CN105536029A - Preparation method for chitosan porous hemostatic sponge - Google Patents

Abstract

The invention discloses a preparation method of a chitosan porous hemostatic sponge, which comprises the following steps: step 1, preparation of a solution; step 2, preparation of a chitosan porous hemostatic sponge, taking chitosan solution, gelatin solution and glycerin solution According to the volume ratio of 3:3:1, fully stir and mix in the 25°C water bath condition to obtain a mixed milky white gel solution, then add calcium chloride solution, fully stir in the 25°C water bath condition for 30min, and then take the tannic acid solution Add, and continue to stir for 20min in a water bath at 25°C to obtain a mixed red gel solution, in which calcium chloride solution and tannic acid solution account for 0.5% and 0.01% respectively; The red gel liquid is poured into a mold, pre-frozen for more than 12 hours to form an ice cube, and then vacuum freeze-dried for 48 hours to obtain a chitosan porous hemostatic sponge. The beneficial effects are: simple preparation, high content of chitosan in the finished product, natural degradation, fast hemostasis speed, good antibacterial effect and the like.

Description

A kind of preparation method of chitosan porous hemostatic sponge

technical field

The invention belongs to the field of preparation of medical hemostatic materials, in particular to a preparation method of chitosan porous hemostatic sponge.

Background technique

Over the years, there have been more and more studies on hemostatic materials, most of which are natural products that can be degraded in vivo, such as fibrin glue, powdered collagen, gelatin sponge, but these materials have certain shortcomings, the source of fibrin glue In the blood plasma of humans or mammals, it is limited to local application, which may infect blood-borne diseases of humans or animals; collagen, as a foreign body, may cause wound fever and allergies; gelatin sponge has poor mechanical properties and is easy to break. There are also some hemostatic materials imported from abroad, such as instant gauze, which have a good hemostatic effect but are expensive, which limits their application range. Therefore, it is very important to have a hemostatic material with good hemostatic effect, strong antibacterial property, good ability to promote wound healing and degradable in vivo.

At present, chitosan as the main component has extensive research and application prospects as a hemostatic material. High biological activity, non-toxic, non-irritating, and can be degraded naturally. Many researchers choose chitosan to prepare porous hemostatic materials, and generally use a lower concentration of acetic acid solution to dissolve chitosan and obtain it by freeze-drying. However, the solubility of chitosan is relatively low, only 3% to 4%. The hemostatic material with lower chitosan concentration affects the hemostatic effect of the hemostatic material on the one hand, and affects the mechanical properties of the hemostatic material on the other hand. In addition, acetic acid is more irritating to the human body, and its use as a dissolving agent increases the unsafe use of materials. In order to improve the mechanical properties and biosafety performance of materials, it is necessary to select suitable natural biomaterials for blending and select non-toxic and harmless cross-linking agents for cross-linking, and at the same time, it is necessary to reduce or eliminate the content of acetic acid in the materials.

As an effective porous hemostatic material, it is not enough to select a suitable material, and it is also necessary to prepare the material with a specific pore structure and strong water absorption function. The general pore-making methods include particle dissolution method, phase separation method, freeze-drying method, gas foaming method, sintered microsphere method, etc., but how to choose a suitable pore-making method to prepare a pore structure with uniform pore size and high porosity , and have very good hydrophilicity and certain strength material, it is still a technical problem.

Contents of the invention

The technical problem to be solved by the present invention is to provide a chitosan product with simple preparation, high content of chitosan, natural degradation, fast hemostasis, good antibacterial effect, non-toxic, non-irritating, good mechanical properties and biological safety performance, and uniform pore size. 1. A preparation method of chitosan porous hemostatic sponge with high porosity.

The technical scheme that the present invention solves the problems of the technologies described above is as follows: a kind of preparation method of chitosan porous hemostatic sponge, comprises the following steps:

Step 1, the preparation of solution:

1) To prepare a chitosan solution, weigh ascorbic acid and add it to distilled water, fully stir and dissolve it in a water bath at 40°C-50°C to obtain a 0.05mol/L-0.3mol/L ascorbic acid solution; measure acetic acid and add it to the capacity Constant volume in the bottle to prepare an acetic acid solution with a concentration of 0.1% to 0.5% by volume; weigh chitosan and add it to the ascorbic acid solution, after mixing thoroughly, add the acetic acid solution, Stir evenly under water bath conditions to obtain a chitosan solution with a mass percentage concentration of 5% to 8%, which is stored for use, wherein the added acetic acid solution accounts for 0.2% to 0.5% of the total solution;

2) preparing a gelatin solution, weighing the gelatin and adding it to distilled water, fully dissolving it in a water bath at 40° C. to 85° C. to prepare a gelatin solution with a concentration of 0.5% to 4% by mass, and storing it for use;

3) preparing a glycerin solution, weighing glycerin and adding it to distilled water, stirring evenly to obtain a glycerin solution with a mass percentage concentration of 40% to 50%, and storing it for use;

4) preparing a calcium chloride solution, weighing the calcium chloride and adding it to distilled water to obtain a calcium chloride solution with a mass percentage concentration of 10% to 30%, which is stored for use;

5) Prepare a crosslinking agent, weigh tannic acid and add it to distilled water to prepare a tannic acid solution with a concentration of 0.5% to 5% by mass, and store it for use;

Step 2, the preparation of chitosan porous hemostatic sponge:

Take the chitosan solution, gelatin solution and glycerin solution according to the volume ratio of 3:3:1 and fully stir and mix them in a water bath at 25°C to 50°C to obtain a mixed milky white gel, and then take the calcium chloride solution Add, fully stir for 30min to 120min in a water bath at 25°C to 50°C, then add the tannic acid solution, and continue to stir for 20min to 50min in a water bath at 25°C to 50°C to obtain a mixed red gel solution. In the obtained mixed red gel solution, the calcium chloride solution and the tannic acid solution account for 0.5% to 4% and 0.01% to 0.1% respectively; take the mixed red gel solution and pour it into In the mould, it is pre-frozen for more than 12 hours to form an ice cube, and then vacuum freeze-dried for 48 hours to 72 hours to obtain a chitosan porous hemostatic sponge.

The invention has the beneficial effects of simple preparation, high content of chitosan in the finished product, natural degradation, fast hemostasis, good antibacterial effect, non-toxic, non-irritating, good mechanical properties and safety performance, uniform pore size (pore size is 100 μm～ 200μm), high porosity.

Preferably, the volumetric flask described in step one is a 100ml volumetric flask, which is convenient for a small amount of said acetic acid solution to be constant to volume.

Preferably, the preservation and use temperature environment of the chitosan solution, glycerin solution, calcium chloride solution and tannic acid solution in step 1 are all 30°C to 40°C to ensure that the chitosan solution, glycerin solution, chloride The molecular structure stability of calcium solution and tannic acid solution is beneficial to the preparation of chitosan porous hemostatic sponge.

Preferably, the storage and use temperature environment of the gelatin solution in step 1 is 30° C. to 50° C. to ensure the stability of the molecular structure of the gelatin solution and facilitate the preparation of chitosan porous hemostatic sponge.

Preferably, the pre-freezing temperature environment of the mixed red gel liquid in step 2 is -20°C to -80°C to ensure that the mixed red gel liquid is frozen in the form of ice cubes, which is beneficial to obtain chitosan porous hemostasis sponge.

detailed description

The principles and features of the present invention are described below, and the examples given are only used to explain the present invention, and are not intended to limit the scope of the present invention.

Embodiment 1, a kind of preparation method of chitosan porous hemostatic sponge, comprises the following steps:

Step 1, the preparation of solution:

1) To prepare a chitosan solution, weigh ascorbic acid and add it to distilled water, fully stir and dissolve it in a water bath at 40°C-50°C to obtain a 0.05mol/L-0.3mol/L ascorbic acid solution; measure acetic acid and add it to 100ml Constant volume in a volumetric flask to prepare an acetic acid solution with a concentration of 0.1% to 0.5% by volume; weigh chitosan and add it to the ascorbic acid solution, after mixing thoroughly, add the acetic acid solution, and place in a water bath at 40°C to 50°C Stir evenly to obtain a chitosan solution with a mass percent concentration of 5% to 8%, which is stored for use, wherein the added acetic acid solution accounts for 0.2% to 0.5% of the total solution;

2) Prepare a gelatin solution, weigh the gelatin and add it to distilled water, fully dissolve it in a water bath at 40°C to 85°C to prepare a 0.5% to 4% gelatin solution, store it at 30°C to 50°C for use;

3) Prepare a glycerin solution, weigh the glycerin and add it to distilled water, stir evenly to obtain a 40%-50% glycerin solution, store it at 30%-40°C for use;

4) preparing a calcium chloride solution, weighing the calcium chloride and adding it to distilled water to obtain a 10% to 30% calcium chloride solution, which is stored at 30% to 40°C for use;

5) Prepare a cross-linking agent, weigh tannic acid and add it to distilled water to prepare a 0.5% to 5% tannic acid solution, and store it at 30% to 40°C for use;

Step 2, the preparation of chitosan porous hemostatic sponge:

Get described chitosan solution, gelatin solution and glycerin solution according to the volume ratio of 3: 3: 1 and fully stir and mix in 25 ℃ of water bath conditions, obtain mixed milky white gel liquid, then get described calcium chloride solution and add, in Fully stir for 60 minutes in a water bath at 25°C, then add the tannic acid solution, and continue stirring for 50 minutes in a water bath at 25°C to obtain a mixed red gel. The calcium chloride solution and the tannic acid solution accounted for 0.5% and 0.01% respectively; the mixed red gel solution was taken, poured into a mold, and pre-frozen at -20°C for 12 hours to form an ice cube, and then Chitosan porous hemostatic sponge was obtained after vacuum freeze-drying for 48 hours.

Embodiment 2, a kind of preparation method of chitosan porous hemostatic sponge, comprises the following steps:

Step 1 is consistent with embodiment 1;

Step 2, the preparation of chitosan porous hemostatic sponge:

Get described chitosan solution, gelatin solution and glycerin solution according to the volume ratio of 3: 3: 1 and fully stir and mix in 40 ℃ of water bath conditions, obtain mixed milky white gel liquid, then get described calcium chloride solution and add, in Fully stir for 50 minutes in a water bath at 40°C, then add the tannic acid solution, and continue stirring for 40 minutes in a water bath at 40°C to obtain a mixed red gel. The calcium chloride solution and the tannic acid solution account for 2% and 0.05% respectively; the mixed red gel solution is taken, poured into a mold, and pre-frozen at -50°C for 12 hours to form an ice cube, and then Chitosan porous hemostatic sponge was obtained after vacuum freeze-drying for 60 hours.

Embodiment 3,, a kind of preparation method of chitosan porous hemostatic sponge, comprises the following steps:

Step 1 is consistent with embodiment 1;

Step 2, the preparation of chitosan porous hemostatic sponge:

Get described chitosan solution, gelatin solution and glycerol solution according to the volume ratio of 3: 3: 1 and fully stir and mix in 50 ℃ of water bath conditions, obtain mixed milky white gel liquid, then get described calcium chloride solution and add, in Fully stir for 30 minutes in a water bath at 50°C, then add the tannic acid solution, and continue stirring for 20 minutes in a water bath at 50°C to obtain a mixed red gel. The calcium chloride solution and the tannic acid solution accounted for 0.4% and 0.1% respectively; the mixed red gel solution was taken, poured into a mold, and pre-frozen at -80°C for 12 hours to form an ice cube, and then Chitosan porous hemostatic sponge was obtained after vacuum freeze-drying for 72 hours.

Experimental case 1:

Carry out ear artery hemostatic experiment with the porous chitosan hemostatic sponge obtained in Example 1: the porous chitosan hemostatic sponge is cut into 30 * 30 * 5mm specification for standby, take New Zealand experimental white rabbit, anesthetize, fix on the operating table, Shave off the hair on the rabbit's ear artery, take the left or right ear, cut off the ear artery 7cm from the ear tip, wipe off the floating blood, gently press it with a porous chitosan hemostatic sponge, and then take a 200g weight to press on the hemostatic sponge On the top, start timing, and choose the gelatin sponge sold in the market as a comparison. Compared with the gelatin sponge sold in the market, the hemostasis time is shortened by nearly double.

Experimental case 2:

Liver hemostasis experiment with the porous chitosan hemostatic sponge prepared in Example 1: cut the porous chitosan hemostatic sponge into a size of 30×30×5mm for subsequent use, take the New Zealand experimental blue rabbit, anesthetize it, and fix it on the operating table. Shave off the hair on the abdomen of the rabbit, cut a wound about 4 cm long in the upper part of the abdomen, take out the liver, cut a wound 1 cm deep and 1 cm long on the liver, wipe off the floating blood on the surface of the liver, and use porous chitosan to stop the bleeding Press the wound with a sponge, put a 100g weight on the hemostatic sea surface, and start timing. Compared with the gelatin sponge sold in the market, the hemostasis time is shortened by nearly double.

The above descriptions are only preferred embodiments of the present invention, and are not intended to limit the present invention. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principles of the present invention shall be included in the protection of the present invention. within range.

Claims (5)

1. a preparation method of chitosan porous hemostatic sponge, is characterized in that, comprises the following steps: Step 1, the preparation of solution: 1) To prepare a chitosan solution, weigh ascorbic acid and add it to distilled water, fully stir and dissolve it in a water bath at 40°C-50°C to obtain a 0.05mol/L-0.3mol/L ascorbic acid solution; measure acetic acid and add it to the capacity Constant volume in the bottle to prepare an acetic acid solution with a concentration of 0.1% to 0.5% by volume; weigh chitosan and add it to the ascorbic acid solution, after mixing thoroughly, add the acetic acid solution, Stir evenly under water bath conditions to obtain a chitosan solution with a mass percentage concentration of 5% to 8%, which is stored for use, wherein the added acetic acid solution accounts for 0.2% to 0.5% of the total solution; 2) Prepare a gelatin solution, weigh the gelatin and add it to distilled water, fully dissolve it in a water bath at 40°C to 85°C to prepare a gelatin solution with a concentration of 0.5% to 4% by mass, and store it at 30°C to 50°C for use ; 3) preparing a glycerin solution, weighing glycerin and adding it to distilled water, stirring evenly to obtain a glycerin solution with a mass percentage concentration of 40% to 50%, and storing it for use; 4) preparing a calcium chloride solution, weighing the calcium chloride and adding it to distilled water to obtain a calcium chloride solution with a mass percentage concentration of 10% to 30%, which is stored for use; 5) Prepare a crosslinking agent, weigh tannic acid and add it to distilled water to prepare a tannic acid solution with a concentration of 0.5% to 5% by mass, and store it for use; Step 2, the preparation of chitosan porous hemostatic sponge: Take the chitosan solution, gelatin solution and glycerin solution according to the volume ratio of 3:3:1 and fully stir and mix them in a water bath at 25°C to 50°C to obtain a mixed milky white gel, and then take the calcium chloride solution Add, fully stir for 30 minutes to 120 minutes in a water bath at 25°C to 50°C, then add the tannic acid solution, and continue to stir for 20 minutes to 50 minutes in a water bath at 25°C to 50°C to obtain a mixed red gel solution. In the obtained mixed red gel solution, the calcium chloride solution and the tannic acid solution account for 0.5% to 4% and 0.01% to 0.1% respectively; take the mixed red gel solution and pour it into In the mould, it is pre-frozen for more than 12 hours to form an ice cube, and then vacuum freeze-dried for 48 hours to 72 hours to obtain a chitosan porous hemostatic sponge. 2. the preparation method of a kind of chitosan porous hemostatic sponge according to claim 1 is characterized in that, the volumetric flask described in step one is a volumetric flask of 100ml. 3. the preparation method of a kind of chitosan porous hemostatic sponge according to claim 1 and 2 is characterized in that, the preservation of chitosan solution, glycerin solution, calcium chloride solution and tannic acid solution described in step one The operating temperature environment is 30 ℃ ~ 40 ℃. 4. The preparation method of a chitosan porous hemostatic sponge according to claim 1 or 2, characterized in that the preservation and use temperature environment of the gelatin solution described in step 1 is 30° C. to 50° C. 5. the preparation method of a kind of chitosan porous hemostatic sponge according to claim 1 or 2 is characterized in that, the prefreezing temperature environment of the mixed red gel liquid described in step 2 is -20 ℃～-80 ℃ .