Tamponade biological material for hemostasis and preparation method therefor

Abstract

A tamponade biological material for hemostasis and a preparation method therefor. The tamponade biological material is obtained by pretreating, crosslinking, and post-treating a small intestinal submucosal material. The tamponade biological material has good water resistance, a low swelling rate, and an appropriate degradation speed; it is degradable and not prone to adhesion and curling, thereby avoiding wound traction and reducing patient pain, so it is suitable for hemostasis in various situations, and has good market application prospects.

Description

A filling biomaterial for hemostasis and a preparation method thereof TECHNICAL FIELD

[0001] The present application belongs to the field of biomaterials, and particularly relates to a filling biomaterial for hemostasis and a preparation method thereof. BACKGROUND

[0002] Traditional clinical hemostasis methods include suturing, compression and the like, but when facing surgeries such as deep interstitial spaces and large wounds, the traditional hemostasis methods are difficult to be effective, in addition, the traditional hemostasis methods also cannot solve the problem of secondary bleeding caused by postoperative traction and the like, therefore, hemostasis materials emerge as the times require. Among them, the hemostasis effect of pure collagen protein biomaterials is very obvious. Collagen protein can make platelets aggregate, thereby stimulating platelets to release coagulation factors, thereby achieving hemostasis effect. However, such materials have poor water resistance, are easy to swell when encountering water, and have too fast degradation speed. Therefore, it is necessary to develop a new hemostasis material. TECHNICAL SOLUTION

[0003] The present application provides a filling biomaterial for hemostasis in the first aspect, which is obtained by pretreating, cross-linking and post-treating the small intestinal submucosa.

[0004] The present application also provides a preparation method of the filling biomaterial for hemostasis, comprising the following steps:

[0005] (1) Pretreatment: the small intestinal tissue of a pig is mechanically stripped to obtain small intestinal submucosa; then the pretreated protein material is obtained by defatting and deantigenization;

[0006] (2) Cross-linking: the protein material obtained in step (1) is cross-linked and modified in a cross-linking treatment solution to obtain a modified protein material; wherein the cross-linking treatment solution is composed of a catechol solution, a potassium persulfate solution, a sodium periodate solution and a chitosan quaternary ammonium salt;

[0007] (3) Post-treatment: the modified protein material obtained in step (2) is added into an alkali solution to adjust the pH value to alkaline, and the reaction is carried out for 24-36 hours; then the pH value is adjusted to acid or neutral, and the solid membrane layer is filtered, washed with water and freeze-dried to obtain the filling biomaterial.

[0008] Preferably, the defatting in step (1) is specifically as follows: the small intestinal submucosa, isopropyl alcohol and anhydrous ethanol are mixed according to a ratio of 1g:5-10mL:0.1-1mL, then extracted and filtered, the solid is taken out and washed, and the operation is completed.

[0009] Preferably, the deantigenization in step (1) is specifically as follows: the defatted small intestinal submucosa is extracted and filtered in a 3-4wt% sodium chloride alkaline aqueous solution, the solid is taken out and washed, and the operation is completed.

[0010] In step (1), when the defatted small intestinal submucosa is extracted in the 3-4wt% sodium chloride alkaline aqueous solution, the ratio of the defatted small intestinal submucosa to the 3-4wt% sodium chloride alkaline aqueous solution is (0.001-1g):(1-100mL).

[0011] Preferably, the volume ratio of the catechol solution, the potassium persulfate solution and the sodium periodate solution in step (2) is 1-2:0.1-0.5:0.1-0.2.

[0012] Preferably, the concentration of the chitosan quaternary ammonium salt in the crosslinking treatment solution is 5-10wt%.

[0013] Preferably, the concentration of the catechol solution is 4-8mg / mL; the concentration of the potassium persulfate solution is 10-15mg / mL; and the concentration of the sodium periodate solution is 0.1-1mg / mL.

[0014] Preferably, the crosslinking modification time in step (2) is 1h-1.5h.

[0015] Preferably, after step (3), the modified protein material obtained in step (2) is added into an alkaline solution to adjust the pH value to 8.2-9.5, and the reaction is carried out for 24-36h; then the pH value is adjusted to 6-7.5, and the filtration is carried out; the solid membrane layer is washed with water and then freeze-dried to obtain the tampon biomaterial. Advantages

[0016] The tampon biomaterial has good water resistance, low swelling rate and appropriate degradation speed, is degradable, not easy to adhere and not easy to curl, and can reduce the pain of patients, so it is suitable for hemostasis in various situations and has good market application prospect. Embodiments of the present application

[0017] The present application will be further described below in conjunction with specific examples. It should be understood that these examples are only used to illustrate the present application and not used to limit the scope of the present application. In addition, it should be understood that those skilled in the art can make various modifications or changes to the present application after reading the content taught by the present application, and these equivalent forms also fall within the scope defined by the appended claims of the present application.

[0018] The substances used in the examples are commercially available. In the present application, the chitosan quaternary ammonium salt is purchased from Kanglang Biology, model KL10818.

[0019] Example 1

[0020] (1) Pretreatment: the small intestinal tissue of a pig is mechanically stripped to obtain a small intestinal submucosa; then the defatted and antigen-removed protein material is obtained after pretreatment;

[0021] Defatting: Small intestinal submucosa, isopropanol, anhydrous ethanol are mixed according to the ratio of 1g:6mL:0.7mL, and after 10h of extraction, filtration is performed, and the solid is washed with deionized water for 20-40min.

[0022] Deantigen: After defatting, the solid is added with 3wt% sodium chloride-containing sodium hydroxide solution at a ratio of 0.02:20mL, and after 10h of extraction, filtration is performed, and the solid is washed with deionized water for 20-40min.

[0023] (2) Crosslinking: The protein material obtained in step (1) is subjected to crosslinking modification in a crosslinking treatment solution for 1h to obtain a modified protein material; wherein the crosslinking treatment solution is composed of catechol solution, potassium persulfate solution, sodium periodate solution and chitosan quaternary ammonium salt; in the crosslinking treatment solution, the concentration of chitosan quaternary ammonium salt is 5wt%; the volume ratio of catechol solution, potassium persulfate solution and sodium periodate solution is 1:0.5:0.2. The crosslinking treatment solution is obtained by mixing catechol solution, potassium persulfate solution and sodium periodate solution, and then adding chitosan quaternary ammonium salt.

[0024] (3) Post-treatment: The modified protein material obtained in step (2) is added with sodium hydroxide aqueous solution to adjust the pH value to 9, so that the modified protein material is immersed in the sodium hydroxide aqueous solution, and the reaction is carried out for 24h; then acetic acid aqueous solution is added to adjust the pH value to 7.4, and filtration is performed, and after washing with water, the solid membrane layer is freeze-dried to obtain a filling biological material.

[0025] Example 2

[0026] The difference from Example 1 is that in step (2), during crosslinking, the volume ratio of catechol solution, potassium persulfate solution and sodium periodate solution is 1:0.4:0.1.

[0027] Comparative Example 1

[0028] The difference from Example 1 is that in step (2), during crosslinking, the crosslinking treatment solution is only composed of catechol solution, potassium persulfate solution and sodium periodate solution. Chitosan quaternary ammonium salt is not added.

[0029] Comparative Example 2

[0030] The difference from Example 1 is that in step (2), during crosslinking, the crosslinking treatment solution is only composed of potassium persulfate solution, sodium periodate solution and chitosan quaternary ammonium salt. Catechol solution is not added.

[0031] Comparative Example 3

[0032] The difference from Example 1 is that in step (2), during crosslinking, the volume ratio of catechol solution, potassium persulfate solution and sodium periodate solution is 1:0.9:0.6.

[0033] Comparative Example 4

[0034] The difference from Example 1 is that after step (3), the modified protein material obtained in step (2) is added to PBS buffer to adjust the pH value to 7.4, the modified protein material is immersed in the PBS buffer, and the reaction is carried out for 24 h, then filtered, washed with water, and freeze-dried to obtain the filling biological material.

[0035] Performance test

[0036] ①Porosity: The materials obtained in Examples 1-2 and Comparative Examples 1-4 were characterized by scanning electron microscopy to observe the cross-sectional morphology, and the porosity was calculated therefrom, and the results are shown in Table 1:

[0037] Table 1

[0038]

[0039] The greater the porosity, the better the adsorption performance of the material after implantation, and the more conducive to wound hemostasis and wound recovery.

[0040] ②Swelling rate: The materials obtained in Examples 1-2 and Comparative Examples 1-4 were cut into 1x1 cm pieces, and the mass at this time (m) was measured. The pieces were immersed in PBS buffer, and after 2 h, they were taken out, the surface liquid was absorbed with filter paper, and then weighed again, recorded as m1, and the swelling rate was (m1-m) / m x 100%, and the results are shown in Table 2:

[0041] Table 2

[0042]

[0043] The lower the swelling rate, the smaller the pressure on the tissue, and the more conducive to wound recovery and less likely to cause rebleeding.

[0044] ③Water resistance: Before and after the above swelling test, the compression stress was measured using an Elf3330 dynamic / static multifunctional biomechanical testing machine at room temperature, with a compression speed of 10 mm / min, and the results are shown in Table 3:

[0045] Table 3

[0046]

[0047] ④Thermal test: Healthy adult rabbits of either sex were taken, and the filling biological materials obtained in Example 2 and Comparative Example 3 were respectively extracted with physiological saline, and injected into the marginal ear vein of the rabbit, the anal temperature of the rabbit was detected, and the temperature difference before and 2 hours after injection was compared. The temperature difference of Example 2 was 0.16℃, and the temperature difference of Comparative Example 3 was 1.05℃, both of which were below 1.4℃.

Claims

1. A packing biomaterial for hemostasis, characterized by: The small intestinal submucosa is obtained by pretreatment, cross-linking and post-treatment.

2. A method for preparing a tampon biological material for hemostasis according to claim 1, comprising the following steps: (1) Pretreatment: the small intestinal tissue of a pig is mechanically stripped to obtain small intestinal submucosa; then the pretreated protein material is obtained by degreasing and deantigenization; (2) Crosslinking: the protein material obtained in step (1) is subjected to crosslinking modification in a crosslinking treatment solution to obtain a modified protein material; wherein, The cross-linking treatment solution is composed of a catechol solution, a potassium persulfate solution, a sodium periodate solution and a chitosan quaternary ammonium salt. (3) Post-treatment: the modified protein material obtained in step (2) is added into an alkaline solution to adjust the pH value to alkaline, and reacted for 24-36 h; then the pH value is adjusted to weakly acidic or neutral, and the solid membrane layer is filtered, washed with water and freeze-dried to obtain the tampon biological material.

3. The method of claim 2, wherein: The degreasing in step (1) is specifically as follows: the small intestinal submucosa, isopropyl alcohol and anhydrous ethanol are mixed at a ratio of 1 g:5-10 mL:0.1-1 mL, then extracted, filtered, the solid is taken out and washed, and the degreasing is completed.

4. The method of claim 2, wherein: The deantigenization in step (1) is specifically as follows: the degreased small intestinal submucosa is extracted in a 3-4 wt% sodium chloride alkaline aqueous solution, filtered, the solid is taken out and washed, and the deantigenization is completed.

5. The method of claim 2, wherein: In the cross-linking treatment solution in step (2), the volume ratio of the catechol solution, the potassium persulfate solution and the sodium periodate solution is 1-2:0.1-0.5:0.1-0.

2.

6. The production method according to claim 2 or 5, characterized by: In the cross-linking treatment solution, the concentration of the chitosan quaternary ammonium salt is 5-10 wt%.

7. The production method according to claim 2 or 5, characterized by: The concentration of the catechol solution is 4-8 mg / mL; the concentration of the potassium persulfate solution is 10-15 mg / mL; and the concentration of the sodium periodate solution is 0.1-1 mg / mL.

8. The method of claim 2, wherein: The cross-linking modification time in step (2) is 1 h-1.5 h.

9. The method of claim 2, wherein: In the post-treatment in step (3), the modified protein material obtained in step (2) is added into an alkaline solution to adjust the pH value to 8.2-9.5, and reacted for 24-36 h; then the pH value is adjusted to 6-7.5, and the solid membrane layer is filtered, washed with water and freeze-dried to obtain the tampon biological material.

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