A wound dressing with photothermal antibacterial, hemocoagulant, and easy-to-remove properties, its preparation method, and its application.
Wound dressings prepared by cross-linking hemoglobin and sericin solve the problems of rapid hemostasis and antibacterial properties, achieving easy peeling and photothermal antibacterial effects, and are suitable for rapid healing of accidental wounds.
Patent Information
- Application Number
- CN202411711830.5
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- Filing Date
- 2024-11-27
- Publication Date
- 2026-01-30
- Estimated Expiration
- 2044-11-27
AI Technical Summary
Existing wound dressings are inadequate for achieving rapid hemostasis and antibacterial properties in accidental injuries, and may lead to bacterial infection and secondary injury during the healing process.
Wound dressings are prepared by combining hemoglobin and sericin. A dense fibrin network is formed through cross-linking reaction, which promotes blood clotting. The photothermal effect of hemoglobin is used to achieve antibacterial properties. The dressing is easy to peel off from the wound site.
This wound dressing achieves rapid hemostasis, antibacterial properties, and easy removal. It has good biocompatibility, photothermal antibacterial properties, and can effectively kill bacteria and promote the formation of fibrin networks, reducing secondary damage.
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Figure CN119455082B_ABST
Abstract
Description
TECHNICAL FIELD
[0001] The present application relates to the technical field of biological materials, in particular to a wound dressing with photothermal antibacterial hemostatic and easy peeling and a preparation method and application thereof. BACKGROUND
[0002] Uncontrollable massive bleeding in deep tissue in accidental trauma can lead to high mortality. In addition, due to the complex environment in which accidental trauma is usually exposed, the wound is susceptible to bacterial infection, which further delays wound healing. Therefore, an ideal wound dressing needs to have antibacterial activity and rapid hemostatic properties. Sponge hemostatic has high liquid absorption due to its porous structure, and exhibits excellent hemostatic effect in the hemostasis of uncontrollable massive bleeding in deep tissue, and is considered to be a potential candidate for pre-hospital treatment.
[0003] In recent years, hemostatic sponges prepared by compounding various components have been studied to achieve efficient hemostasis and antibacterial activity. Natural proteins have good biocompatibility and degradation performance, and their degradation products are not only non-toxic but also suitable for human use. In particular, natural proteins with high abundance, availability and low cost are ideal raw materials for preparing biomaterials with practical application value.
[0004] Hemoglobin is a physiological oxygen transport metal protein present in red blood cells. At present, the application of hemoglobin in biological medical materials mainly focuses on photothermal antibacterial and chronic skin wound repair. Its application in hemostasis has not been widely developed. SUMMARY
[0005] In view of the deficiencies in the above background art, the present application provides a wound dressing with photothermal antibacterial hemostatic and easy peeling and a preparation method and application thereof. The protein wound dressing prepared by the method is composed of only hemoglobin and silk glue, and has good biocompatibility. The composite protein dressing has hemostatic properties that single-component does not have. By promoting the aggregation of red blood cells and the formation of fibrin film, rapid coagulation is achieved. The fibrin film formed on the surface of the dressing during hemostasis also isolates the dressing from the injured tissue, so that the dressing is easy to peel off from the injured site, avoiding secondary damage. In addition, it also has photothermal antibacterial activity.
[0006] The first object of the present application is to provide a preparation method of a wound dressing with photothermal antibacterial hemostatic and easy peeling, comprising the following steps:
[0007] Dissolve silk glue in water, then add hemoglobin and carbodiimide in sequence and stir uniformly, then add N-hydroxysuccinimide and stir to perform cross-linking reaction, to obtain a cross-linked solution;
[0008] The activated proteins in the cross-linking solution are aggregated and cross-linking reaction occurs by applying external force to the cross-linking solution, and the hemoglobin sponge precursor is collected by aggregating the aggregates;
[0009] The hemoglobin sponge precursor is pre-cooled and completely freeze-dried to obtain the sericin hemoglobin sponge;
[0010] The sericin hemoglobin sponge is dialyzed in water for 2-3 days, pre-cooled again and freeze-dried to obtain the hemoglobin sericin dressing.
[0011] Preferably, the mass ratio of the sericin protein, the carbodiimide and the N-hydroxysuccinimide is 5:1-10:1-5;
[0012] The mass ratio of the sericin protein and the hemoglobin is 10:1-5;
[0013] The cross-linking reaction temperature is 0-50℃, and the reaction time is greater than 2 h.
[0014] Preferably, the way of applying external force includes centrifugation or pressurization.
[0015] Preferably, when the external force is applied to the cross-linking solution, centrifugation is used to aggregate the proteins in the cross-linking solution and make cross-linking reaction occur; wherein the centrifugal speed is 1000-10000 rpm, and the centrifugal time is 1-20 min.
[0016] Preferably, the pre-cooling temperature is-80--20℃, and the pre-cooling time is 12-24 h; the freeze-drying time is 10-24 h.
[0017] Preferably, the molecular weight of the sericin protein and the hemoglobin is ≥8000 MW.
[0018] Preferably, during the dialysis of the sericin hemoglobin sponge in water, the dialysis bag with a molecular weight cut-off of 3500 MW is used.
[0019] Preferably, when the sericin protein is dissolved in water, it is heated to complete dissolution by a boiling water bath, and then cooled to room temperature.
[0020] The second object of the present application is to provide a wound dressing with photothermal antibacterial hemostatic and easy peeling.
[0021] The third object of the present application is to provide the use of a wound dressing with photothermal antibacterial hemostatic and easy peeling in the preparation of antibacterial or hemostatic dressing.
[0022] Compared with the prior art, the present application has the following beneficial effects:
[0023] The application provides a light-thermal antibacterial hemostatic and easy-to-peel wound dressing and a preparation method and application thereof. The silk sericin hemoglobin wound dressing prepared by the application has good biocompatibility, light-thermal antibacterial activity, rapid hemostasis and easy-to-peel characteristics from a wound site. Hemoglobin has a near-infrared light-thermal effect, and can convert light energy into heat under the irradiation of 808 nm near-infrared light, so that bacteria are killed, and therefore the dressing has light-thermal antibacterial performance. In addition, the addition of hemoglobin makes the pore size of the original silk sericin wound dressing smaller, which is beneficial to the aggregation of red blood cells and platelets in the hemostasis process. Moreover, hemoglobin has the effect of promoting hemostasis, and when the composite protein dressing contacts blood, it can promote the formation of a dense fibrin network, thereby accelerating hemostasis. At the same time, the dense fibrin network on the surface of the dressing can isolate the hemostatic dressing from the tissue, so that the wound dressing is easy to peel off from the damaged site. The components of the dressing prepared by the application are only silk sericin protein and hemoglobin, and therefore the biocompatibility is good, and the application has great development prospects.
[0024] The application has high practical application value from the preparation method and performance. BRIEF DESCRIPTION OF DRAWINGS
[0025] Figure 1 Fig. 1 is a scanning electron microscope image of the silk sericin hemoglobin dressing obtained in Examples 1-3 and an element scanning result of Example 2.
[0026] Figure 2 Fig. 2 is a cyclic compression stress-strain curve of the silk sericin hemoglobin dressing obtained in Examples 1-3.
[0027] Figure 3 Fig. 3 is an antibacterial effect of the silk sericin hemoglobin dressing obtained in Examples 1-3 under the irradiation of 808 nm near-infrared light.
[0028] Figure 4 Fig. 4 is a scanning electron microscope image of the silk sericin hemoglobin dressing after interaction with red blood cells.
[0029] Figure 5 Fig. 5 is a scanning electron microscope image of the silk sericin hemoglobin dressing after interaction with blood plasma.
[0030] Figure 6 Fig. 6 is a statistical result of the hemostasis index of the silk sericin hemoglobin dressing obtained in Examples 1-3. DETAILED DESCRIPTION
[0031] In order to enable those skilled in the art to better understand the technical solutions of the application and implement the same, the application will be further described below in conjunction with specific examples and drawings, but the examples are not limiting on the application.
[0032] The application aims to provide a wound dressing with photothermal antibacterial hemostatic and easy peeling and a preparation method and application thereof.
[0033] To achieve the above-mentioned purpose, the application provides a preparation method of a wound dressing with photothermal antibacterial hemostatic and easy peeling, comprising the following steps:
[0034] The silk fibroin is dissolved in water, then the hemoglobin and carbodiimide are added in sequence and stirred uniformly, and then the N-hydroxysuccinimide is added and stirred to perform cross-linking reaction, so as to obtain a cross-linking solution;
[0035] An external force is applied to the cross-linking solution to make the activated proteins in the cross-linking solution aggregate and perform cross-linking reaction, and the aggregates are collected to obtain a hemoglobin sponge precursor;
[0036] The hemoglobin sponge precursor is pre-cooled and completely freeze-dried to obtain a silk fibroin hemoglobin sponge;
[0037] The silk fibroin hemoglobin sponge is dialyzed in water for 2-3 days, pre-cooled again and freeze-dried to obtain a hemoglobin silk fibroin dressing.
[0038] The preparation method provided by the application can construct a dressing with good porosity and good photothermal conversion efficiency under 808 nm irradiation, thereby achieving antibacterial effect.
[0039] The mass ratio of the silk fibroin, the carbodiimide and the N-hydroxysuccinimide is 5:1-10:1-5, and the preferable mass ratio is 5:5:3; the mass ratio of the silk fibroin to the hemoglobin is 10:1-5.
[0040] The cross-linking reaction temperature is 0-50℃, and the reaction time is greater than 2 h.
[0041] The external force application mode includes centrifugation or pressurization. Specifically, when the external force is applied to the cross-linking solution, centrifugation is adopted to make the proteins in the cross-linking solution aggregate and perform cross-linking reaction; wherein the centrifugation speed is 1000-10000 rpm, and the centrifugation time is 1-20 min.
[0042] The pre-cooling temperature is-80--20℃, the pre-cooling time is 12-24 h, and the freeze-drying time is 10-24 h.
[0043] The molecular weight of the sericin and hemoglobin is ≥8000 MW.
[0044] The molecular weight cut-off of the dialysis bag used in the dialysis of the sericin hemoglobin sponge in water is 3500 MW.
[0045] When the sericin is dissolved in water, it is heated to complete dissolution by a boiling water bath, and then cooled to room temperature.
[0046] In an embodiment, a method for preparing a sericin hemoglobin wound dressing comprises the following steps:
[0047] (1) Dissolve sericin in water and heat to complete dissolution, then add hemoglobin and carbodiimide in sequence and stir for 15 min, and finally add N-hydroxysuccinimide for cross-linking reaction to obtain a flocculent sericin and hemoglobin cross-linking reaction solution.
[0048] The molecular weight of the sericin and hemoglobin in step (1) is ≥8000 MW.
[0049] The mass ratio of sericin protein: carbodiimide: N-hydroxysuccinimide in step (1) is 5: 1-10: 1-5, and the mass ratio of sericin and hemoglobin is 10: 1-5.
[0050] The cross-linking reaction time in step (1) is ≥2 h, and the cross-linking reaction temperature is 0-50°C.
[0051] The process is to cross-link sericin and hemoglobin by using the carbodiimide protein coupling method.
[0052] (2) Centrifuge the cross-linking reaction solution obtained in step (1) to obtain a sericin and hemoglobin sponge precursor after removing the supernatant. The purpose of centrifugation is to increase the probability of collision and cross-linking of activated carboxyl and amino groups in sericin and hemoglobin.
[0053] (3) Pre-cool and completely freeze-dry the sericin and hemoglobin sponge precursor obtained in step (2) to obtain a sericin hemoglobin sponge;
[0054] (4) Dialyze the sericin hemoglobin sponge obtained in step (3) in water for 2-3 days, and then completely freeze-dry after pre-cooling again to obtain a wound dressing containing only sericin and hemoglobin.
[0055] The purpose of dialysis is to remove residual carbodiimide and N-hydroxysuccinimide in the product of step (3), and finally obtain a wound dressing containing only sericin and hemoglobin.
[0056] The centrifugation speed in step (2) is ≥1000 rpm, and the centrifugation time is ≥1 min.
[0057] The precooling temperature in the preparation process is 20℃ or higher, and the precooling time is 1 h or longer.
[0058] In the step (4), the molecular weight cut-off of the dialysis bag used is 3500 MW.
[0059] The water used in the preparation process is deionized water.
[0060] The second aspect of the present application provides a photothermal antibacterial hemostatic and easy-to-peel wound dressing.
[0061] The third aspect of the present application provides the use of a photothermal antibacterial hemostatic and easy-to-peel wound dressing in the preparation of an antibacterial or hemostatic agent.
[0062] It should be noted that the experimental methods used in the present application are conventional methods unless otherwise specified; the reagents and materials used are commercially available unless otherwise specified.
[0063] The silk fibroin (abbreviated as silk glue) and hemoglobin used in the following examples are from the laboratory. The silk glue is extracted by conventional high-temperature aqueous method. The hemoglobin is extracted by conventional water crushing extraction method.
[0064] Example 1
[0065] The photothermal antibacterial hemostatic and easy-to-peel wound dressing and its preparation method comprise the following steps:
[0066] Dissolve 500 mg of silk fibroin in 50 mL of deionized water and heat to complete dissolution using a boiling water bath, and then cool to room temperature. Then add 50 mg of hemoglobin and 500 mg of carbodiimide in turn and stir for 15 min, and finally add 300 mg of N-hydroxysuccinimide and stir at room temperature for 2 h or more. Centrifuge the crosslinking solution (6000 rpm, 15 min), and obtain the silk hemoglobin sponge precursor after removing the supernatant. The precursor is pre-cooled at -80℃ for 12 h and freeze-dried for 10 h or more to obtain a silk hemoglobin sponge with residual crosslinking agent. The sponge is dialyzed in deionized water using a 3500 MW dialysis bag for 2-3 days, and then pre-cooled at -80℃ for 12 h and freeze-dried for 10 h or more to obtain a silk hemoglobin sponge.
[0067] Experimental determination shows that the product has antibacterial activity, high compression strength, aggregated red blood cells and rapid blood clotting performance.
[0068] Example 2
[0069] The photothermal antibacterial hemostatic and easy-to-peel wound dressing and its preparation method comprise the following steps:
[0070] The silk sericin protein 500 mg was dissolved in 50 mL of deionized water and heated to complete dissolution, and then cooled to room temperature. Then 150 mg of hemoglobin and 500 mg of carbodiimide were added in turn and stirred for 15 min, and finally 300 mg of N-hydroxysuccinimide was added and stirred at room temperature for 2 h. The crosslinking solution was centrifuged (6000 rpm, 15 min), and the supernatant was removed to obtain a silk sericin hemoglobin sponge precursor. The precursor was pre-cooled at -80°C for 12 h and freeze-dried for more than 10 h to obtain a silk sericin hemoglobin sponge with residual crosslinking agent. The sponge was dialyzed in deionized water for 2-3 days using a 3500 MW dialysis bag, and then pre-cooled at -80°C for 12 h and freeze-dried for more than 10 h to obtain a silk sericin hemoglobin sponge.
[0071] Experimental determination showed that the product had antibacterial activity, high compression strength, aggregated red blood cells and rapid blood clotting properties.
[0072] Example 3
[0073] A photothermal antibacterial and hemostatic wound dressing with easy peeling and a preparation method thereof, comprising the following steps:
[0074] The silk sericin protein 500 mg was dissolved in 50 mL of deionized water and heated to complete dissolution, and then cooled to room temperature. Then 150 mg of hemoglobin and 500 mg of carbodiimide were added in turn and stirred for 15 min, and finally 300 mg of N-hydroxysuccinimide was added and stirred at room temperature for 2 h. The crosslinking solution was centrifuged (6000 rpm, 15 min), and the supernatant was removed to obtain a silk sericin hemoglobin sponge precursor. The precursor was pre-cooled at -80°C for 12 h and freeze-dried for more than 10 h to obtain a silk sericin hemoglobin sponge with residual crosslinking agent. The sponge was dialyzed in deionized water for 2-3 days using a 3500 MW dialysis bag, and then pre-cooled at -80°C for 12 h and freeze-dried for more than 10 h to obtain a silk sericin hemoglobin sponge.
[0075] Experimental determination showed that the product had antibacterial activity, high compression strength, aggregated red blood cells and rapid blood clotting properties.
[0076] Comparative Example 1
[0077] A photothermal antibacterial and hemostatic wound dressing with easy peeling and a preparation method thereof, comprising the following steps:
[0078] 500 mg of sericin protein was dissolved in 50 mL of deionized water and heated to complete dissolution, and then cooled to room temperature. Then 500 mg of carbodiimide was added and stirred for 15 min, and finally 300 mg of N-hydroxysuccinimide was added and stirred at room temperature for 2 h. The crosslinking solution was centrifuged (6000 rpm, 15 min), and the sericin sponge precursor was obtained after removing the supernatant. The precursor was pre-cooled at -80℃ for 12 h and freeze-dried for more than 10 h to obtain a sericin sponge with residual crosslinking agent. The sponge was dialyzed in deionized water for 2-3 days using a 3500 MW dialysis bag, and then pre-cooled at -80℃ for 12 h again, and freeze-dried for more than 10 h to obtain a sericin sponge.
[0079] The obtained product has weak antibacterial activity, high compression strength, aggregated red blood cells and weak blood clotting performance.
[0080] In order to illustrate the related properties of the wound dressing provided by the present application with photothermal antibacterial and easy peeling, the related properties of the wound dressing provided by the present application with photothermal antibacterial and easy peeling are described in conjunction with the drawings.
[0081] Figure 1 Fig. 1 is a scanning electron microscope image of the sericin hemoglobin dressing obtained in Examples 1-3 and an element scanning result of Example 2. Taking Example 2 as an example, the element scanning result shows that the sericin hemoglobin dressing contains iron elements, proving the existence of hemoglobin. The scanning result shows that the dressing with added hemoglobin and the comparative example both have interconnected porous structures, which is beneficial to the absorption of blood and the formation of a blood clotting barrier at the bleeding site, accelerating the blood clotting process.
[0082] Figure 2 Fig. 2 is a cyclic compression stress-strain curve of the sericin hemoglobin dressing obtained in Examples 1-3 after wetting. The sericin dressing shows a compression stress of 12.39 kPa after the first compression, and a compression stress of 10.73 kPa after 10 cycles of compression. Example 1 shows a compression stress of 16.08 kPa after the first compression, and a compression stress of 14.95 kPa after 10 cycles of compression. Example 2 shows a compression stress of 18.01 kPa after the first compression, and a compression stress of 16.35 kPa after 10 cycles of compression. Example 3 shows a compression stress of 18.37 kPa after the first compression, and a compression stress of 16.47 kPa after 10 cycles of compression. Compared with the comparative example, the hemoglobin dressing has a higher compression stress. After 10 cycles of compression, Examples 1-3 can still maintain 93.03%, 90.78, and 89.65% of the compression stress, respectively. While the comparative example can only maintain 86.60% of the compression stress after 10 cycles of compression. It is proved that the hemoglobin sericin dressing has excellent mechanical properties and can play an effective role in the blood clotting barrier during the hemostasis process.
[0083] Figure 3 Figure 7 is the antibacterial effect of the silk sericin hemoglobin dressing obtained in Examples 1-3 under 808 nm near-infrared light irradiation. As can be seen from the figure, Example 2 showed 100% sterilization effect after 3 min of near-infrared light irradiation, and Example 3 showed more excellent photothermal sterilization effect, showing 100% sterilization activity after 2 min of irradiation.
[0084] Figure 4 Figure 8 is the scanning electron microscope morphology of the silk sericin hemoglobin dressing obtained in Examples 1-3 after interaction with red blood cells. The aggregation ability of the wound dressing to red blood cells is one of the standards for evaluating the hemostatic performance of the wound dressing. As can be observed from the scanning electron microscope results, the hemoglobin silk sericin dressing has excellent aggregation effect on red blood cells, indicating its potential for high-efficiency hemostasis.
[0085] Figure 5 Figure 9 is the scanning electron microscope morphology of the silk sericin hemoglobin dressing obtained in Examples 1-3 after interaction with plasma. As can be seen from the figure, after the interaction of the hemoglobin silk sericin dressing with plasma, a dense fibrin film is formed on the surface. In contrast, in the comparative example, the silk sericin dressing interacts with plasma to form a sparse fibrin network. In comparison, the dense fibrin film has a stronger effect on aggregating red blood cells and platelets. At the same time, the smooth dense film on the surface is also conducive to the peeling of the dressing from the wound tissue.
[0086] Figure 6 Figure 10 is the statistical results of the coagulation index of the silk sericin hemoglobin dressing obtained in Examples 1-3. The coagulation index is one of the important indicators for evaluating the hemostatic ability of the wound dressing. The results show that the coagulation index of the hemoglobin silk sericin dressing and the silk sericin dressing is significantly lower than that of the blank group and the commercial gelatin group. Among them, the coagulation index of Examples 2 and 3 is significantly lower than that of other groups. It is proved that hemoglobin has a pro-coagulation effect. The dressing with high hemoglobin content has better in-vitro coagulation effect.
[0087] The preferred embodiments and their effects are described in the present application. However, once the basic creative concept is known to those skilled in the art, additional changes and modifications can be made to these embodiments. Therefore, the appended claims are intended to be interpreted as including the preferred embodiments and all changes and modifications falling within the scope of the present application.
[0088] Although the embodiments of the present application have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the present application, and the scope of the present application is defined by the appended claims and their equivalents.
Claims
1. A method of preparing a photothermally antibacterial procoagulant and easily peelable wound dressing, characterized by, The method comprises the following steps: dissolving sericin in water, then adding hemoglobin and carbodiimide in sequence and stirring uniformly, adding N-hydroxysuccinimide and stirring, and performing cross-linking reaction to obtain a cross-linking solution; applying external force to the cross-linking solution to make the activated proteins in the cross-linking solution aggregate and perform cross-linking reaction, and collecting the aggregates to obtain a hemoglobin sponge precursor; precooling the hemoglobin sponge precursor, and completely freeze-drying to obtain a sericin hemoglobin sponge; dialyzing the sericin hemoglobin sponge in water for 2-3 days, precooling again and freeze-drying to obtain a hemoglobin sericin dressing; when the external force is applied to the cross-linking solution, centrifugation is adopted to make the proteins in the cross-linking solution aggregate and perform cross-linking reaction; wherein the centrifugal speed is 1000-10000 rpm, and the centrifugal time is 1-20 min.
2. The method of preparation of photothermally antibacterial, procoagulant and easily peelable wound dressing according to claim 1, characterized in that, The mass ratio of the sericin, the carbodiimide and the N-hydroxysuccinimide is 5:1-10:1-5; The mass ratio of the sericin to the hemoglobin is 10:1-5; The cross-linking reaction temperature is 0-50℃, and the reaction time is greater than 2 h.
3. The method of preparation of photothermally antibacterial, procoagulant and easily peelable wound dressing according to claim 1, characterized in that, The external force applying mode comprises centrifugation or pressurization.
4. The method of preparation of photothermally antibacterial, procoagulant and easily peelable wound dressing according to claim 1, wherein, The precooling temperature is-80--20℃, the precooling time is 12-24 h, and the freeze-drying time is 10-24 h.
5. The method of preparation of photothermally antibacterial, procoagulant and easily peelable wound dressing according to claim 1, wherein, The molecular weight of the sericin and the hemoglobin is ≥8000 MW.
6. The method of preparation of photothermally antibacterial, procoagulant and easily peelable wound dressing according to claim 1, wherein, During the dialysis of the sericin hemoglobin sponge in water, the dialysis bag with a molecular weight cut-off of 3500 MW is adopted.
7. The method of preparation of photothermally antibacterial, procoagulant and easily peelable wound dressing according to claim 1, wherein, When the sericin is dissolved in water, boiling water bath is adopted to heat to complete dissolution, and then cooling to room temperature. 8.A wound dressing with photothermal antibacterial hemostatic and easy peeling prepared by the method of any one of claims 1-7. 9.Use of the wound dressing with photothermal antibacterial hemostatic and easy peeling of claim 8 in the preparation of antibacterial or hemostatic materials.