A zeolite-loaded regenerated cellulose aerogel hemostatic material and preparation method thereof

Abstract

The present invention is applicable to the technical field of hemostatic materials, and provides a method for preparing a zeolite-loaded regenerated cellulose aerogel hemostatic material, comprising the following steps: dissolving carboxymethylated cellulose and polyethylene glycol in an alkaline solution to obtain a cellulose regeneration solution, subsequently adding zeolite and stirring evenly to obtain a white, stable zeolite-regenerated cellulose homogenate; placing the zeolite-regenerated cellulose homogenate in an acid / alcohol coagulation bath for fiber regeneration to obtain a zeolite-regenerated cellulose hydrogel; dialyzing the zeolite-regenerated cellulose hydrogel to neutrality and removing salt, and freeze-drying to obtain a zeolite-loaded regenerated cellulose aerogel hemostatic material. An embodiment of the present invention also provides a zeolite-loaded regenerated cellulose aerogel hemostatic material. The hemostatic material prepared by the present invention has the advantages of fast hemostasis speed, mild exothermic reaction, and biosafety.

Description

Technical Field

[0001] The invention belongs to the technical field of hemostatic materials, and in particular relates to a zeolite-loaded regenerated cellulose aerogel hemostatic material and a preparation method thereof. Background Art

[0002] Excessive blood loss is one of the leading causes of death from trauma, and uncontrolled wound infection can also be life-threatening. The first 10 minutes after a traumatic injury are crucial for emergency treatment and improving the success rate of rescue. The body's own hemostasis mechanism is complex and time-consuming. When massive traumatic bleeding occurs, it is difficult to quickly stop bleeding relying solely on the body. External intervention, namely the use of hemostatic materials, is necessary to achieve rapid hemostasis. For an ideal hemostatic material, hemostatic efficacy, biocompatibility, safety, and operability are essential parameters.

[0003] Currently, commonly used hemostatic materials include chitosan, cellulose, sodium alginate, and new polymers. Organic and mineral hemostatic materials include zeolite, kaolin, and silica. Zeolite is a classic hemostatic agent, widely used since its FDA approval in 2002. Zeolite rapidly absorbs water and, upon contact with a wound, rapidly absorbs plasma, enriches blood cells, and other components, thereby rapidly controlling bleeding. Furthermore, its inert composition is non-toxic to living organisms. However, the use of exposed zeolite can cause severe thermal damage and necrosis of surrounding tissue. Prehydrated zeolite hemostatic materials (ACS+) have been developed in the prior art. These materials produce less exothermic reactions and minimize thermal damage, but their hemostatic efficacy is reduced, and wound tissue temperatures can still rise to 50°C. Furthermore, the zeolite is only physically adsorbed to the material, resulting in a weak bond. Zeolite is non-biodegradable and easily disperses within the wound during the hemostatic process, making it difficult to remove and leading to tissue inflammation and thrombosis.

[0004] Therefore, how to reduce the reaction heat of zeolite absorbing water and improve the stability of the combination of zeolite and materials without reducing the hemostatic effect is of great research significance. Summary of the Invention

[0005] The purpose of the embodiments of the present invention is to provide a method for preparing a zeolite-loaded regenerated cellulose aerogel hemostatic material, aiming to solve the problems raised in the above background technology.

[0006] The embodiment of the present invention is achieved by providing a method for preparing a zeolite-loaded regenerated cellulose aerogel hemostatic material, comprising the following steps:

[0007] Dissolve carboxymethylated cellulose and polyethylene glycol in an alkaline solution to obtain a cellulose regeneration solution, then add zeolite and stir evenly to obtain a white stable zeolite-regenerated cellulose slurry;

[0008] Placing the zeolite-cellulose regenerated cellulose homogenate in an acid / alcohol coagulation bath for fiber regeneration to obtain a zeolite-regenerated cellulose hydrogel, wherein the acid / alcohol coagulation bath is a mixture of an acid solution and an alcohol, wherein the acid solution is 10% by mass sulfuric acid and the alcohol is 50% by mass ethanol, and the mass ratio of the acid solution to the alcohol is 8:92;

[0009] The zeolite-regenerated cellulose hydrogel was dialyzed to neutrality and salt was removed, and then freeze-dried to obtain the zeolite-loaded regenerated cellulose aerogel hemostatic material.

[0010] Preferably, the carboxymethyl substitution degree of the carboxymethylated cellulose is 0.10-0.50.

[0011] Preferably, the alkaline solution is sodium hydroxide or potassium hydroxide.

[0012] Preferably, the mass concentration of carboxymethyl cellulose in the cellulose regeneration solution is 3-10%.

[0013] Preferably, the mass concentration of polyethylene glycol in the cellulose regeneration solution is 0.5-2%.

[0014] Preferably, the mass ratio of the zeolite to the carboxymethyl cellulose is 3-14:20.

[0015] Preferably, the dialysis is performed using ultrapure water or ethanol.

[0016] Another object of the embodiment of the present invention is to provide a zeolite-loaded regenerated cellulose aerogel hemostatic material, which is prepared by the above preparation method.

[0017] The present invention provides a method for preparing a zeolite-loaded regenerated cellulose aerogel hemostatic material. The method utilizes high-molecular-weight carboxymethyl cellulose to dissolve under alkaline conditions by disrupting hydrogen bonding interactions, then blends with zeolite to form a stable homogenate. In a coagulation bath system, hydrogen bonding restructures the fibers to form a three-dimensional network structure of a hydrogel. The cations in the zeolite are chelated by the carboxyl groups of the carboxymethyl groups, so that the zeolite is evenly loaded between the fibers. The zeolite-loaded regenerated cellulose aerogel material is then freeze-dried to form the zeolite-loaded regenerated cellulose aerogel material.

[0018] Compared with traditional zeolite-based hemostatic materials, the embodiments of the present invention form regenerated fiber aerogel materials through green solvents. Zeolite is stably combined in the three-dimensional structure of the aerogel and has excellent hemostatic properties. Its hemostatic mechanism includes: physical adsorption, when the aerogel acts on the bleeding site, it can quickly adsorb the liquid components in the blood and enrich blood cells and other components on the interface; interface stimulation, the electric potential carried by the zeolite can stimulate blood cells to trigger the coagulation cascade reaction; thermal stimulation: the three-dimensional structure of the aerogel can quickly disperse the heat generated by the zeolite to prevent the occurrence of thermal damage. At the same time, this heat can enhance platelet function. The hemostatic material has the advantages of fast hemostasis, mild exothermic reaction, and biosafety. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 This is a photo of the zeolite-loaded regenerated cellulose aerogel hemostatic material provided in Example 1 of the present invention;

[0020] Figure 2 SEM images of the zeolite-loaded regenerated cellulose aerogel hemostatic material provided in Example 1 of the present invention (left side is a cross-sectional scanning electron microscope image, right side is a surface scanning electron microscope image);

[0021] Figure 3 The in vitro coagulation performance evaluation results of the zeolite-loaded regenerated cellulose aerogel hemostatic materials prepared in Examples 1-3 of the present invention are shown (data bar graph on the left, actual display image on the right);

[0022] Figure 4 The blood compatibility test results of the zeolite-loaded regenerated cellulose aerogel hemostatic material prepared in Example 1 of the present invention (the inset is a real-life demonstration);

[0023] Figure 5 This is a graph showing the heat release evaluation during the in vivo hemostasis process of the zeolite-loaded regenerated cellulose aerogel hemostatic material prepared in Example 1 of the present invention;

[0024] Figure 6 These are the in vivo hemostatic performance evaluation results of the zeolite-loaded regenerated cellulose aerogel hemostatic material prepared in Example 1 of the present invention. DETAILED DESCRIPTION

[0025] In order to make the purpose, technical solutions and advantages of the present invention more clearly understood, the present invention will be further described in detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not intended to limit the present invention.

[0026] A method for preparing a zeolite-loaded regenerated cellulose aerogel hemostatic material comprises the following steps:

[0027] (1) Dissolving carboxymethylated cellulose and polyethylene glycol in an alkaline solution to obtain a cellulose regeneration solution, then adding zeolite and stirring evenly to obtain a white stable zeolite-regenerated cellulose slurry;

[0028] (2) placing the zeolite-regenerated cellulose slurry in an acid / alcohol coagulation bath of a certain concentration for fiber regeneration to obtain a zeolite-regenerated cellulose hydrogel, wherein the acid / alcohol coagulation bath is a mixture of an acid solution and an alcohol, wherein the acid solution is 10% by mass sulfuric acid and the alcohol is 50% by mass ethanol, and the mass ratio of the acid solution to the alcohol is 8:92;

[0029] (3) The zeolite-regenerated cellulose hydrogel was dialyzed to neutrality and the salt was removed, and then freeze-dried to obtain the zeolite-loaded regenerated cellulose aerogel hemostatic material, which was then frozen in liquid nitrogen to make its structure more compact.

[0030] Wherein, the degree of substitution of the carboxymethyl cellulose is 0.10-0.50, preferably 0.16; the mass concentration of the carboxymethyl cellulose is 3-10%, preferably 7%; the alkaline solution includes but is not limited to sodium hydroxide and potassium hydroxide, preferably sodium hydroxide;

[0031] The mass concentration of the polyethylene glycol is 0.5%-2%, preferably 1.5%;

[0032] The mass ratio of the zeolite to the carboxymethyl cellulose is 3-14:20, preferably 3:20;

[0033] The dialysis method is ultrapure water washing or ethanol washing, preferably ultrapure water washing.

[0034] The specific implementation of the present invention is described in detail below with reference to specific embodiments.

[0035] Among them, the zeolite used in the embodiments of the present invention was purchased from Taobao-Green New Source Environmental Protection (Zhengzhou, Henan) powdered molecular sieve (Ca-5A); carboxymethyl cellulose was purchased from Shandong Senxin Environmental Protection Technology Co., Ltd.; the average molecular weight of polyethylene glycol was 8000, purchased from Shanghai Titan Technology Co., Ltd.; PBS buffer solution has a pH of 7.4 and includes the following components: Na2HPO4 (anhydrous disodium hydrogen phosphate): Shanghai Titan Technology Co., Ltd., KH2PO4 (potassium dihydrogen phosphate): Shanghai Titan Technology Co., Ltd., KCl (potassium chloride): Shanghai McLean Biochemical Technology Co., Ltd., sodium chloride (NaCl): Sinopharm Chemical Reagent Co., Ltd.

[0036] Example 1: A method for preparing a zeolite-loaded regenerated cellulose aerogel hemostatic material, comprising the following steps:

[0037] (1) 3 g of carboxymethylated cellulose with a carboxymethyl substitution degree of 0.16 and 1.5 g of polyethylene glycol were dissolved in 95.5 g of 7% sodium hydroxide solution to obtain a 3% cellulose regeneration solution. 0.45 g of zeolite powder was then added and stirred to obtain a zeolite-regenerated cellulose slurry.

[0038] (2) transferring the zeolite-regenerated cellulose slurry into a mold and regenerating the fibers in an acid / alcohol coagulation bath to form a zeolite-regenerated cellulose hydrogel;

[0039] (3) The zeolite-regenerated cellulose hydrogel is washed with water until neutral, and freeze-dried at low temperature to form a zeolite-loaded regenerated cellulose aerogel hemostatic material (the ratio of zeolite to carboxymethyl cellulose is 3:20), such as Figure 1 shown.

[0040] The aerogel hemostatic material prepared in Example 1 was observed using a scanning electron microscope, and the structure was as follows: Figure 2 As shown, according to Figure 2 It can be seen that the interior of the aerogel hemostatic material is a porous three-dimensional network structure, and the zeolite is evenly distributed on the fiber surface.

[0041] Example 2: A method for preparing a zeolite-loaded regenerated cellulose aerogel hemostatic material, comprising the following steps:

[0042] (1) 3 g of carboxymethylated cellulose with a carboxymethyl substitution degree of 0.16 and 1.5 g of polyethylene glycol were dissolved in 95.5 g of 7% sodium hydroxide solution to obtain a 3% cellulose regeneration solution. 1 g of zeolite powder was then added and stirred to obtain a zeolite-regenerated cellulose slurry.

[0043] (2) transferring the zeolite-regenerated cellulose slurry into a mold and regenerating the fibers in an acid / alcohol coagulation bath to form a zeolite-regenerated cellulose hydrogel;

[0044] (3) The zeolite-regenerated cellulose hydrogel was washed with water until neutral and freeze-dried at low temperature to form a zeolite-loaded regenerated cellulose aerogel hemostatic material (the ratio of zeolite to carboxymethyl cellulose was 1:3).

[0045] Example 3: A method for preparing a zeolite-loaded regenerated cellulose aerogel hemostatic material, comprising the following steps:

[0046] (1) 3 g of carboxymethylated cellulose with a carboxymethyl substitution degree of 0.16 and 1.5 g of polyethylene glycol were dissolved in 95.5 g of 7% sodium hydroxide solution to obtain a 3% cellulose regeneration solution. 2 g of zeolite powder was then added and stirred to obtain a zeolite-regenerated cellulose slurry.

[0047] (2) transferring the zeolite-regenerated cellulose slurry into a mold and regenerating the fibers in an acid / alcohol coagulation bath to form a zeolite-regenerated cellulose hydrogel;

[0048] (3) The zeolite-regenerated cellulose hydrogel was washed with water until neutral and freeze-dried at low temperature to form a zeolite-loaded regenerated cellulose aerogel hemostatic material (the ratio of zeolite to carboxymethyl cellulose was 2:3).

[0049] Performance Analysis:

[0050] 1. The zeolite-loaded regenerated cellulose aerogel hemostatic material, medical cotton, and commercial Quikclot Combat Guaze prepared in Examples 1-3 were subjected to in vitro blood coagulation performance evaluation (BCI) comparison. The specific method is as follows:

[0051] Several milliliters of fresh rabbit blood were taken and anticoagulant was added (volume ratio of 9:1). A 0.2 M calcium chloride solution was prepared and allowed to stand. 100 mg of material was weighed and placed at the bottom of a centrifuge tube and preheated at 37°C for 5 minutes. Then 100 μL of anticoagulant blood was added, along with 20 μL of CaCl2 solution. The beaker was then placed at 37°C for 10 minutes. 25 mL of deionized water was then added, and the mixture was placed at 37°C for another 5 minutes. Finally, 200 μL of the supernatant was taken and the absorbance (Abs) value was measured using a visible light spectrometer (wavelength 540 nm). A blank control group and a positive control group (medical hemostatic cotton) were set up during the test. Each sample was tested 6 times and the average value was obtained.

[0052] The BCI calculation formula is: ;

[0053] Generally speaking, the smaller the BCI value, the better the hemostatic effect of the hemostatic material;

[0054] The result is as follows Figure 3 As shown, according to Figure 3 It can be seen that the zeolite-loaded regenerated cellulose aerogel hemostatic materials prepared in Examples 1-3 have excellent in vitro coagulation performance.

[0055] 2. The zeolite-loaded regenerated cellulose aerogel hemostatic material prepared in Example 1, medical absorbent cotton, and commercial Quikclot Combat Guaze were subjected to a blood compatibility test for comparison. The specific process is as follows:

[0056] Citrated whole blood was centrifuged at 2000 rpm for 10 minutes, and the erythrocyte sediment was washed three times with PBS. Then, 200 μL of pure erythrocytes were mixed with 9.8 mL of PBS to obtain a 2 v / v% erythrocyte suspension. 10 mg of zeolite-loaded regenerated cellulose aerogel hemostatic material, Quikclot, and cotton were placed in test tubes containing 1 mL of erythrocyte suspension and incubated at 37°C for 1 hour. At the same time, 50 μL of PBS and 2 wt% TritonX-100 were added to 1 mL of erythrocyte suspension as negative and positive controls, respectively. After that, all test tubes were further centrifuged at 2000 rpm for 10 minutes. Finally, the absorbance of the supernatant was measured at 540 nm, and six measurements were performed for each group.

[0057] The formula for calculating the hemolytic ratio (HR) is: HR (%) = [(OD s -OD p ) / (OD t -OD p )] * 100%;

[0058] OD s is the absorbance of the sample, OD t is the absorbance of 2 wt% TritonX-100, OD p is the absorbance of PBS;

[0059] The result is as follows Figure 4 As shown, according to Figure 4 It can be seen that the zeolite-loaded regenerated cellulose aerogel hemostatic material prepared in Example 1 does not produce hemolysis at a concentration of 10 mg / ml.

[0060] 3. The heat release evaluation during in vivo hemostasis of the zeolite-loaded regenerated cellulose aerogel hemostatic material prepared in Example 1 and the commercial Quiclot combat gauze was compared. The specific steps are as follows:

[0061] At room temperature, the heat release capacity of different materials during the hemostasis process in mice was measured using an infrared thermal imager. IR imaging was performed during the hemostasis process in a mouse liver injury model, and the highest heat release temperature was recorded until the temperature almost stopped changing.

[0062] The result is as follows Figure 5 As shown, according to Figure 5 It can be seen that the zeolite-loaded regenerated cellulose aerogel hemostatic material prepared in Example 1 reduces the risk of thermal injury.

[0063] 4. The in vivo hemostatic performance of the zeolite-loaded regenerated cellulose aerogel hemostatic material, medical absorbent cotton, and commercial Quiclot combat gauze prepared in Example 1 was evaluated and compared. The specific steps are as follows:

[0064] A rat femoral artery rupture bleeding model was used. Sprague-Dawley rats were anesthetized and fixed on their backs with their abdomens facing upwards. The hair at the base of the hind legs was shaved and the base of the hind legs was disinfected with 75% medical alcohol. The femoral artery was then located and an incision was made at the epidermis of the femoral artery at the base of the thigh. The femoral artery was cut with surgical scissors and allowed to bleed freely for 5 seconds. The floating blood on the wound was then gently wiped away. A hemostatic material was pressed on the bleeding wound and a timer was started. The hemostatic material was gently removed every 30 seconds to observe whether bleeding continued. If bleeding continued, the pressure was continued until bleeding stopped. After hemostasis was complete, each material was weighed and the blood loss was calculated. The experiment was repeated 6 times for each sample.

[0065] The result is as follows Figure 6 As shown, according to Figure 6 It can be seen that the zeolite-loaded regenerated cellulose aerogel hemostatic material prepared in Example 1 has excellent in vivo hemostatic performance.

[0066] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A method for preparing a zeolite-loaded regenerated cellulose aerogel hemostatic material, characterized in that: The following steps are involved: Dissolve carboxymethylated cellulose and polyethylene glycol in an alkaline solution to obtain a cellulose regeneration solution, then add zeolite and stir evenly to obtain a white stable zeolite-regenerated cellulose slurry; Placing the zeolite-cellulose regenerated cellulose homogenate in an acid / alcohol coagulation bath for fiber regeneration to obtain a zeolite-regenerated cellulose hydrogel, wherein the acid / alcohol coagulation bath is a mixture of an acid solution and an alcohol, wherein the acid solution is 10% by mass sulfuric acid and the alcohol is 50% by mass ethanol, and the mass ratio of the acid solution to the alcohol is 8:92; The zeolite-regenerated cellulose hydrogel is dialyzed to neutrality and salt is removed, and then freeze-dried to obtain a zeolite-loaded regenerated cellulose aerogel hemostatic material; The carboxymethyl degree of substitution of the carboxymethyl cellulose is 0.16; The alkaline solution is sodium hydroxide or potassium hydroxide; The mass concentration of carboxymethyl cellulose in the cellulose regeneration solution is 3-10%; The mass concentration of polyethylene glycol in the cellulose regeneration solution is 0.5-2%; The mass ratio of the zeolite to the carboxymethyl cellulose is 3-14:

20.

2. The method for preparing the zeolite-loaded regenerated cellulose aerogel hemostatic material according to claim 1, characterized in that: The dialysis is performed by washing with ultrapure water or ethanol.

3. A zeolite-loaded regenerated cellulose aerogel hemostatic material, characterized in that: The invention discloses a method for preparing the present invention according to claim 1 or 2.

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