Hemostatic powder dressing and preparation method thereof

Abstract

The invention relates to a hemostatic powder dressing and discloses a hemostatic powder dressing and a preparation method thereof.The preparation method comprises the steps that chitosan powder is dissolved in an acetic acid solution to obtain an acetic acid solution, the acetic acid solution is homogenized, and then solid-liquid separation, cleaning and freeze-drying treatment are conducted in sequence to obtain chitosan sponge; and grinding the chitosan sponge, mixing the ground chitosan sponge with the bioglass, and sieving to obtain the hemostatic powder dressing. Chitosan and bioglass are combined by means of chemical reaction and physical mixing, absorption of wound exudate is effectively promoted while a physical barrier is formed on the surface of a wound, finally, the hemostasis process is accelerated, and the hemostasis effect of chitosan is effectively optimized.

Description

Technical Field

[0001] The invention relates to a powder dressing, in particular to a hemostatic powder dressing. Background Art

[0002] At present, the commonly used hemostatic materials in clinical practice include hemostatic gauze, hemostatic fibers, hemostatic bandages, etc. However, they have problems such as long hemostasis time, easy adhesion to the wound, and inability to prevent wound infection and suppuration during use.

[0003] Hemostatic materials are mainly divided into two categories: biological and chemical. Biological hemostatic materials mainly include fibrin glue and collagen, while chemical hemostatic materials mainly include chitosan, microporous inorganic materials, soluble hemostatic gauze, tissue glue, etc. The main problems of fibrin glue and collagen are high cost, slow hemostasis speed and the risk of disease infection; porous zeolite may have the problem of exothermic burns on surface tissues during use; soluble hemostatic gauze has problems such as long hemostasis time and poor hemostatic performance for patients with rapid bleeding.

[0004] Chitosan is a new type of hemostatic material developed in recent years. It is gradually used in clinical practice due to its good biocompatibility with the human body, biodegradability, and ability to promote tissue regeneration. As early as 1983, Malette WG et al. found that chitosan has good hemostatic function. Chitosan has high adhesion and adsorption. It can interact with components such as red blood cells, platelets and plasma proteins in the blood to form a layer of thrombus structure adhering to the wound, effectively sealing the wound and preventing further bleeding. At the same time, chitosan can act as an activator of coagulation reaction, promote the activation and aggregation of coagulation factors, bind to specific components in the blood, and accelerate the coagulation process. Chitosan can also promote the aggregation and adhesion of platelets and red blood cells by interacting with molecules such as fibrinogen and fibronectin in plasma, thereby forming a stable hemostatic thrombus on the wound and accelerating hemostasis.

[0005] Although the prior art has shown that chitosan has a good hemostatic effect, its specific application and optimization of the hemostatic effect (such as further accelerating wound hemostasis) are still problems that need to be solved in the art. Summary of the invention

[0006] Aiming at the problem that the hemostatic effect of chitosan in the prior art still needs to be further optimized, the present invention provides a hemostatic powder dressing and a preparation method thereof.

[0007] In order to solve the above technical problems, the present invention is solved by the following technical solutions: A method for preparing a hemostatic powder dressing comprises the steps of dissolving chitosan powder in an acetic acid solution to obtain an acetic acid solution, homogenizing the acetic acid solution, sequentially performing solid-liquid separation, washing and freeze-drying to obtain a chitosan sponge, and then grinding the chitosan sponge and mixing it with bioglass and sieving it to obtain a hemostatic powder dressing.

[0008] As a preference, the method specifically comprises the following steps: Step S1, weighing chitosan powder, and dissolving it in acetic acid solution to obtain acetic acid solution; Step S2, homogenizing the acetic acid solution to obtain a homogenized chitosan solution; Step S3, dissolving NaHCO3 in 10×PBS buffer to obtain NaHCO3 / 10×PBS buffer; Step S4, slowly adding the homogenized chitosan solution into NaHCO3 / 10×PBS buffer for reaction; Step S5, leaving the mixed solution after the reaction in step S4 to stand at room temperature to separate the solid from the liquid; Step S6, recovering the solid in the reaction system, soaking and washing, and freeze-drying the obtained solid to obtain a chitosan sponge; Step S7, grinding the freeze-dried chitosan sponge into powder; Step S8, mixing the ground powder with bioglass at a mass ratio of 8-9:1 to obtain chitosan powder dressing.

[0009] Preferably, the chitosan powder is natural chitosan powder, and dissolving the chitosan powder in the acetic acid solution is dissolving the natural chitosan powder in the acetic acid solution to obtain an acetic acid solution with a concentration of 1.5-3% (w / v).

[0010] Preferably, the chitosan powder comprises natural chitosan powder and modified chitosan powder, and the chitosan powder is dissolved in acetic acid solution by dissolving the natural chitosan powder and the modified chitosan powder in acetic acid solution respectively to obtain 1.5-3% (w / v) natural chitosan solution and 1.5-3% (w / v) modified chitosan solution respectively, and then the natural chitosan solution and the modified chitosan solution are mixed to obtain an acetic acid solution with a concentration of 1.5-3% (w / v), wherein the modified chitosan powder comprises carboxymethyl chitosan and / or quaternary ammonium chitosan.

[0011] Preferably, the homogenization temperature during the homogenization of the acetic acid solution is 4-25° C., and the homogenization time is 24-240 h.

[0012] Preferably, in step S5, the mixed solution is allowed to stand at room temperature for 10-30 minutes.

[0013] Preferably, the soaking and cleaning in step S6 comprises the following steps: Step S61, soaking and washing with 1×PBS for 10 min; Step S62, discard PBS, and then soak and wash with deionized water twice, each time for 10 min; Step S63, discard the deionized water and obtain the solid matter.

[0014] Preferably, freeze-drying the solid obtained in step S6 comprises pre-freezing the solid and then transferring it to -20°C and freezing it for 30 minutes.

[0015] Preferably, in step S7, the grinding speed is 600-900 rpm, and the grinding time is 5-10 min.

[0016] A hemostatic powder dressing is prepared by using the above-mentioned method for preparing a hemostatic powder dressing.

[0017] The present invention has significant technical effects due to the adoption of the above technical solution: By combining chitosan and bioglass through chemical reaction and physical mixing, a physical barrier is formed on the wound surface while effectively promoting the absorption of wound exudate, ultimately accelerating the hemostasis process and effectively optimizing the hemostatic effect of chitosan. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 It is a scanning electron microscope picture of the hemostatic powder of the present invention.

[0019] Figure 2 A diagram showing the water absorption effect of the blood powder of the present invention.

[0020] Figure 3 A diagram showing the hemostatic effect of the hemostatic powder of the present invention.

[0021] Figure 4 The toxicity test of the hemostatic powder of the present invention on cells. DETAILED DESCRIPTION

[0022] The present invention is further described in detail below in conjunction with the accompanying drawings and embodiments.

[0023] Example 1 like Figure 1 As shown, a method for preparing a hemostatic powder dressing is provided, wherein chitosan powder is dissolved in an acetic acid solution to obtain an acetic acid solution, the acetic acid solution is homogenized and sequentially subjected to solid-liquid separation, washing and freeze-drying treatment to obtain a chitosan sponge, and then the chitosan sponge is ground and mixed with bioglass and sieved to obtain a hemostatic powder dressing.

[0024] The specific steps include: Step S1, weigh chitosan powder, in this embodiment, the chitosan powder is natural chitosan powder, dissolve it in 1.25% (w / v) acetic acid solution to a final concentration of 2.5% (w / v) acetic acid solution, and stir at room temperature overnight; Step S2, homogenizing and balancing the dissolved acetic acid solution at 4° C. for 48 hours to obtain a homogenized chitosan solution; Step S3, dissolving NaHCO3 in 10×PBS to a final concentration of 0.45 M NaHCO3 / 10×PBS buffer; Step S4, slowly adding the homogenized chitosan solution into NaHCO3 / 10×PBS at a ratio of 7:10 for reaction, the reaction speed is 400-1000 rpm, and the reaction speed in this embodiment is specifically 700 rpm; Step S5, leaving the reaction mixture to stand at room temperature for 30 min to separate the solid and the liquid; Step S6, recovering the solid in the reaction system, and soaking and washing it with 1×PBS for 10 min; discarding the PBS, and then soaking and washing it with deionized water twice, each time for 10 min; discarding the deionized water, transferring the obtained solid to a -20°C refrigerator and freezing it for about 30 min, and then freeze-drying it to obtain a freeze-dried sponge; Step S7, grinding the freeze-dried sponge into powder using a ball mill; Step S8, the powder and bioglass are mixed evenly in a ratio of 9:1, and then passed through a 100 μm sieve to obtain a chitosan powder dressing.

[0025] Bioglass has good biocompatibility, high bioactivity, biodegradability and bone repair properties, and its chemical composition is very similar to the natural inorganic elements found in the human body, ensuring safety and stability. Bioglass also has good ion loading capacity and multifunctional biological properties such as antibacterial activity, which can promote wound healing while reducing the risk of infection.

[0026] like Figure 1 As shown, in this embodiment, the chitosan powder prepared was photographed by scanning electron microscopy, and it was determined that the hemostatic powder had obvious pores and the particle size was less than 100 μm; such a structure helps the hemostatic powder to fit the wound perfectly, thereby better exerting its function of absorbing exudate and accelerating hemostasis.

[0027] Example 2 The same as Example 1, except that the chitosan powder in step S1 of this embodiment includes natural chitosan powder and modified chitosan powder. In this embodiment, the modified chitosan powder is carboxymethyl chitosan. The natural chitosan powder and the carboxymethyl chitosan powder were dissolved in acetic acid solution to obtain 2.5% (w / v) natural chitosan solution and 2.5% (w / v) carboxymethyl chitosan solution, respectively. Then, the natural chitosan solution and the carboxymethyl chitosan solution were evenly mixed in a ratio of 9:1.

[0028] Carboxymethyl chitosan is an important water-soluble chitosan derivative with many medical effects, such as promoting wound healing, hemostasis, scar inhibition, analgesia and antibacterial effects. It is also widely used in chemical industry, environmental protection and health care products. Carboxymethyl chitosan has good biocompatibility and biodegradability, and is widely used in hydrogels and wound healing biomaterials. It can quickly stop bleeding, and the hemostatic mechanism does not rely on low molecular weight heparin.

[0029] Therefore, its combined use with chitosan can improve the water solubility of the product while accelerating the wound hemostasis process. That is, in this embodiment, by combining chitosan, carboxymethyl chitosan and bioglass through chemical reaction and physical mixing, the hemostatic effect of the prepared hemostatic powder is further enhanced, and the hemostatic effect of chitosan is further optimized.

[0030] Example 3 The same as Example 2, except that in step S1 of this example, the natural chitosan solution and the carboxymethyl chitosan solution are evenly mixed in a ratio of 4:1.

[0031] Example 4 The same as Example 2, except that in step S1 of this example, the natural chitosan solution and the carboxymethyl chitosan solution are evenly mixed in a ratio of 19:1.

[0032] Example 5 This embodiment provides a hemostatic powder for use as a dressing, which is prepared by Example 1 and Examples 2-4, weighed after grinding, and an equal volume of simulated body fluid is added, and the powder is soaked at 37° C. for 1 hour to absorb water and then weighed again.

[0033] Table 1 Effect of adding different contents of carboxymethyl chitosan on the absorption of exudate by hemostatic powder

[0034] It was found that the carboxyl content would affect the hemostatic material's ability to absorb exudate. As the carboxyl content increased, the ability to absorb exudate would slightly decrease, but it could still absorb about 10 times its own weight of liquid. Figure 2 shown.

[0035] Example 6 Hemostasis Example Twenty healthy adult SD rats weighing 180±10g were selected. After successful anesthesia, the SD rats were fixed on the dissecting table and disinfected with iodine solution. A longitudinal incision of about 2cm in length was made along the midline of the abdomen under the costal arch, and the depth was first to the full layer of skin. Then the abdominal muscles were lifted and cut along the midline of the abdomen, paying attention to avoid important organs in the abdomen. The abdomen was gently squeezed, and the middle lobe of the liver was squeezed out from the incision. Care was taken to avoid damaging the middle lobe of the liver. The peritoneal fluid around the liver was absorbed with sterile gauze, and the sterile gauze weighed on a balance was placed under the middle lobe of the liver. The middle lobe of the liver was quickly removed, and the tangent line was about 2cm above the lower edge of the middle lobe of the liver.

[0036] At the same time, the hemostatic powder dressing in Example 5 was immediately applied, and the time was counted until the bleeding stopped, and the bleeding time was recorded. Figure 3 shown.

[0037] Experiments have shown that the hemostatic powder dressing exhibits excellent therapeutic effects in hemostatic repair in rats, and also provides a new option for rapid hemostasis of injuries in clinical patients.

[0038] Example 7 Cytotoxicity Test like Figure 4 As shown, the hemostatic powder prepared in Example 5 was subjected to a cytotoxicity test. Cell culture medium was added to the hemostatic powder, and after soaking for 24 hours, the supernatant was taken to obtain an extract.

[0039] Human mesenchymal stem cells in the logarithmic growth phase were obtained. When the confluence reached 80-90%, they were digested, centrifuged, and resuspended to make a cell suspension, which was inoculated into a 96-well plate with 5 replicate wells. After the cells adhered to the wall, the extract was added to culture the cells, and the cell proliferation was detected on Day 1, Day 3, and Day 5 after extraction.

[0040] It is easy to understand that those skilled in the art can combine, split, reorganize, etc. the embodiments of the present application to obtain other embodiments based on one or several embodiments provided in the present application, and these embodiments do not exceed the protection scope of the present application.

[0041] In conclusion, the above is only a preferred embodiment of the present invention, and all equivalent changes and modifications made according to the scope of the patent application of the present invention should fall within the scope of the patent of the present invention.

Claims

1. A method for preparing a hemostatic powder dressing, characterized in that: The acetic acid solution is obtained by dissolving chitosan powder in an acetic acid solution, the acetic acid solution is homogenized, and then solid-liquid separation, washing and freeze-drying are performed in sequence to obtain a chitosan sponge, and then the chitosan sponge is ground and mixed with bioglass and sieved to obtain a hemostatic powder dressing.

2. The method for preparing a hemostatic powder dressing according to claim 1, characterized in that: The specific steps include: Step S1, weighing chitosan powder, and dissolving it in acetic acid solution to obtain acetic acid solution; Step S2, homogenizing the acetic acid solution to obtain a homogenized chitosan solution; Step S3, dissolving NaHCO3 in 10×PBS buffer to obtain NaHCO3 / 10×PBS buffer; Step S4, slowly adding the homogenized chitosan solution into NaHCO3 / 10×PBS buffer for reaction; Step S5, leaving the mixed solution after the reaction in step S4 to stand at room temperature to separate the solid from the liquid; Step S6, recovering the solid in the reaction system, soaking and washing, and freeze-drying the obtained solid to obtain a chitosan sponge; Step S7, grinding the freeze-dried chitosan sponge into powder; Step S8, mixing the ground powder with bioglass at a mass ratio of 8-9:1 to obtain chitosan powder dressing.

3. The method for preparing a hemostatic powder dressing according to claim 2, characterized in that: The chitosan powder is natural chitosan powder, and dissolving the chitosan powder in the acetic acid solution is dissolving the natural chitosan powder in the acetic acid solution to obtain an acetic acid solution with a concentration of 1.5-3% (w / v).

4. The method for preparing a hemostatic powder dressing according to claim 2, characterized in that: The chitosan powder includes natural chitosan powder and modified chitosan powder. The chitosan powder is dissolved in acetic acid solution by dissolving the natural chitosan powder and the modified chitosan powder in acetic acid solution respectively to obtain 1.5-3% (w / v) natural chitosan solution and 1.5-3% (w / v) modified chitosan solution respectively, and then the natural chitosan solution and the modified chitosan solution are mixed to obtain an acetic acid solution with a concentration of 1.5-3% (w / v).

5. A method for preparing a hemostatic powder dressing according to claim 3 or 4, characterized in that: The homogenization temperature during the homogenization of the acetic acid solution is 4-25° C., and the homogenization time is 24-240 h.

6. A method for preparing a hemostatic powder dressing according to claim 3 or 4, characterized in that: In step S5, the mixed solution is allowed to stand at room temperature for 10-30 minutes.

7. A method for preparing a hemostatic powder dressing according to claim 3 or 4, characterized in that: The soaking and cleaning in step S6 includes the following steps: Step S61, soaking and washing with 1×PBS for 10 min; Step S62, discard PBS, and then soak and wash with deionized water twice, each time for 10 min; Step S63, discard the deionized water and obtain the solid matter.

8. The method for preparing a hemostatic powder dressing according to claim 7, characterized in that: In step S6, freeze-drying the obtained solid matter specifically comprises pre-freezing the solid matter first, and then transferring it to -20°C and freezing it for 30 minutes.

9. A method for preparing a hemostatic powder dressing according to claim 3 or 4, characterized in that: In step S7, the grinding speed is 600-900 rpm, and the grinding time is 5-10 min.

10. A hemostatic powder dressing, characterized in that: The hemostatic powder dressing is prepared by the method for preparing the hemostatic powder dressing according to any one of claims 1 to 9.

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