°C, 54°C, 55°C, 56°C, 57°C, 58°C, 59°C, 60°C, 61°C, 62°C, 63°C, 65°C, 66°C, 67°C, 68°C, 69°C, 70°C or a temperature range between any two of the foregoing values; 3h, 4h, 5h, 6h, 7h, 8h, 9h, 10h, 11h, 12h or After a time in the range between any two of the aforementioned values, an appropriate amount of deionized water is added to terminate the reaction, and the resulting solution is placed in a dialysis bag with a molecular weight of 3000, 3100, 3200, 3300, 3500, 3600, 3700, 3800, 3900, 4000, 4100, 4200, 4300, 4400, 4500, 4600, 4700, 4800, 4900, 5000 or in the range between any two of the aforementioned values, dialyzed for a certain time, and freeze-dried.

[0115] Illustratively, in some embodiments of the present application, the synthesis of a graft of agarose and gelatin includes: weighing a certain amount of agarose and gelatin, respectively adding them to anhydrous dimethyl sulfoxide, stirring at 70-100°C until completely dissolved; after the agarose is completely dissolved, naturally cooling to room temperature, adding a certain amount of N,N'-carbonyldiimidazole, and activating for a certain time; then mixing the activated agarose solution with the gelatin solution, and reacting at 50-70°C; after 3-12 hours, adding an appropriate amount of deionized water to terminate the reaction, and the obtained solution is placed in a dialysis bag with a molecular weight of 3500 and dialyzed for a certain time, and freeze-dried to obtain a graft of agarose and gelatin.

[0116] Further, in some embodiments of the present application, preparing the hemostatic aerogel comprises:

[0117] The oyster shell-based microspheres prepared above are dispersed in the grafted product of agarose and gelatin prepared above, and the water is removed by freeze drying.

[0118] Furthermore, in some embodiments of the present application, the oyster shell-based microspheres prepared above are dispersed in the graft of agarose and gelatin prepared above, comprising:

[0119] Furthermore, in some embodiments of the present application, a certain amount of the aforementioned graft of agarose and gelatin is weighed and added to deionized water, and stirred at 90-100°C until completely dissolved to obtain a graft solution of a certain concentration of agarose and gelatin, and after cooling to 50-60°C, a certain amount of oyster shell-based microspheres is added and dispersed evenly; the mixed solution is poured into a mold, frozen, and then dried by a freeze dryer to obtain an agarose / gelatin graft-oyster shell microsphere composite aerogel.

[0120] Illustratively, in some embodiments of the present application, a certain amount of the aforementioned agarose and gelatin graft prepared is weighed and added to deionized water, and stirred at a temperature of 90°C, 91°C, 92°C, 93°C, 94°C, 95°C, 96°C, 97°C, 98°C, 99°C, 100°C or a temperature in the range between any two of the aforementioned values ​​until completely dissolved to obtain a certain concentration of agarose and gelatin graft solution, and after cooling to a temperature of 50°C, 51°C, 52°C, 53°C, 54°C, 55°C, 56°C, 57°C, 58°C, 59°C, 60°C or a temperature in the range between any two of the aforementioned values, a certain amount of oyster shell-based microspheres is added and evenly dispersed; the mixed solution is poured into a mold, frozen, and then dried by a freeze dryer to obtain an agarose / gelatin graft-oyster shell microsphere composite aerogel.

[0121] The features and performance of the present application are further described in detail below in conjunction with the embodiments:

[0122] Example 1

[0123] A hemostatic aerogel is provided, which is prepared according to the following steps:

[0124] 1. Synthesis of agarose-gelatin grafts

[0125] A certain amount of agarose and gelatin are weighed and added to anhydrous dimethyl sulfoxide respectively, and stirred at 70-100°C until completely dissolved; after the agarose is completely dissolved, the temperature is naturally cooled to room temperature, and a certain amount of N,N'-carbonyldiimidazole is added to activate for a certain time; then the activated agarose solution is mixed with the gelatin solution, and reacted at 50-70°C; after 3-12 hours, an appropriate amount of deionized water is added to terminate the reaction, and the obtained solution is placed in a dialysis bag with a molecular weight of 3500 and dialyzed for a certain time, and freeze-dried to obtain a graft of agarose and gelatin.

[0126] 2. Preparation of oyster shell-based microspheres:

[0127] The oyster shells were washed and dried, crushed with a grinder, and sieved with a 100-mesh screen to obtain fine-grained oyster shell powder; the oyster shell powder was calcined in a muffle furnace at 600°C for 3h; then, a certain amount of calcined oyster shell powder was added to a certain amount of 10% acetic acid solution, stirred for a certain time, filtered with a 0.22μm filter membrane, and the filtrate was collected; 50mL of 0.06mol / L phosphoric acid was added to the acetic acid solution of oyster shell powder and stirred evenly, a certain amount of sodium citrate solution and urea were also added, and stirring was continued for a certain time until the solution was uniform; the prepared mixed solution was transferred to a hydration reactor, placed at 90°C for reaction for a certain time, and then washed with deionized water and anhydrous ethanol for 3 times respectively, the white precipitate was collected, and dried at 60°C to obtain oyster shell-based microspheres.

[0128] 3. Preparation of agarose / gelatin graft / oyster shell microsphere composite aerogel: Weigh a certain amount of agarose and gelatin graft and add it to deionized water, stir at 95°C until completely dissolved to obtain a certain concentration of agarose and gelatin graft solution, cool to 55°C, add a certain amount of oyster shell-based microspheres, and disperse evenly; pour the mixed solution into a mold, freeze it, and dry it through a freeze dryer to obtain agarose / gelatin graft-oyster shell microsphere composite aerogel. See Table 1 for details.

[0129] Examples 2-10, Comparative Examples 1-3

[0130] The difference from Example 1 is:

[0131] a Theoretical mass ratio of agarose and gelatin in the synthesis of agarose-gelatin grafts; b The percentage of N,N'-carbonyldiimidazole relative to the total mass of agarose and gelatin in the synthesis of agarose-gelatin grafts; c The concentration of agarose-gelatin grafts in aqueous solution in the preparation of aerogels; or d The percentage of oyster shell microsphere mass relative to the mass of agarose-gelatin grafts in the preparation of aerogels. Not the same; see Table 1 for details.

[0132]

Performance test

[0133] 1. The morphology and structure of the aerogels prepared in Examples 1-3 and Comparative Examples 1-3 were detected using a scanning electron microscope.

[0134] Test results such as Figure 1 shown.

[0135] from Figure 1 It can be seen that the sample of comparative example 1 has a significant macroporous structure, while the samples of comparative examples 2 and 3 have smaller pore structures. The sample of the embodiment not only presents a significant strip-shaped macroporous structure, but also a large number of small pore structures can be observed in the macropores, and oyster shell-based microspheres are also embedded in the aerogel pore walls.

[0136] The pore structure of the aerogels prepared in each embodiment is better than the pore structure of the aerogels prepared in each comparative example; thus, it is shown that the hemostatic aerogels in the embodiments of the present application are more conducive to hemostasis.

[0137] Among them: the sample of the embodiment not only has both macroporous structure and microporous structure, but also embeds oyster shell-based microspheres with porous structure, which can not only improve the mechanical properties of aerogel, but also change the adsorption rate of aerogel. Therefore, this structure has certain significance for expanding the application field of aerogel.

[0138] 2. Test the mechanical properties of the above embodiments and comparative examples:

[0139] The mechanical properties of agarose / gelatin graft-oyster shell-based microsphere hemostatic aerogel were evaluated by compression performance test, as follows: cylindrical samples (diameter: ~20 mm, height: ~25 mm) were compressed to 80% of the original height of the sample at a compression rate of 1 mm / min using a universal testing machine, the compression stress-strain curve of the sample was recorded, and the compressive strength of the sample at 80% deformation was calculated.

[0140] The test results are shown in Table 2.

[0141] 3. The density of the above-mentioned embodiments and comparative examples is tested as follows:

[0142] Use a vernier caliper to accurately measure the aerogel sample (~1×1×1cm 3 ) and measured its weight (m) using an analytical balance. Finally, the density (ρ, g / cm 3 ):

[0143]

[0144] Wherein: M (g) is the weight of the aerogel; d (cm), w (cm) and h (cm) represent the length, width and height of the aerogel sample respectively.

[0145] The test results are shown in Table 1.

[0146] 4. The porosity of the above embodiments and comparative examples is tested as follows:

[0147] Aerogel samples (~1×1×1cm 3 ) was measured; then, the aerogel sample was placed in a 15 mL centrifuge tube and 5 mL of anhydrous ethanol was added; finally, the centrifuge tube was ultrasonicated for 1 hour, and the sample was taken out and its weight was measured. The porosity (Porosity, %) of the aerogel was calculated by the following formula:

[0148]

[0149] Among them, M o is the original weight of aerogel, g; M e represents the weight of aerogel after filling with anhydrous ethanol; ρ 无水乙醇 is the density of anhydrous ethanol, g / cm 3 ; d (cm), w (cm) and h (cm) represent the length, width and height of the aerogel sample, respectively.

[0150] The test results are shown in Table 1.

[0151] 5. The adsorption performance test method of agarose / gelatin graft-oyster shell microsphere composite aerogel to deionized water (DW), PBS solution (PBS) and simulated body fluid (SBF) is as follows:

[0152] The absorption performance of aerogels was evaluated by classical gravimetric analysis. First, a known weight of dry aerogel was immersed in a solution (deionized water (DW), PBS (pH 7.4), simulated body fluid (FBS)) and the weight was accurately weighed, recorded as W0. Then, at 37°C, the aerogel was allowed to fully absorb the solution. Finally, the aerogel was taken out, the surface moisture was wiped off, and the weight was weighed, recorded as W1. The adsorption ratio (g / g) of the aerogel was calculated using the following formula:

[0153]

[0154] Wherein, W1 represents the weight of aerogel after liquid adsorption, g; W0 represents the original weight of aerogel, g;

[0155] The test results are shown in Table 2.

[0156] 6. Evaluate the hemostatic performance of the above embodiments and comparative examples:

[0157] a. Whole blood coagulation test

[0158] The coagulation index BCI is a standard for evaluating the hemostatic effect of materials. The lower the BCI value, the better the hemostatic effect.

[0159] The testing method of blood coagulation index BCI includes:

[0160] The size is 1×1×1cm 3 The aerogel was placed in the middle of a culture dish with a diameter of 90 mm, and 100 μL of citrate anticoagulated rabbit whole blood and 20 μL of 0.2 mol / L calcium chloride solution were dripped on the sample and allowed to stand for 5 minutes; then 100 mL of deionized water was added and gently shaken to dissolve the incompletely coagulated blood; 100 μL was drawn into a 96-well plate and the absorbance was measured at a wavelength of 540 nm using an ELISA reader. The coagulation index (BCI, %) was calculated by the following formula: BCI (%) = As / Ac × 100%, where As represents the absorbance value of the sample and Ac represents the absorbance value of the control group.

[0161] The BCI test results are shown in Table 3.

[0162] b. Prothrombin time (PT)

[0163] PT determination is an ideal and commonly used screening test for the extrinsic coagulation system, and can also be used as a quantitative test for coagulation factors in the extrinsic pathway and common pathway. First, centrifuge the citrate anticoagulated rabbit whole blood at 4°C at a rate of 3000rpm for 5 minutes, collect the plasma, and set aside. Weigh 1mg of the sample and place it in a 2mL centrifuge tube, and weigh 3 parallels for each sample. Put the anticoagulated plasma, aerogel sample, and PT reagent in a 37°C water bath and incubate for 3 minutes, then add 100μL plasma and 200μL PT reagent, start timing immediately, observe the coagulation of plasma, and stop timing when the plasma appears lumpy and has no fluidity. The recorded time is the prothrombin time.

[0164] The test results are shown in Table 3.

[0165] c. Activated partial thromboplastin time (APTT)

[0166] APTT assay is a relatively sensitive and commonly used screening test for the intrinsic coagulation system, and can also be used as a quantitative test for coagulation factors of the intrinsic pathway. First, citrate anticoagulated rabbit whole blood is centrifuged at 3000rpm for 5 minutes at 4°C, and plasma is collected for later use. Weigh 2mg of sample and place it in a 2mL centrifuge tube and preheat it at 37°C. Add 100μL of preheated plasma and 100μL APTT reagent and shake evenly; incubate in a 37°C water bath for 5 minutes, add 100μL of 0.025mol / L CaCl2 solution, and immediately start timing to observe the coagulation of plasma. Stop timing when the plasma appears lumpy and has no fluidity. The recorded time is the activated partial thromboplastin time.

[0167] The test results are shown in Table 3.

[0168] d. Hemostatic performance test

[0169] The hemostatic effect of agarose / gelatin graft-oyster shell microsphere composite aerogel on ICR mice was evaluated using a liver hemostasis model.

[0170] The anesthetized mice were fixed on the surgical board. After removing the hair, the abdominal cavity of the mouse was gently cut open with a scalpel to completely expose the liver. The tissue fluid around the liver was completely absorbed by the gauze to reduce experimental interference. The weighed filter paper was placed under the liver, and then a wound of about 5 mm was cut on the liver with a scalpel to induce bleeding. The wound was hemostatically treated with agarose / gelatin graft-oyster shell-based microsphere composite aerogel, and untreated mice were used as the control group. After hemostasis, the hemostasis time was recorded, and the filter paper weight was weighed to calculate the blood loss. The test results are shown in Table 3.

[0171] Table 1 Composition of aerogels prepared in various embodiments and comparative examples and their density and porosity results

[0172]

[0173]

[0174] Notes: a Theoretical mass ratio of agarose and gelatin in the synthesis of agarose / gelatin grafts; b The percentage of N,N'-carbonyldiimidazole relative to the total mass of agarose and gelatin in the synthesis of agarose / gelatin grafts; c The concentration of agarose / gelatin grafts in aqueous solution in the preparation of aerogels; d The percentage of oyster shell microspheres mass relative to the mass of agarose / gelatin grafts in the preparation of aerogels.

[0175] From Table 1 above, we can see that:

[0176] The density of the comparative sample is 15.9±1.9 g / cm 3 ~24.3±1.6g / cm 3 The density of all samples in the examples is 23.7±0.5~53.1±3.5g / cm 3 Except for the sample of Example 1, the density of all other example samples is greater than that of the comparative example sample, mainly due to the introduction of high-density oyster shell-based microspheres and the increased concentration of agarose / gelatin grafts.

[0177] The porosity of the comparative samples is 73.2±9.0% to 79.0±4.3%, and the porosity of the sample of Example 1 is 71.4±5.3%, which is close to the comparative example. The porosity of all other exemplary samples (85.0±3.6% to 91.2±5.7%) is significantly greater than that of the comparative samples.

[0178] This shows that the hemostatic aerogel in the embodiment of the present application has a higher porosity, which is more conducive to hemostasis.

[0179] Table 2 Compression performance and adsorption performance results of aerogels prepared in various embodiments and comparative examples

[0180]

[0181] From Table 2 above, we can see that:

[0182] The compressive strength of comparative example 1 when deformed by 80% is 113.8±36.7KPa~145.6±16.6KPa, and the compressive strength of the embodiment when deformed by 80% can reach 180.7±12.8KPa~1075.9±42.4KPa.

[0183] The mechanical properties of the aerogels prepared in each embodiment are better than those of the aerogels prepared in each comparative example;

[0184] The compressive strength of the aerogel increases significantly with the increase of the concentration of agarose / gelatin grafts and the content of oyster shell microspheres. The compressive strength of the sample in Example 4 is the largest when the deformation is 80%, which can reach 1075.9±42.4 KPa.

[0185] Furthermore, it can be seen from Table 2 that:

[0186] The adsorption rates of the comparative samples for deionized water, PBS solution and simulated body fluids are 25.5±1.6g / g, 21.4±1.0g / g and 19.7±1.4g / g, respectively. The adsorption rates of the example samples for deionized water, PBS solution and simulated body fluids can reach 17.8±1.5g / g to 20.6±0.3g / g, 14.8±0.2g / g to 21.8±1.5g / g and 15.0±0.6g / g to 20.5±1.4g / g, respectively. The adsorption rates of most example samples are not significantly different from those of the comparative samples. The adsorption rates of individual example samples are lower than those of the comparative samples, and the possible reason is that the introduction of oyster shell microspheres occupies part of its volume and causes the density of the aerogel to increase.

[0187] Table 3 BCI, APPT, PT test results of aerogels prepared in various embodiments and comparative examples

[0188]

[0189]

[0190] From Table 3 we can see that:

[0191] The BCI value of the comparative sample is 64.0±4.0%~65.0±3.0%. The BCI values ​​of the samples of Example 5 and Example 6 are close to those of the comparative samples, and the BCI values ​​of the other comparative samples (44.3±6.8%~61.3±4.7%) are significantly lower than those of the comparative samples. From the BCI values ​​in Table 3, it can be found that the BCI value of the aerogel is smaller as the content of agarose / gelatin grafts and oyster shell-based microspheres increases. The BCI value of Example 9 is the smallest, which is 44.3±6.8%.

[0192] Furthermore, it can be seen from Table 3 that:

[0193] The PT value of the control group was 20.3±2.1s; the PT values ​​of all comparative samples were 12.6±0.5s to 14.8±1.4s; and the PT values ​​of all example samples were 11.3±0.3s to 17.9±1.0s.

[0194] As shown in Table 3, relative to the control group, the PT values ​​of all comparative samples and example samples were smaller, and there was no significant difference between the PT values ​​of the comparative samples and the PT values ​​of the example samples. The minimum PT value of the example samples was 11.3±0.3s, which was lower than that of all comparative samples. This shows that, relative to the comparative examples, the changes in the composition of the example samples had no negative effect on the prothrombin pathway.

[0195] Furthermore, it can be seen from Table 3 that:

[0196] The APPT value of the control group was 38.6±1.0s; the APPT values ​​of all comparative examples were 26.8±3.0s to 28.3±3.9s; and the APPT values ​​of all embodiments were 26.3±2.9s to 34.5±2.0s.

[0197] As shown in Table 3, the APPT values ​​of the comparative examples and the example samples were lower than those of the control group. Moreover, there was no significant difference in the APPT values ​​between the comparative examples and the example samples. The minimum APPT value of the example sample was 26.3±2.9s, which was lower than that of all the comparative examples. The reason may be that the active groups -OH and PO4 carried by hydroxyapatite 3- The presence of activated coagulation cascade and shortened APTT time.

[0198] Furthermore, it can be seen from Table 3 that:

[0199] The blood loss and hemostasis time of the control group were 0.558±0.034g and 141.0±5.2s, respectively. Obviously, the blood loss and hemostasis time of the mice treated with the comparative examples and the example samples were significantly reduced relative to the control group. After the comparative example samples were treated, the blood loss and hemostasis time of the mice were 0.436±0.028g~0.485±0.038g and 113.2±8.7s~121.9±7.8s, respectively, and then the blood loss and hemostasis time of the mice treated with the example samples were 0.292±0.010g~0.422±0.010g and 84.6±7.5s~104.8±5.2s, respectively.

[0200] As shown in Table 3, the blood loss and hemostasis time of all the samples in the examples were less than those in the comparative example, the fastest hemostasis time was 84.6±7.2s, and the minimum blood loss was 0.292±0.010g. Therefore, compared with the control group and the comparative example, the hemostasis time and blood loss of mice treated with agarose / gelatin graft-oyster shell microsphere composite aerogel were reduced, indicating that agarose / gelatin graft-oyster shell microsphere composite aerogel has a good hemostatic effect.

[0201] The above description is only the preferred embodiment of the present application and is not intended to limit the present application. For those skilled in the art, the present application may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application shall be included in the protection scope of the present application.

Claims

1. A hemostatic aerogel, characterized in that: The aerogel comprises: Oyster shell-based microspheres, agarose and gelatin grafts; the oyster shell-based microspheres are dispersed in the agarose and gelatin grafts; the aerogel does not contain water; Wherein, the oyster shell-based microspheres include: precipitates of the reaction of oyster shell materials and urea.

2. The hemostatic aerogel according to claim 1, characterized in that In terms of mass percentage, the oyster shell-based microspheres account for 1% to 20% of the aerogel.

3. The hemostatic aerogel according to claim 1, characterized in that The density of the aerogel is 20 mg / cm 3 ~60mg / cm 3 .

4. The hemostatic aerogel according to claim 1, characterized in that The porosity of the aerogel is 70% to 92%.

5. The method for preparing the hemostatic aerogel according to any one of claims 1 to 4, characterized in that: include: Oyster shell-based microspheres were dispersed in a solution of agarose and gelatin grafts, and the water was removed by freeze-drying process; Wherein, the preparation of the graft of agarose and gelatin comprises: The agarose solution and gelatin solution were activated separately; The activated agarose solution and the activated gelatin solution are mixed and reacted at 50°C to 70°C for 3 to 12 hours; water is then added to obtain a reaction solution; the reaction solution is freeze-dried to obtain a graft of agarose and gelatin; The preparation of the oyster shell-based microspheres comprises: reacting oyster shell materials with urea to obtain a precipitate.

6. The method for preparing the hemostatic aerogel according to claim 5, characterized in that: include: The agarose solution and the gelatin solution are activated respectively, comprising: N,N'-carbonyldiimidazole is added to the agarose solution and the gelatin solution, respectively.

7. The method for preparing the hemostatic aerogel according to claim 5, characterized in that: include: The agarose solution and the gelatin solution are prepared, comprising: Agarose or gelatin was dissolved in anhydrous dimethyl sulfoxide.

8. The method for preparing the hemostatic aerogel according to claim 5, characterized in that: include: The method of dispersing oyster shell-based microspheres in a graft of agarose and gelatin and removing water by freeze drying comprises: The freeze-dried graft of agarose and gelatin is dissolved to obtain a solution, and oyster shell-based microspheres are added to the solution of the graft of agarose and gelatin to disperse evenly to obtain a mixed solution; and the mixed solution is freeze-dried.

9. The method for preparing the hemostatic aerogel according to claim 5, characterized in that: include: The step of reacting oyster shell material with urea to obtain a precipitate comprises: The oyster shell material is mixed with citrate and urea to carry out a hydration reaction to obtain a precipitate.

10. The method for preparing hemostatic aerogel according to claim 5, characterized in that: include: The preparation of the oyster shell material comprises: The oyster shells are calcined to obtain a sintered product.

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