Absorbable hemostatic bone wax and preparation method thereof

Abstract

The present invention provides an absorbable hemostatic bone wax and a preparation method thereof, belonging to the technical field of hemostatic materials. The wax comprises poloxamer and a random copolymer of ethylene oxide and propylene oxide, with a mass ratio of 3-7:3-7. The absorbable hemostatic bone wax is a waxy solid material with good plasticity and adhesion. It is used to control bleeding from bone wounds and is completely degraded and absorbed within one month after use, avoiding the risk of bone nonunion and infection caused by foreign body space occupation.

Description

Technical Field

[0001] The invention relates to the technical field of hemostatic materials, and in particular to absorbable hemostatic bone wax and a preparation method thereof. Background Art

[0002] Bone wax is commonly used clinically to stop bleeding from cancellous bone wounds. Bone wax is a mixture of sterilized beeswax and petroleum jelly. It is white or light yellow in color and has excellent softening properties. It becomes malleable after being rubbed by hand. Bone wax works by physically blocking bleeding from bone capillaries. However, issues with bone wax include: It is difficult to degrade and absorb in the human body after use, remaining as a foreign body for a long time, hindering bone healing and increasing the risk of postoperative infection.

[0003] Ideal orthopedic hemostatic materials should have the following characteristics: ① They can be degraded and absorbed in the human body without hindering bone healing; ② They have good biocompatibility, no toxic side effects, and little tissue reaction; ③ They have good adhesion and can firmly adhere to the bone wound surface to quickly and effectively stop bleeding; ④ They have good plasticity and can be used for bone wounds of various shapes. Summary of the Invention

[0004] The purpose of the present invention is to propose an absorbable hemostatic bone wax and a preparation method thereof. The wax is a waxy solid material with good plasticity and adhesion. It is used to control bleeding from bone wounds. After use, it can be completely degraded and absorbed within one month, avoiding the risk of bone non-union and infection caused by foreign body occupation.

[0005] The technical solution of the present invention is achieved as follows:

[0006] The invention provides an absorbable hemostatic bone wax, which comprises poloxamer and an ethylene oxide / propylene oxide random copolymer, wherein the mass ratio of the two copolymers is 3-7:3-7.

[0007] As a further improvement of the present invention, the poloxamer comprises poloxamer 188 and poloxamer 407 in a mass ratio of 5-10:3-5.

[0008] As a further improvement of the present invention, the average molecular weight of the poloxamer 188 is 7680-9510, and the ethylene oxide content is 79.9%-83.7%; the average molecular weight of the poloxamer 407 is 9840-14600, and the ethylene oxide content is 71.5%-74.9%; and the average molecular weight of the ethylene oxide propylene oxide random copolymer is 12000, and the ethylene oxide content is 70-80%.

[0009] As a further improvement of the present invention, it also includes a sodium alginate-soy protein hydrolyzed porous hydrogel, the mass of which is 10-20wt% of the poloxamer.

[0010] As a further improvement of the present invention, the preparation method of the sodium alginate-soy protein hydrolyzed porous hydrogel is as follows:

[0011] S1. Dissolve soy protein hydrolyzate in water to obtain a protein solution;

[0012] S2. dissolving sodium alginate in water to obtain a sodium alginate solution;

[0013] S3. The protein solution obtained in step S1 and the sodium alginate solution obtained in step S2 are evenly mixed, a metal salt solution is added, solidified at room temperature, freeze-dried, and ground to obtain a sodium alginate-soy protein hydrolyzed porous hydrogel.

[0014] The present invention further protects a method for preparing the absorbable hemostatic bone wax, comprising the following steps:

[0015] (1) Poloxamer and ethylene oxide propylene oxide random copolymer are heated in a double-layer glass reactor and completely dissolved;

[0016] (2) Keep the temperature constant and stir to make it disperse evenly;

[0017] (3) Keep the temperature constant, turn off the stirring, and let it stand until most of the bubbles are eliminated;

[0018] (4) Keep the temperature constant and vacuum to remove bubbles;

[0019] (5) Pour the mixed liquid into a preheated mold;

[0020] (6) The mold is quickly transferred to a freeze dryer for freeze molding, packaging, and sterilization to obtain absorbable hemostatic bone wax.

[0021] As a further improvement of the present invention, the heating temperature in step (1) is 75-85° C. and the heating time is 1-2 h; the stirring speed in step (2) is 500-700 r / min and the stirring time is 20-30 min.

[0022] As a further improvement of the present invention, the standing time in step (3) is 100-140 min; the vacuum degree in step (4) is drawn to -0.05 to -0.1 MPa, and the time for removing bubbles is 40-60 min.

[0023] As a further improvement of the present invention, the freezing temperature in step (5) is -40 to -50°C and the freezing time is 20-40 minutes.

[0024] The present invention further protects a method for preparing the absorbable hemostatic bone wax, comprising the following steps:

[0025] (1) Poloxamer and ethylene oxide propylene oxide random copolymer were heated in a double-layer glass reactor at 75-85°C for 1-2 hours to completely dissolve;

[0026] (2) Keeping the temperature constant, add sodium alginate-soy protein hydrolyzed porous hydrogel and stir at 500-700 rpm for 20-30 seconds to disperse it evenly;

[0027] (3) Keep the temperature constant, turn off the stirring, and let it stand for 100-140 minutes to eliminate most of the bubbles;

[0028] (4) Keeping the temperature constant, reduce the vacuum degree to -0.05 to -0.1 MPa and remove bubbles for 40-60 minutes;

[0029] (5) Pour the mixed liquid into a preheated mold;

[0030] (6) The mold is quickly transferred to a freeze dryer at -40 to -50°C and frozen for 20-40 minutes to form, package, and sterilize to obtain absorbable hemostatic bone wax.

[0031] The present invention has the following beneficial effects: the absorbable hemostatic bone wax of the present invention is a waxy solid material with good plasticity and adhesion, and is used to control bleeding from bone wounds. After use, it can be completely degraded and absorbed within one month, avoiding the risk of bone non-union and infection caused by foreign body occupation. DETAILED DESCRIPTION

[0032] The following is a clear and complete description of the technical solutions in the embodiments of the present invention. Obviously, the embodiments described are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making any creative efforts are within the scope of protection of the present invention.

[0033] Preparation Example 1 Preparation of Sodium Alginate-Soybean Hydrolyzed Protein Porous Hydrogel

[0034] Here’s how:

[0035] S1. The soy protein hydrolyzate was dissolved in water to obtain a protein solution having a concentration of 3wt%;

[0036] S2. Dissolve sodium alginate in water to obtain a 5 wt% sodium alginate solution;

[0037] S3. The protein solution obtained in step S1 and the sodium alginate solution obtained in step S2 were evenly mixed, 2 wt% calcium chloride solution was added, solidified at room temperature for 30 min, freeze-dried, and ground to obtain a sodium alginate-soy protein hydrolyzed porous hydrogel.

[0038] Preparation Example 2 Preparation of Sodium Alginate-Soybean Hydrolyzed Protein Porous Hydrogel

[0039] Here’s how:

[0040] S1. The soy protein hydrolyzate was dissolved in water to obtain a protein solution having a concentration of 5wt%;

[0041] S2. Dissolving sodium alginate in water to obtain a 7 wt % sodium alginate solution;

[0042] S3. The protein solution obtained in step S1 and the sodium alginate solution obtained in step S2 were evenly mixed, 3 wt% calcium chloride solution was added, solidified at room temperature for 30 min, freeze-dried, and ground to obtain a sodium alginate-soy protein hydrolyzed porous hydrogel.

[0043] Preparation Example 3 Preparation of Sodium Alginate-Soybean Hydrolyzed Protein Porous Hydrogel

[0044] Here’s how:

[0045] S1. The soy protein hydrolyzate was dissolved in water to obtain a protein solution having a concentration of 4wt%;

[0046] S2. Dissolving sodium alginate in water to obtain a 6 wt% sodium alginate solution;

[0047] S3. The protein solution obtained in step S1 and the sodium alginate solution obtained in step S2 were evenly mixed, 2.5 wt% calcium chloride solution was added, solidified at room temperature for 30 min, freeze-dried, and ground to obtain a sodium alginate-soy protein hydrolyzed porous hydrogel.

[0048] Example 1

[0049] This embodiment provides an absorbable hemostatic bone wax comprising a poloxamer and an ethylene oxide-propylene oxide random copolymer in a mass ratio of 3:3. The poloxamer comprises poloxamer 188 and poloxamer 407 in a mass ratio of 5:3. Poloxamer 188 has an average molecular weight of 7680 and an ethylene oxide content of 79.9%; poloxamer 407 has an average molecular weight of 9840 and an ethylene oxide content of 71.5%; and the ethylene oxide-propylene oxide random copolymer has an average molecular weight of 12000 and an ethylene oxide content of 70%.

[0050] The preparation method comprises the following steps:

[0051] (1) Poloxamer and ethylene oxide propylene oxide random copolymer were placed in a double-layer glass reactor, heated to 75°C, and stirred to dissolve for 1 hour;

[0052] (2) Keep the temperature constant and stir at 500 r / min for 20 min;

[0053] (3) Keep the temperature constant, turn off the stirring, and let it stand for 100 minutes to eliminate most of the bubbles;

[0054] (4) Maintaining the temperature constant, evacuate to -0.05 MPa and remove bubbles for 40 minutes;

[0055] (5) Pour the mixed liquid into a preheated mold;

[0056] (6) The mold was quickly transferred to a freeze dryer, frozen at -40°C for 20 minutes, packaged, and sterilized to obtain absorbable hemostatic bone wax.

[0057] Example 2

[0058] This embodiment provides an absorbable hemostatic bone wax comprising a poloxamer and an ethylene oxide-propylene oxide random copolymer in a mass ratio of 3:7. The poloxamer comprises poloxamer 188 and poloxamer 407 in a mass ratio of 10:5. Poloxamer 188 has an average molecular weight of 9510 and an ethylene oxide content of 83.7%. Poloxamer 407 has an average molecular weight of 14600 and an ethylene oxide content of 74.9%. The ethylene oxide-propylene oxide random copolymer has an average molecular weight of 12000 and an ethylene oxide content of 80%.

[0059] The preparation method comprises the following steps:

[0060] (1) Poloxamer and ethylene oxide propylene oxide random copolymer were heated to 85°C in a double-layer glass reactor and stirred for 2 h to dissolve;

[0061] (2) Keep the temperature constant and stir at 500-700 rpm for 30 min;

[0062] (3) Keep the temperature constant, turn off the stirring, and let it stand for 140 minutes to eliminate most of the bubbles;

[0063] (4) Maintaining the temperature constant, evacuate to -0.1 MPa and remove bubbles for 60 minutes;

[0064] (5) Pour the mixed liquid into a preheated mold;

[0065] (6) The mold was quickly transferred to a freeze dryer, frozen at -50°C for 40 minutes, packaged, and sterilized to obtain absorbable hemostatic bone wax.

[0066] Example 3

[0067] This embodiment provides an absorbable hemostatic bone wax comprising a poloxamer and an ethylene oxide-propylene oxide random copolymer in a mass ratio of 7:3. The poloxamer comprises poloxamer 188 and poloxamer 407 in a mass ratio of 7:4. Poloxamer 188 has an average molecular weight of 8210 and an ethylene oxide content of 81%; poloxamer 407 has an average molecular weight of 12600 and an ethylene oxide content of 72.5%; and the ethylene oxide-propylene oxide random copolymer has an average molecular weight of 12000 and an ethylene oxide content of 75%.

[0068] The preparation method comprises the following steps:

[0069] (1) Poloxamer and ethylene oxide propylene oxide random copolymer were heated to 80°C in a double-layer glass reactor and stirred for 1.5 hours to dissolve;

[0070] (2) Keep the temperature constant and stir at 600 r / min for 25 min;

[0071] (3) Keep the temperature constant, turn off the stirring, and let it stand for 120 minutes to eliminate most of the bubbles;

[0072] (4) Keep the temperature constant, evacuate to -0.1 MPa, and remove bubbles for 50 minutes;

[0073] (5) Pour the mixed liquid into a preheated mold;

[0074] (6) The mold was quickly transferred to a freeze dryer, frozen at -45°C for 30 minutes, packaged, and sterilized to obtain absorbable hemostatic bone wax.

[0075] Example 4

[0076] Compared with Example 3, the difference is that the sodium alginate-soy protein hydrolyzed porous hydrogel prepared in Preparation Example 1 is also included, and the mass of the sodium alginate-soy protein hydrolyzed porous hydrogel is 10 wt % of the poloxamer.

[0077] (1) Poloxamer and ethylene oxide propylene oxide random copolymer were heated to 80°C in a double-layer glass reactor and stirred for 1.5 hours to dissolve;

[0078] (2) Keeping the temperature constant, add sodium alginate-soy protein hydrolyzed porous hydrogel and stir at 600 rpm for 25 min;

[0079] (3) Keep the temperature constant, turn off the stirring, and let it stand for 120 minutes to eliminate most of the bubbles;

[0080] (4) Keep the temperature constant, evacuate to -0.1 MPa, and remove bubbles for 50 minutes;

[0081] (5) Pour the mixed liquid into a preheated mold;

[0082] (6) The mold was quickly transferred to a freeze dryer, frozen at -45°C for 30 minutes, packaged, and sterilized to obtain absorbable hemostatic bone wax.

[0083] Example 5

[0084] Compared with Example 4, the difference is that the mass of the sodium alginate-soy protein hydrolyzed porous hydrogel is 20 wt % of the poloxamer.

[0085] Example 6

[0086] Compared with Example 4, the difference is that the mass of the sodium alginate-soy protein hydrolyzed porous hydrogel is 15 wt % of the poloxamer.

[0087] Test Example 1

[0088] The absorbable hemostatic bone wax prepared in Examples 1-6 of the present invention was subjected to performance testing.

[0089] 1. Degradation performance

[0090] Muscle Implantation Test: After 14 days of implantation, the sample completely degraded with no tissue reaction. The sample was implanted into rabbit muscle tissue to evaluate its potential for irritation and toxicity. This test was conducted in accordance with the national standard GB / T 16886.6-2015, "Biological Evaluation of Medical Devices - Part 6: Post-implantation Local Reactions Test."

[0091] The test samples and control samples were implanted into the muscle tissue on both sides of the back of rabbits, respectively. The animals were killed 2 days, 1 week, and 2 weeks after implantation. The implantation sites were observed with the naked eye, and sufficient unaffected muscle tissue including the implanted samples and the surrounding area was excised for HE staining. The inflammatory reaction around the samples, the cyst cavity, and the cyst wall formation were observed and evaluated under a light microscope.

[0092] Under the conditions of this study, visual inspection revealed no abnormalities in the tissue structure of the test sample's absorbable hemostatic bone wax, nor in the muscle tissue of the control sample's implant site. Histopathological examination revealed no irritation in either the test or control sample at two days, one week, or two weeks. Fourteen days after implantation, no sample residue was observed in either the test or control sample, both visually and microscopically.

[0093] 2. Cytotoxicity

[0094] Samples were extracted at a ratio of 0.2 g / mL in serum-containing culture medium at (37 ± 1)°C for (24 ± 2) hours. The extract was used as the test fluid and the test was performed according to the method specified in GB / T 16886.5-2017, "Biological Evaluation of Medical Devices - Part 5: In Vitro Cytotoxicity Tests." Cytotoxicity results were expressed as the relative cell proliferation rate (RGR). A higher RGR indicates lower cytotoxicity and better product safety.

[0095] Results: The relative growth rates (RGR) of the absorbable hemostatic bone waxes prepared in Examples 1-6 were all above 75%, and the cytotoxicity was no greater than level one. The results are shown in Table 1 for details.

[0096] Table 1

[0097] Group Relative proliferation rate (RGR) Example 1 97％ Example 2 95％ Example 3 98％ Example 4 101％ Example 5 97％ Example 6 93％

[0098] 3. Adhesion

[0099] Weigh 1.0g of sample, knead thoroughly until there are no particles, roll it into a ball and place it between two adhesion test plates. Place two 1mm diameter steel wires between the test plates and press the sample into a round cake with a thickness of 1mm. Leave it at room temperature for 30 minutes. Fix one test plate to the test stand and hang a 500g weight from the other test plate. The weight should remain there for 30 minutes without falling off.

[0100] Results: The absorbable hemostatic bone waxes prepared in Examples 1-6 did not fall off within 30 minutes, and the absorbable hemostatic bone waxes prepared in Examples 4-6 did not fall off within 90 minutes.

[0101] 4. Plasticity

[0102] Plasticity refers to a material's ability to deform plastically before breaking under stress. Referring to the determination method for dental zinc phosphate cement in ISO 1566-1978 Dental zinc phosphate cements, the plasticity of absorbable hemostatic bone wax is defined as the diameter of the circle formed by rolling a 2.5g test sample into a ball and pressing a 1kg weight onto the ball through a plastic plate. The diameter of the circle represents the plasticity of the sample; the larger the diameter, the better the plasticity. Three samples of the same formula were tested, and the vertical and horizontal diameters of each circle were measured, and the average value was taken.

[0103] Results: The plasticity of the absorbable hemostatic bone wax prepared in Examples 1-6 was 21 mm, which was greater than the plasticity of the non-absorbable bone wax of 18 mm.

[0104] 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, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. An absorbable hemostatic bone wax, characterized in that: The invention comprises poloxamer and ethylene oxide propylene oxide random copolymer, the mass ratio of the two being 3-7:3-7, and also comprises sodium alginate-soy protein hydrolyzed porous hydrogel, the mass of which is 10-20wt% of the poloxamer. The preparation method of the sodium alginate-soy protein hydrolyzed porous hydrogel is as follows: S1. Dissolve soy protein hydrolyzate in water to obtain a protein solution; S2. dissolving sodium alginate in water to obtain a sodium alginate solution; S3. The protein solution obtained in step S1 and the sodium alginate solution obtained in step S2 are evenly mixed, a metal salt solution is added, solidified at room temperature, freeze-dried, and ground to obtain a sodium alginate-soy protein hydrolyzed porous hydrogel.

2. The absorbable hemostatic bone wax according to claim 1, characterized in that: The poloxamer includes poloxamer 188 and poloxamer 407, with a mass ratio of 5-10:3-5.

3. The absorbable hemostatic bone wax according to claim 1, characterized in that: The average molecular weight of the poloxamer 188 is 7680-9510, and the ethylene oxide content is 79.9%-83.7%; the average molecular weight of the poloxamer 407 is 9840-14600, and the ethylene oxide content is 71.5%-74.9%; the average molecular weight of the ethylene oxide propylene oxide random copolymer is 12000, and the ethylene oxide content is 70-80%.

4. A method for preparing the absorbable hemostatic bone wax according to any one of claims 1 to 3, characterized in that: The following steps are involved: (1) Poloxamer and ethylene oxide propylene oxide random copolymer are heated in a double-layer glass reactor at 75-85°C for 1-2 hours to completely dissolve; (2) Keeping the temperature constant, add sodium alginate-soy protein hydrolyzed porous hydrogel and stir at 500-700 r / min for 20-30 min to make it evenly dispersed; (3) Keep the temperature constant, turn off the stirring, and let it stand for 100-140 minutes to eliminate most of the bubbles; (4) Keep the temperature constant, reduce the vacuum to -0.05 to -0.1 MPa, and remove bubbles for 40-60 minutes; (5) Pour the mixed liquid into the preheated mold; (6) The mold is quickly transferred to a freeze dryer at -40 to -50°C and frozen for 20-40 minutes to form, package, and sterilize to obtain absorbable hemostatic bone wax.