Maggot ointment for promoting wound healing as well as preparation method and application thereof

Abstract

The invention discloses a maggot ointment for promoting wound healing, the maggot ointment is composed of freeze-dried maggot powder, stearic acid, glycerin monostearate, triethanolamine, lauryl sodium sulfate, octadecanol, vaseline, glycerol, liquid paraffin and purified water, and the prepared ointment has anti-inflammatory, antibacterial and healing-promoting effects. The invention further provides a preparation method of the ointment. The preparation method is simple in process, low in raw material price, free of toxic and side effects and free of adverse reactions. The traditional Chinese medicine composition can effectively promote skin wound healing, relieve skin inflammation response, improve the expression level of skin epidermal growth factors and vascular endothelial growth factors and relieve scar hyperplasia.

Description

Technical Field

[0001] The invention belongs to the field of medicine for treating skin trauma, and particularly relates to a five-grain worm ointment for promoting wound healing, and a preparation method and application thereof. Background Art

[0002] Wound healing is a complex and lengthy process, including the hemostasis phase, the inflammation phase, the proliferation phase, and the remodeling phase, which ultimately forms a mature scar and restores the injured tissue. However, excessive inflammatory response may impair wound healing, leading to delayed wound closure, the formation of chronic wounds, and hypertrophic scars. Angiogenesis is one of the keys to wound healing, and the quality of angiogenesis directly affects the speed of wound healing. It not only provides oxygen and nutrients to damaged tissues, but also supports multiple healing processes such as immune response, granulation tissue formation, and epithelialization. If angiogenesis is insufficient, wound healing may be delayed and may even lead to chronic non-healing.

[0003] Existing drugs with wound healing function include antibiotics, growth factor peptides, etc., but antibiotics have certain side effects, and with the widespread use of antibiotics, bacteria are prone to drug resistance; growth factors are mostly peptide components, which have low stability, are easily inactivated, and are relatively expensive. Summary of the invention

[0004] In order to solve the above problems, the present invention provides a five-grain worm ointment for promoting wound healing. It is another purpose of the present invention to provide a preparation method and application thereof.

[0005] In order to achieve the above-mentioned object of the invention, the present invention adopts the following technical solutions:

[0006] A five-grain worm ointment for promoting wound healing is prepared from the following raw materials: 1-5g of five-grain worm freeze-dried powder, 5-10g of stearic acid, 1-5g of glyceryl monostearate, 0.1-0.5g of triethanolamine, 0.04-0.12g of sodium lauryl sulfate, 0.5-1.5g of octadecyl alcohol, 2-5mg of glycerol, 1-4g of vaseline, 6-10mg of liquid paraffin, and the rest is purified water; the sum of the above raw materials is 30g.

[0007] As further preferred embodiment of the present invention, the raw materials of the five-grain worm ointment are: 2-4g five-grain worm freeze-dried powder, 6-8g stearic acid, 1-2g glyceryl monostearate, 0.1-0.2g triethanolamine, 0.05-0.1g sodium lauryl sulfate, 0.8-1.2g octadecyl alcohol, 3-4mg glycerol, 1-2g vaseline, 6-8mg liquid paraffin, and the rest is purified water; the sum of the above raw materials is 30g.

[0008] As further preferred embodiment of the present invention, the raw materials of the five-grain worm ointment are: 3g five-grain worm freeze-dried powder, 7.2g stearic acid, 1.2g glyceryl monostearate, 0.15g triethanolamine, 0.07g sodium lauryl sulfate, 1g octadecyl alcohol, 3.9mg glycerol, 1.8g vaseline, 7.1mg liquid paraffin, and the rest is purified water; the sum of the above raw materials is 30g.

[0009] The present invention also provides a method for preparing the five-grain worm ointment for promoting wound healing, comprising the following steps:

[0010] 1) Preparation of oil phase matrix solution: Stearic acid, glyceryl monostearate, vaseline, liquid paraffin and triethanolamine are placed in container 1, heated to 80° C. in a water bath to prepare an oil phase matrix, and kept warm for later use;

[0011] 2) Preparation of aqueous matrix solution: Purified water, glycerol and octadecyl alcohol are placed in container 2, heated to 80° C. in a water bath, and then sodium dodecyl sulfate and lyophilized powder of millet worm are added to prepare an aqueous matrix;

[0012] 3) slowly adding the aqueous phase matrix into the oil phase matrix, and stirring rapidly and fully in the same direction until emulsified, thereby obtaining the five-grain worm ointment of the present invention.

[0013] As a further preferred embodiment of the present invention, in step 2), the freeze-dried grain worm powder is prepared by the following steps:

[0014] S1. Select healthy and pollution-free grain worms as raw materials, wash and screen them, remove impurities and ensure that the worm bodies are clean;

[0015] S2. Use ultraviolet light to sterilize it to ensure the safety of the insect body while maintaining its nutritional content;

[0016] S3, quickly placing the processed grain worms in a low-temperature freezing environment for pre-freezing;

[0017] S4, placing the pre-frozen grain worms in a vacuum freeze dryer for freeze-drying;

[0018] S5, the freeze-dried grain worms are finely crushed by a pulverizer to obtain fine freeze-dried powder, and then passed through an 80-mesh sieve to obtain freeze-dried grain worm powder that meets the requirements. The present invention uses an 80-mesh sieve to obtain fine powder, so that the sample maintains good gloss and ensures that the ointment is transparent and particle-free.

[0019] As a further preferred embodiment of the present invention, in step S2, the ultraviolet sterilization time is 45 minutes and the power is 30W.

[0020] As a further preferred embodiment of the present invention, in step S3, a low temperature pre-freezing below -40°C is used for 12 hours; in step S4, the freeze-drying temperature is -70°C, and the freeze-drying time is 48 hours.

[0021] The invention further discloses application of the five-grain worm ointment for promoting wound healing in preparing a medicine for promoting wound healing.

[0022] In the present invention, stearic acid, vaseline, glycerol, sodium lauryl sulfate and liquid paraffin have the effect of lubrication and moisturizing. Glyceryl monostearate and octadecyl alcohol have excellent consistency adjustment function. After being added to the ointment, it can effectively improve the viscosity of the system, making the ointment easier to apply and disperse. It has good emulsifying properties and can form a stable emulsion layer between water and oil. When preparing emulsion type ointment, adding an appropriate amount of octadecyl alcohol can effectively improve the stability of the emulsion and prevent oil-water separation. When the weight ratio of stearic acid, glyceryl monostearate and triethanolamine is 48:8:1, the emulsifying effect and the effect of use are the best.

[0023] The reason why the present invention adopts the operation of pre-freezing and then vacuum freeze-drying is that pre-freezing at a certain temperature for a certain time can ensure that the water in the insect body exists in the form of ice crystals, reduce cell tissue damage, and protect the biologically active components in the insect body. Then, vacuum freeze-drying is performed. At this time, the ice crystals formed by pre-freezing are directly sublimated into water vapor, avoiding water liquefaction, and maintaining the nutritional components and natural form of the grain mealworm to the maximum extent; moreover, the present invention finds that pre-freezing and then vacuum freeze-drying can improve the drying efficiency and significantly reduce the drying time. The grain mealworm prepared by the present invention contains more abundant biologically active components, including antimicrobial peptides and a variety of proteolytic enzymes, etc. Therefore, when preparing the ointment of the present invention, the addition amount thereof is also significantly reduced, and it has a good effect of promoting wound healing.

[0024] Compared with the prior art, the present invention has the following beneficial effects:

[0025] (1) The freeze-dried grain worm powder prepared by the present invention is not only rich in protein, fat, vitamins and minerals, but also has multiple active ingredients with pharmacological effects. It not only contains multiple essential amino acids for the human body, which are essential for tissue repair, immune regulation and metabolic balance; but also contains antimicrobial peptides and multiple proteolytic enzymes that have significant therapeutic effects on wound infection and healing.

[0026] (2) The grain insect gel provided by the present invention has a simple preparation process, low raw material price, no toxic side effects, and no adverse reactions. It can effectively promote skin wound healing, reduce skin inflammatory reactions, increase the expression levels of skin epidermal growth factor and vascular endothelial growth factor, and reduce scar hyperplasia, and has broad market prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 It is the comparison of wound healing rate results of different treatments on mice in each group;

[0028] Figure 2 These are histopathological sections of skin wounds treated with different treatments at 4, 8, 12, 16, and 20 days after injury. The sections were stained with hematoxylin and eosin;

[0029] Figure 3 This is the result of the effects of different treatments on VEGF levels during wound healing in mice. DETAILED DESCRIPTION

[0030] In order to better understand the present invention, the present invention is further described below with reference to specific embodiments.

[0031] Example 1 Preparation of Cereal Worm Freeze-dried Powder

[0032] This embodiment provides a preparation process of lyophilized corn worm powder, comprising the following steps:

[0033] S1. Select healthy and pollution-free grain worms as raw materials, wash and screen them, remove impurities and ensure that the worm bodies are clean;

[0034] S2. Use ultraviolet light to sterilize it to ensure the safety of the insect body while maintaining its nutritional content;

[0035] S3, quickly placing the processed grain worms in a low-temperature freezing environment, usually pre-freezing at below -40°C, to ensure that the water in the worm body exists in the form of ice crystals, reduce cell tissue damage, and avoid the loss of active ingredients in the worm body;

[0036] S4, placing the pre-frozen grain worms in a vacuum freeze dryer at -70°C for 48 hours. Under low temperature and low pressure conditions, ice crystals directly sublimate into water vapor, avoiding water liquefaction and preserving the nutritional components and natural form of the grain worms to the greatest extent;

[0037] S5. The freeze-dried worms are finely pulverized by an ultrafine grinder to obtain fine freeze-dried powder. The powder is sieved through an 80-mesh sieve to obtain a target powder that meets the requirements, which is used as the raw material of the worm ointment in the subsequent examples.

[0038] Embodiment 2 Grain Worm Ointment and Preparation Method

[0039] The five-grain worm ointment of the present embodiment is prepared from the following raw materials: 3g of five-grain worm freeze-dried powder, 7.2g of stearic acid, 1.2g of glyceryl monostearate, 0.15g of triethanolamine, 0.07g of sodium lauryl sulfate, 1.0g of octadecyl alcohol, 3.9mg of glycerol, 1.8g of vaseline, 7.1mg of liquid paraffin, and the rest is purified water; the sum of the above raw materials is 30g.

[0040] The preparation method comprises the following steps:

[0041] 1) Preparation of oil phase matrix solution: Stearic acid, glyceryl monostearate, vaseline, liquid paraffin and triethanolamine are placed in container 1, heated to 80° C. in a water bath to prepare an oil phase matrix, and kept warm for later use;

[0042] 2) Preparation of aqueous matrix solution: Purified water, glycerol and octadecyl alcohol are placed in container 2, heated to 80° C. in a water bath, and then sodium dodecyl sulfate and lyophilized powder of millet worm are added to prepare an aqueous matrix;

[0043] 3) slowly adding the aqueous phase matrix into the oil phase matrix, and stirring rapidly and fully in the same direction until emulsified, thereby obtaining the five-grain worm ointment of the present invention.

[0044] Example 3

[0045] The five-grain worm ointment and its preparation method in this embodiment are the same as those in Example 2, except that they are prepared from the following raw materials: 2 g of five-grain worm freeze-dried powder, 6 g of stearic acid, 1 g of glyceryl monostearate, 0.1 g of triethanolamine, 0.05 g of sodium lauryl sulfate, 0.8 g of octadecyl alcohol, 3 mg of glycerol, 1 g of vaseline, 6 mg of liquid paraffin, and the rest is purified water; the sum of the above raw materials is 30 g.

[0046] Example 4

[0047] The five-grain worm ointment and its preparation method in this embodiment are the same as those in Example 2, except that they are prepared from the following raw materials: 4 g of five-grain worm freeze-dried powder, 8 g of stearic acid, 2 g of glyceryl monostearate, 0.2 g of triethanolamine, 0.1 g of sodium lauryl sulfate, 1.2 g of octadecyl alcohol, 4 mg of glycerol, 2 g of vaseline, 8 mg of liquid paraffin, and the rest is purified water; the sum of the above raw materials is 30 g.

[0048] Example 5

[0049] The five-grain worm ointment and its preparation method in this embodiment are the same as those in Example 2, except that the raw materials of the five-grain worm ointment are: 5g of five-grain worm freeze-dried powder, 10g of stearic acid, 5g of glyceryl monostearate, 0.5g of triethanolamine, 0.12g of sodium lauryl sulfate, 1.5g of octadecyl alcohol, 5mg of glycerol, 4g of vaseline, 10mg of liquid paraffin, and the rest is purified water; the sum of the above raw materials is 30g.

[0050] Example 6

[0051] The five-grain worm ointment and its preparation method in this embodiment are the same as those in Example 2, except that the raw materials of the five-grain worm ointment are: 1g of five-grain worm freeze-dried powder, 5g of stearic acid, 1g of glyceryl monostearate, 0.1g of triethanolamine, 0.04g of sodium lauryl sulfate, 0.5g of octadecyl alcohol, 2mg of glycerol, 1g of vaseline, 6mg of liquid paraffin, and the rest is purified water; the sum of the above raw materials is 30g.

[0052] Experimental Example 1 Effect of different formula ratios of ointment base on the present invention

[0053] 1. Orthogonal experiment

[0054] This experimental example uses a (stearic acid), b (glyceryl monostearate), and c (triethanolamine) as three factors, and designs a horizontal orthogonal test, see Table 1. Other excipients in the formula are fixed as follows: 0.07g sodium lauryl sulfate, 1.0g octadecyl alcohol, 3.9mg glycerol, 1.8g vaseline, 7.1mg liquid paraffin, and purified water is added to a total of 30g. The appearance, stability, and ductility are evaluated and scored, divided into 3 levels: excellent (5 points), medium (3 points), and poor (1 point). The sum of the test results is used as an indicator to evaluate the quality of the ointment and optimize the matrix formula of the ointment.

[0055] Table 1 Orthogonal experiment table of ointment matrix formula

[0056]

[0057] 2. Evaluation of ointment matrix properties

[0058] Table 2 Orthogonal test design table of ointment matrix formula

[0059]

[0060] The influence of the three matrix factors on the quality of the ointment is a=b>c. The results are shown in Table 2. The best solution for the ointment matrix is ​​a2b3c1, that is, the best formula of the ointment is 7.2g stearic acid, 1.2g monostearate glyceryl, 0.15g triethanolamine, 0.07g sodium lauryl sulfate, 1.0g octadecyl alcohol, 3.9mg glycerol, 1.8g vaseline, 7.1mg liquid paraffin, and purified water is added to a total of 30g. The ointment of this formula has a uniform texture, no stratification after centrifugation, and is easy to apply.

[0061] Experimental Example 2: The amount of five grain insect freeze-dried powder added to the ointment prepared in Example 2 was investigated

[0062] 1. Preparation of freeze-dried powder ointment with different contents

[0063] 1.0 g (sample 1), 2.0 g (sample 2), 3.0 g (sample 3), 4.0 g (sample 4), and 5.0 g (sample 5) of freeze-dried powder were accurately weighed, and 5 portions of five-grain worm ointment with different contents were prepared according to the ointment base formula and preparation method in Example 2. The amount of extract added was determined based on the evaluation results of glossiness, appearance uniformity, centrifugal stability, and hardness.

[0064] 2. Evaluation of the properties of Wuguchong Ointment

[0065] After the freeze-dried powder is mixed with the ointment matrix, it is centrifuged at 5000r / min for 30min, and then the gloss, appearance, stability, and hardness are evaluated. When the freeze-dried powder is ≤2.0g, the sample has good appearance uniformity and centrifugal stability, but the texture is soft and there are a small number of bubbles. When the freeze-dried powder is 3.0g, the sample maintains good gloss, is transparent and particle-free, has good centrifugal stability, has uniform texture, no stratification, good coating, and no bubble generation. When the amount of freeze-dried powder added is ≥4.0g, the sample has poor gloss, poor coating ductility, and reduced centrifugal stability. Finally, the amount of freeze-dried powder added was explored, and the amount of freeze-dried powder added was determined to be 3g through evaluation of gloss, appearance, stability, and hardness.

[0066] Experimental Example 3 This experimental example uses the five grain worm ointment prepared in Example 2 as an example for efficacy evaluation 1. Establishment of mouse trauma model

[0067] 75 male Kunming mice were raised in a mouse house with a relative humidity of 40% to 60% and a temperature of (20 to 23) °C. After 7 days of adaptive feeding, the mice were randomly divided into a blank control group, a positive control group, and an experimental group. Modeling method: The mice were anesthetized with sodium pentobarbital (60 mg / kg), and the back hair was removed and adapted for one day. The mice were anesthetized with sodium pentobarbital, and after local disinfection with iodine, a circular skin with a diameter of 1 cm was aseptically removed from the side of the spine of each mouse using surgical tools. The standard was that its depth reached the deep fascia of the muscle layer to make a skin wound model. After the mouse skin wound model was successfully established, it was raised in a single cage, and sufficient water and food intake were guaranteed every day.

[0068] This experiment was divided into 3 groups, namely, blank control group (no treatment), positive control group (recombinant collagen gel, recombinant collagen gel was evenly applied to the wound every day), experimental group (grain worm ointment treatment, grain worm ointment was evenly applied to the wound every day), 25 mice in each group. The application time was 08:00-09:00, once a day. The mice were kept for 20 days, during which the weight of the mice was recorded every 4 days (the mice were fasted for 12 hours), and the wounds were photographed. The mice were fixed in a dark box for photography, and the wound diameter was measured with a ruler. The changes in the wounds were recorded on the 0th, 4th, 8th, 12th, 16th, and 20th days. The mice were free to eat and drink water during the entire experiment.

[0069] 2. Mouse Blood and Tissue Collection

[0070] On the 4th, 8th, 12th, 16th, and 20th days, 5 mice were randomly selected from each group. The mice were weighed first, and then the wounds were photographed. After that, the mice were anesthetized with sodium pentobarbital (60 mg / kg), and blood was collected from the eyeballs and skin samples were collected from the back wounds for vascular endothelial growth factor (VEGF) measurement and paraffin tissue section observation. The blood coagulated naturally for 20 minutes, centrifuged at 4°C and 3000r / min for 15 minutes in a centrifuge, and the supernatant was collected and divided into 200μL centrifuge tubes and stored in a -80°C refrigerator. The collected skin tissue was placed in 4% paraformaldehyde fixative for paraffin tissue section preparation and pathological observation. The tissue sections were observed in detail at different magnifications, and the basic pathological changes in the sections, such as congestion, congestion, hemorrhage, edema, degeneration, necrosis, hyperplasia, fibrosis, organization, granulation tissue, inflammatory changes, etc., were described and scored. The scoring criteria are shown in Table 3.

[0071] Table 3 Histopathological evaluation criteria

[0072]

[0073] Pathological tissue sections of mice in each group on days 4, 8, 12, 16, and 20 were taken for HE staining.

[0074] 3. Experimental results

[0075] like Figure 1 Within 20 days of administration, the healing rate of the experimental group was higher than that of the blank control group and the positive drug group, and there was a significant difference, indicating that it has a significant promoting effect on wound healing. This is because the grain worm has a certain antibacterial effect, and can promote the proliferation of epithelial cells and vascular endothelial cells, promote epithelialization and accelerate angiogenesis.

[0076] like Figure 2Compared with the experimental group, the blank control group had a large number of inflammatory cells infiltrating in the skin wound of mice, few new blood vessels, and incomplete re-epithelialization. As the administration time increased, the inflammatory cells in the skin of mice in the experimental group decreased significantly, the granulation tissue structure was denser, and more capillaries appeared.

[0077] Vascular endothelial growth factor (VEGF) is a highly specific vascular endothelial cell growth factor that promotes vascular permeability, extracellular matrix degeneration, endothelial cell migration, proliferation and angiogenesis.

[0078] Within 20 days of administration, if Figure 3 , VEGF content showed an upward trend, and the VEGF content in the experimental group was significantly higher than that in the blank control group, indicating that more new blood vessels and tissues appeared in the experimental group, and the blood flow of the new blood vessels and tissues helped to supply oxygen and nutrients to support the repair and cell growth that promote wound healing.

[0079] The above is only a preferred embodiment of the present invention, and does not limit the present invention in other forms. Any person skilled in the art can use the above disclosed technical content to change or modify it into an equivalent embodiment with equivalent changes. However, any simple modification, equivalent change and modification made to the above embodiment according to the technical essence of the present invention without departing from the technical solution of the present invention still belongs to the protection scope of the technical solution of the present invention.

Claims

1. A five-grain worm ointment for promoting wound healing, characterized in that: The raw materials of the five-grain worm ointment are: 1-5 g of five-grain worm freeze-dried powder, 5-10 g of stearic acid, 1-5 g of glyceryl monostearate, 0.1-0.5 g of triethanolamine, 0.04-0.12 g of sodium lauryl sulfate, 0.5-1.5 g of octadecyl alcohol, 2-5 mg of glycerol, 1-4 g of vaseline, 6-10 mg of liquid paraffin, and the rest is purified water; the sum of the above raw materials is 30 g.

2. The five-grain worm ointment for promoting wound healing according to claim 1, characterized in that: The five-grain worm ointment is prepared from the following raw materials: 2-4 g of five-grain worm freeze-dried powder, 6-8 g of stearic acid, 1-2 g of glyceryl monostearate, 0.1-0.2 g of triethanolamine, 0.05-0.1 g of sodium lauryl sulfate, 0.8-1.2 g of octadecyl alcohol, 3-4 mg of glycerol, 1-2 g of vaseline, 6-8 mg of liquid paraffin, and the rest is purified water; the sum of the above raw materials is 30 g.

3. The five-grain worm ointment for promoting wound healing according to claim 2, characterized in that: The raw materials of the five-grain worm ointment are: 3 g of five-grain worm freeze-dried powder, 7.2 g of stearic acid, 1.2 g of glyceryl monostearate, 0.15 g of triethanolamine, 0.07 g of sodium lauryl sulfate, 1 g of octadecyl alcohol, 3.9 mg of glycerol, 1.8 g of vaseline, 7.1 mg of liquid paraffin, and the rest is purified water; the sum of the above raw materials is 30 g.

4. The method for preparing the five-grain worm ointment for promoting wound healing according to any one of claims 1 to 3, characterized in that: The following steps are involved: 1) Preparation of oil phase matrix solution: Stearic acid, glyceryl monostearate, vaseline, liquid paraffin and triethanolamine are placed in container 1, heated to 80°C in a water bath to prepare an oil phase matrix, and kept warm for later use; 2) Preparation of aqueous matrix solution: Purified water, glycerol and octadecyl alcohol are placed in container 2, heated to 80°C in a water bath, and then sodium dodecyl sulfate and lyophilized powder of millet worm are added to prepare an aqueous matrix; 3) slowly adding the aqueous phase matrix into the oil phase matrix, and stirring rapidly and fully in the same direction until emulsified, thereby obtaining the five-grain worm ointment of the present invention.

5. The method for preparing the five-grain worm ointment for promoting wound healing according to claim 4, characterized in that: In step 2), the freeze-dried powder of five cereal insects is prepared by the following steps: S1. Select healthy and pollution-free grain worms as raw materials, wash and screen them, remove impurities and ensure that the worm bodies are clean; S2, sterilizing the insects with ultraviolet light to ensure the safety of the insects while maintaining their nutritional components; S3, pre-freezing the treated grain insects at low temperature; S4, placing the pre-frozen grain worms in a vacuum freeze dryer for freeze-drying; S5. The freeze-dried grain worms are finely ground by a grinder to obtain fine freeze-dried powder, which is then sieved through an 80-mesh sieve to obtain freeze-dried grain worm powder that meets the requirements.

6. The method for preparing the five-grain worm ointment for promoting wound healing according to claim 5, characterized in that: In step S2, the ultraviolet sterilization time is 45 minutes and the power is 30W.

7. The method for preparing the five-grain worm ointment for promoting wound healing according to claim 5, characterized in that: In step S3, pre-freeze at a low temperature below -40°C for 12 h, and in step S4, the freeze-drying temperature is -70°C and the freeze-drying time is 48 h.

8. Use of the five-grain worm ointment according to claim 1 in preparing a medicine for promoting wound healing.

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