Gel dressing for promoting wound healing based on scandent hop and preparation method of gel dressing

Abstract

The invention provides a gel dressing for promoting wound healing. The gel dressing is prepared by mixing a humulus scandens extracting solution, a rhizoma dioscoreae nipponicae extracting solution and a medical liquid substrate. The specific preparation process comprises the following steps: S1, taking fresh humulus japonicus and rhizoma dioscoreae nipponicae, cleaning, drying and crushing, putting humulus japonicus and rhizoma dioscoreae nipponicae powder and alcohol into an extractor, starting microwave-assisted extraction, separating and sieving after the extraction is completed, and concentrating an extracting solution to obtain a humulus japonicus-rhizoma dioscoreae nipponicae extracting solution; s2, medical carbomer powder is taken, ultrapure water is added for dissolution, the pH is adjusted to be neutral with triethanolamine, and a medical liquid substrate is obtained; s3, stirring and mixing the humulus japonicus-rhizoma dioscoreae nipponicae extracting solution and the medical liquid substrate, and standing overnight under the condition of 0-4 DEG C, so as to obtain the gel dressing. The gel dressing prepared by the invention has a remarkable curative effect on healing of diabetic wounds, can quickly heal the diabetic wounds, reduces proinflammatory factors and increases anti-inflammatory factors.

Description

Technical Field

[0001] The present invention belongs to the technical field of biomedicine, and particularly relates to a wound healing promoting gel dressing based on Humulus scandens and a preparation method thereof. Background Art

[0002] Hops and Humulus scandens flowers belong to the genus Humulus of the Cannabaceae family. The immature flower spikes with fruits of hops can be used as medicine. It is bitter in taste, slightly cool, and non-toxic; it has the effects of promoting digestion, diuresis, and calming the nerves, and is mainly used to treat indigestion, abdominal distension, edema, cystitis, pulmonary tuberculosis, cough, insomnia, leprosy and other diseases. In addition, hops can not only be used to brew beer, but also be a good fermentation agent, with important economic uses. Humulus scandens also belongs to traditional Chinese medicine, is widely distributed in China, and has rich medicinal value and good forage value. The whole plant can be used as medicine, sweet and bitter in taste, cold in nature; it has the effects of clearing heat and detoxifying, and promoting diuresis and relieving stranguria, and can treat lung heat cough, lung abscess, deficiency heat and restlessness, heat strangury, edema, dysuria, damp-heat diarrhea, toxic sores, skin pruritus and other diseases. Humulus scandens can be used as green forage, silage or sun-dried hay, and can also be used as a feed additive to improve the animal's adaptability to the environment, regulate the nutritional balance, and enhance the digestive and absorption functions.

[0003] Chinese Patent CN101904902B discloses an immune enhancer mainly composed of Humulus scandens. The Humulus scandens extract is obtained by extraction using the water extraction and alcohol precipitation method. The filtrate is filtered, concentrated, and vacuum microwave dried to obtain the Humulus scandens extract powder; it is combined with daidzein and other carriers and surfactants to form an enhancer, which can prevent and treat viral and bacterial infectious diseases. The preparation process involves a high-temperature drying process, which may reduce the content of active ingredients in the Humulus scandens extract, and the enhancer also contains daidzein, and the overall process is complex.

[0004] Therefore, for Humulus scandens with multiple effects, it is particularly necessary to further exert its medicinal effects, reduce the preparation process, and improve the medicinal value; and there is no relevant report on the treatment of wound healing using Humulus scandens at present. Summary of the Invention

[0005] In view of this, the present invention provides a wound healing promoting gel dressing based on Humulus scandens and a preparation method thereof.

[0006] To achieve the above object, the present invention adopts the following technical solutions: A wound healing promoting gel dressing, which is prepared by mixing a Humulus scandens - Dioscorea nipponica Makino extract and a medical liquid base; Among them, the Humulus scandens - Dioscorea nipponica Makino extract is obtained by extracting a mixture of Humulus scandens, Dioscorea nipponica Makino and alcohol; The medical liquid base is prepared by mixing carbomer powder and ultrapure water, and adjusting the pH to neutral with triethanolamine.

[0007] A preparation method of the above gel dressing, comprising the following steps: S1. Take fresh Humulus scandens and Dioscorea nipponica, wash, dry, crush them, place the Humulus scandens-Dioscorea nipponica powder and alcohol in an extractor, start microwave-assisted extraction, after extraction is completed, separate and sieve, concentrate the extract to obtain the Humulus scandens-Dioscorea nipponica extract; S2. Take medical carbomer powder, add ultrapure water to dissolve it, and adjust the pH to neutral with triethanolamine to obtain a medical liquid base; S3. Stir and mix the Humulus scandens-Dioscorea nipponica extract and the medical liquid base, and let it stand overnight at 0-4°C to obtain the gel dressing.

[0008] Further, the mass ratio of Humulus scandens to Dioscorea nipponica in step S1 is (7-11):(1-3).

[0009] Further, the mass ratio of the Humulus scandens-Dioscorea nipponica powder to alcohol in step S1 is 1:(20-30), and the alcohol concentration is 60%-95%.

[0010] Further, the drying temperature in step S1 is 80-95°C, and the drying time is 2-3 h.

[0011] Further, the sieve mesh number in step S1 is 30-80 meshes.

[0012] Further, the microwave power in step S1 is 480 W-550 W, and the extraction time is 15-25 min.

[0013] Further, the concentration conditions: temperature 40-75°C, pressure 10-100 mbar, rotation speed 60-120 rpm; the volume after concentration is 1 / 3-1 / 2 of the original volume.

[0014] Further, the mass-volume ratio of carbomer powder to ultrapure water in step S2 is (1-3):(100-500).

[0015] Further, the volume ratio of the Humulus scandens-Dioscorea nipponica extract to the medical liquid base in step S3 is (1-3):(1000-5000).

[0016] Compared with the prior art, the beneficial effects of the present invention are: The present invention provides a gel dressing for promoting wound healing. Through the mixed extraction of Humulus scandens and Dioscorea nipponica and the adjustment of the extraction process, the prepared gel dressing finally has excellent performance. For wound healing: the change in wound area can be reduced from 1990 μm 2 without any treatment to 605.13 μm 2, the effect is remarkable; for inflammatory factors: pro-inflammatory factors (IL-1β and TNF-α) decreased from 12.65 ng / L and 119.34 ng / L without any treatment to 10.11 ng / L and 96.2 ng / L respectively, and anti-inflammatory factors (TGF-β1 and IL-10) increased from 19.16 and 173.14 to 22.20 pg / mL and 223.58 pg / mL respectively. Detailed implementation manners

[0017] The present invention will be further described in detail below in conjunction with specific embodiments, so that those skilled in the art can understand the present invention more clearly.

[0018] The wound-healing promoting gel dressing based on Humulus scandens provided by the present invention is prepared by the following method: S1. Take fresh Humulus scandens and Dioscorea nipponica in a mass ratio of (7-11):(1-3), wash them, dry them at 80-95 °C for 2-3 h, and crush them to obtain a Humulus scandens-Dioscorea nipponica mixed powder; add the Humulus scandens-Dioscorea nipponica mixed powder to an extractor, and add alcohol with a concentration of 60%-95% and a mass 20-30 times that of the Humulus scandens-Dioscorea nipponica mixed powder, adjust the microwave to 480 W-550 W for extraction for 15-25 min; after extraction, let it stand at room temperature, filter through a sieve (30-80 mesh), and concentrate the filtrate to 1 / 3-1 / 2 of the original volume under the conditions of a temperature of 40-75 °C, a pressure of 10-100 mbar, and a rotation speed of 60-120 rpm to obtain a Humulus scandens-Dioscorea nipponica extract.

[0019] S2. Take medical carbomer powder and ultrapure water and stir and mix them according to a mass-volume ratio of (1-3):(100-500), and then add triethanolamine dropwise to adjust the pH to neutral to obtain a medical liquid base.

[0020] S3. Take the Humulus scandens-Dioscorea nipponica extract and the medical liquid base and stir and mix them according to a volume ratio of (1-3):(1000-5000), and let it stand overnight at 0-4 °C to obtain the gel dressing.

[0021] Considering from the product form, the present invention is specially prepared in the form of a gel dressing. The whole preparation process is simple and can be completed only through steps such as extraction, filtration, concentration, and mixing; in the extraction process of Humulus scandens and Dioscorea nipponica, the form of microwave can not only improve the extraction rate, but also retain the activity of the effective components to the greatest extent; finally, a wound-healing promoting gel dressing based on Humulus scandens is formed, which has a good curative effect on wound healing.

[0022] Example 1 This example provides a wound-healing promoting gel dressing based on Humulus scandens, and the specific preparation method is as follows: S1. Take 7 parts of fresh Humulus scandens and 2 parts of fresh Dioscorea nipponica, wash them, dry them at 80 °C for 3 h, and crush them with a blender to obtain Humulus scandens - Dioscorea nipponica powder; add the Humulus scandens - Dioscorea nipponica powder to an extractor, and add alcohol with a concentration of 95% and a mass 20 times that of the Humulus scandens - Dioscorea nipponica powder, adjust the microwave power to 550 W and extract for 15 min; after extraction, let it stand at room temperature for extraction and sieving (30 mesh), and concentrate the filtrate to 1 / 3 of the original volume under the conditions of a temperature of 40 °C, a pressure of 100 mbar, and a rotation speed of 60 rpm to obtain the Humulus scandens - Dioscorea nipponica extract.

[0023] S2. Take medical carbomer powder and ultrapure water, stir and mix them according to a mass - volume ratio of 1:500, and then add triethanolamine dropwise to adjust the pH to neutral to obtain a medical liquid base.

[0024] S3. Take the Humulus scandens - Dioscorea nipponica extract and the medical liquid base, stir and mix them according to a volume ratio of 3:1000, and let it stand overnight at 4 °C to obtain the gel dressing.

[0025] Example 2 This example provides a wound - promoting healing gel dressing based on Humulus scandens, and the specific preparation method is as follows: S1. Take 11 parts of fresh Humulus scandens and 3 parts of fresh Dioscorea nipponica, wash them, dry them at 90 °C for 2 h, and crush them with a blender to obtain Humulus scandens - Dioscorea nipponica powder; add the Humulus scandens - Dioscorea nipponica powder to an extractor, and add alcohol with a concentration of 60% and a mass 30 times that of the Humulus scandens - Dioscorea nipponica powder, adjust the microwave power to 480 W and extract for 25 min; after extraction, let it stand at room temperature for extraction and sieving (80 mesh), and concentrate the filtrate to 1 / 2 of the original volume under the conditions of a temperature of 75 °C, a pressure of 10 mbar, and a rotation speed of 120 rpm to obtain the Humulus scandens - Dioscorea nipponica extract.

[0026] S2. Take medical carbomer powder and ultrapure water, stir and mix them according to a mass - volume ratio of 1:100, and then add triethanolamine dropwise to adjust the pH to neutral to obtain a medical liquid base.

[0027] S3. Take the Humulus scandens - Dioscorea nipponica extract and the medical liquid base, stir and mix them according to a volume ratio of 1:5000, and let it stand overnight at 4 °C to obtain the gel dressing.

[0028] Example 3 This example provides a wound - promoting healing gel dressing based on Humulus scandens, and the specific preparation method is as follows: S1. Take 9 parts of fresh Humulus scandens and 2 parts of fresh Dioscorea nipponica, wash them, dry them at 90 °C for 2.5 h, and crush them with a wall breaker to obtain Humulus scandens-Dioscorea nipponica powder; add the Humulus scandens-Dioscorea nipponica powder to an extractor, and add alcohol with a concentration of 75% and a volume 25 times that of the Humulus scandens-Dioscorea nipponica powder, adjust the microwave power to 540 W for extraction for 20 min; after extraction, let it stand at room temperature for extraction and sieving (50 mesh), and concentrate the filtrate to 1 / 2 of the original volume under the conditions of a temperature of 60 °C, a pressure of 50 mbar, and a rotation speed of 100 rpm to obtain the Humulus scandens-Dioscorea nipponica extract.

[0029] S2. Take medical carbomer powder and ultrapure water and stir and mix them according to a mass-to-volume ratio of 1:150, then add triethanolamine dropwise to adjust the pH to neutral to obtain a medical liquid base.

[0030] S3. Take the Humulus scandens-Dioscorea nipponica extract and the medical liquid base and stir and mix them according to a volume ratio of 1:1500, and let it stand overnight at 4 °C to obtain the gel dressing.

[0031] Comparative Example 1 This example provides a wound-healing promoting gel dressing based on Humulus scandens. The raw materials and preparation method are basically the same as those in Example 3, except that: "adjust the microwave power to 540 W for extraction for 20 min" is replaced with "react in a constant temperature water bath at 45 °C for 55 minutes", and the rest remain unchanged.

[0032] Comparative Example 2 This example provides a wound-healing promoting gel dressing based on Humulus scandens. The raw materials and preparation method are basically the same as those in Example 3, except that: "adjust the microwave power to 540 W for extraction for 20 min" is replaced with "react in a constant temperature water bath at 60 °C for 55 minutes", and the rest remain unchanged.

[0033] Comparative Example 3 This comparative example provides a wound-healing promoting gel dressing based on Humulus scandens. The specific preparation method is basically the same as that in Example 3, except that: Dioscorea nipponica is not added, and Dioscorea nipponica is replaced by Humulus scandens of equal mass.

[0034] Comparative Example 4 This comparative example provides a wound-healing promoting gel dressing based on Humulus scandens. The specific preparation method is basically the same as that in Example 3, except that: Humulus scandens is not added, and Humulus scandens is replaced by Dioscorea nipponica of equal mass.

[0035] Comparative Example 5 This comparative example provides a wound-healing promoting gel dressing based on Humulus scandens. The specific preparation method is basically the same as that in Example 3, except that: Dioscorea nipponica is replaced by Nauclea officinalis of equal mass, and the rest remain unchanged.

[0036] Furthermore, in order to understand the effects of the above-mentioned examples and comparative examples, the following tests were also conducted: 1. Cell scratch assay to evaluate the wound healing-promoting ability of different extracts On the bottom surface of a six-well plate, draw three horizontal lines with a marker pen as marking lines; plate according to the grouping, with a cell density of about 5×10 5 cells / well, shake to evenly distribute the cells; after the cells grow confluent, use a pipette tip to draw three perpendicular lines perpendicular to the well plate and the drawn marking lines, so that the scratches intersect with the marking lines, forming 16 regions, and select 9 regions as fixed detection points; discard the old culture medium, gently rinse with PBS two to three times until the detached cells are rinsed clean; add the medicated culture medium (containing the extract) or serum-free culture medium according to the grouping; after completion, place in an incubator for culture, take out the cells after 24 hours, observe the scratch width at the same position under a microscope, and use ImageJ software to analyze the distance of cell migration. The specific results are shown in Table 1.

[0037] Table 1

[0038] As can be seen from the results in Table 1: Compared with the control group, Examples 3 and Comparative Examples 1-5 can all promote cell migration, but the extract obtained in Example 3 shows a better cell migration-promoting effect than the extracts obtained in other comparative examples, and has the highest potential for promoting wound healing.

[0039] 2. Wound healing-promoting experiment on diabetic foot model mice (1) Group setting a. Control group (without any treatment); b. Gel group (coated with a gel prepared from a medical liquid base without drug components); c. Gel group prepared in Example 3; d. Gel group prepared in Comparative Example 2; e. Gel group prepared in Comparative Example 3; f. Gel group prepared in Comparative Example 4; g. Gel group prepared in Comparative Example 5; (2) Experimental design Evenly distribute the diabetic foot model mice into each group according to their physical conditions, and the experimental period is 10 days. Take pictures of the wound site before and after modeling, and use ImageJ software to determine the area of the wound site. The results are shown in Table 2. After the experiment, take the tissue of the wound site for Elisa analysis of inflammatory factors. The results are shown in Table 3.

[0040] Table 2 Changes in wound area before and after different gel dressings experiments

[0041] As can be seen from Table 2, compared with the control group and the pure gel group, Examples 3 and Comparative Examples 2-5 all have a promoting effect on wound healing. Among them, the wound in Example 3 recovered best, followed by Comparative Example 3. The effects of Comparative Examples 4 and 5 were not much different, and the effect of Comparative Example 2 was the worst. Overall, the gel prepared in Example 3 has the best effect in promoting the healing of diabetic foot wounds.

[0042] Table 3 Results of wound inflammatory factors before and after experiments with different gel dressings

[0043] As can be seen from Table 3, compared with the control group and the pure gel group, Examples 3 and Comparative Examples 2-5 both have beneficial effects on the wound in reducing pro-inflammatory factors (IL-1β and TNF-α) and increasing anti-inflammatory factors (TGF-β1 and IL10). Overall, Example 3 has the best effect and is more conducive to the healing of the wound site of diabetic foot model mice.

[0044] From the above, it can be seen that this application combines two key factors, namely the ingredients and the extraction process, and finally obtains the gel dressing with the best wound healing effect.

[0045] In the present invention, the specific raw materials not described are all existing substances and can be directly purchased from the market.

[0046] The above is only a preferred implementation of the present invention and is not intended to limit the protection scope of the present invention. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principle of the present invention shall be included within the protection scope of the present invention.

Claims

1. A wound healing promoting gel dressing, characterized in that, The gel dressing is prepared by mixing the Humulus scandens-Dioscorea nipponica extract and a medical liquid base; Among them, the Humulus scandens-Dioscorea nipponica extract is prepared by extracting the mixture of Humulus scandens and Dioscorea nipponica; The medical liquid base is prepared by mixing carbomer powder and ultrapure water and adjusting the pH to neutral with triethanolamine.

2. The preparation method of the gel dressing according to claim 1, characterized in that, It includes the following steps: S1. Take fresh Humulus scandens and Dioscorea nipponica, wash, dry, crush them. Put the Humulus scandens-Dioscorea nipponica powder and alcohol into an extractor, start microwave-assisted extraction. After extraction is completed, separate and sieve, and concentrate the extract to obtain the Humulus scandens-Dioscorea nipponica extract; S2. Take medical carbomer powder, add ultrapure water to dissolve it, and adjust the pH to neutral with triethanolamine to obtain the medical liquid base; S3. Stir and mix the Humulus scandens-Dioscorea nipponica extract and the medical liquid base, and let it stand overnight at 0-4°C to obtain the gel dressing.

3. The preparation method according to claim 2, wherein In step S1, the mass ratio of Humulus scandens to Dioscorea nipponica is (7-11):(1-3).

4. The preparation method according to claim 2, characterized in that, In step S1, the mass ratio of the Humulus scandens-Dioscorea nipponica powder to alcohol is 1:(20-30), and the alcohol concentration is 60%-95%.

5. The preparation method according to claim 2, wherein In step S1, the drying temperature is 80-95°C, and the drying time is 2-3 h.

6. The preparation method according to claim 2, characterized in that, In step S1, the sieve mesh number is 30-80 meshes.

7. The preparation method according to claim 2, wherein, In step S1, the microwave power is 480W-550W, and the extraction time is 15-25 min.

8. The preparation method according to claim 2, characterized in that, Concentration conditions: temperature 40-75°C, pressure 10-100 mbar, rotation speed 60-120 rpm; the volume after concentration is 1 / 3-1 / 2 of the original volume.

9. The preparation method according to claim 2, characterized in that, In step S2, the mass-volume ratio of carbomer powder to ultrapure water is (1-3):(100-500).

10. The preparation method according to claim 2, characterized in that, In step S3, the volume ratio of the Humulus scandens-Dioscorea nipponica extract to the medical liquid base is (1-3):(1000-5000).

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