CN114159616A

CN114159616A - Liquid dressing for promoting wound repair and preparation method thereof

Info

Publication number: CN114159616A
Application number: CN202210132672.2A
Authority: CN
Inventors: 盛宏霞; 吉建飞; 高雪松; 李小刚; 陈晓颖; 李梁
Original assignee: Bosheng Zhuoyue Biological Science & Technology Beijing Co ltd
Current assignee: Bosheng Zhuoyue Biological Science & Technology Beijing Co ltd
Priority date: 2022-02-14
Filing date: 2022-02-14
Publication date: 2022-03-11

Abstract

The invention provides a liquid dressing for promoting wound repair, which comprises an exosome, a film-forming agent and a hemostatic agent. The invention also discloses a preparation method of the liquid dressing for promoting wound repair. The liquid dressing prepared by the invention has good hemostatic and antibacterial effects after the wound surface is filmed, and can effectively promote wound surface repair and accelerate wound healing.

Description

Liquid dressing for promoting wound repair and preparation method thereof

Technical Field

The invention relates to the technical field of medical dressings, in particular to a liquid dressing for promoting wound repair and a preparation method thereof.

Background

Dressings are generally referred to medically as coverings that temporarily cover the wound for protection. When the skin is damaged, a proper dressing is selected to cover the wound, so that bleeding control, infection prevention and secretion absorption are facilitated, and the wound is promoted to heal quickly. The traditional medical dressing mainly adopts cotton gauze and the like for nursing, can keep the wound surface dry and breathable and control infection. Although the cotton product is low in price and convenient to use, gaps of the cotton product are not enough to prevent most microorganisms from entering a wound healing environment, and the cotton product is easy to adhere to a wound to prevent pain and cause secondary wound. And the air permeability is not ideal, and the water vapor and sweat normally secreted by the local part of the human body can not penetrate through the air permeability, so that the soaking effect is generated on the local skin, and the phenomenon of local skin wrinkles is formed.

The chitosan is a product of partial deacetylation of natural polysaccharide chitin, is a polysaccharide substance, has good cell affinity and biocompatibility, has various physiological functions of stopping bleeding, inhibiting bacteria, promoting wound healing and the like, can be naturally degraded, and is widely applied to the fields of food, medicine, bioengineering, chemical industry and the like. The positive charges carried by the chitosan and the derivatives thereof can adsorb the cell wall with negative charges of bacteria or fungi and form a layer of polymer membrane on the cell surface, thereby changing the permeability of the cell membrane, interfering the normal metabolism of the bacteria or fungi and playing the roles of bacteriostasis and sterilization.

Exosomes are lipid membrane vesicles with the diameter of 30-100 nm secreted by cells, and play a certain role in the physiological and pathological processes of immune response, apoptosis, angiogenesis, inflammatory reaction and the like as a carrier for transmitting biological information. Researches find that the exosome, particularly the exosome from stem cells, can effectively repair the damaged tissues of the organism to achieve the aim of treating diseases. As an important component in the paracrine pathway of stem cells, the compound has better application prospect in promoting the repair and regeneration of skin wounds.

A Coloskin liquid band-aid sold on the market. The product has short film forming time and good water resistance. In the formula, volatile solvent isopropanol is used, and a large amount of cellulose nitrate is used as a film forming material. But the product has heavy pain and poor use feeling in the use process.

Chinese patent CN110975004A discloses a bacteriostatic liquid band-aid capable of quickly forming a film. The liquid band-aid prepared by the invention has short film forming time and good resilience, is added with antibacterial components and has antibacterial effect. However, the liquid band-aid of the invention does not contain components capable of promoting wound healing and has no wound healing promoting effect.

Disclosure of Invention

In order to solve the technical problems, the invention provides a liquid dressing for promoting wound repair and a preparation method thereof. The liquid dressing has good film forming property and antibacterial property, can be directly applied to the wound surface of skin, and can effectively prevent wound surface infection, prevent wound surface persistent hemorrhage, and promote wound surface healing.

In a first aspect of the invention, a liquid dressing for promoting wound repair is provided, the liquid dressing comprising exosome, a film-forming agent and a hemostatic agent.

Preferably, the content of exosomes in the liquid dressing is 10-50%, and the exosomes are derived from human umbilical cord mesenchymal stem cells. More preferably, the exosome has a particle size of 50-300 μm and a particle number of 2 × 10¹⁰/ml。

Preferably, the content of the film forming agent in the liquid dressing is 20-65%, and the film forming agent comprises one or more of water-soluble chitosan, poloxamer, polyvinyl alcohol, polyvinylpyrrolidone, collagen and gelatin. More preferably, the film forming agent comprises water-soluble chitosan and poloxamer, the weight ratio of the water-soluble chitosan to the poloxamer is 1 (1-15), further, the weight ratio of the water-soluble chitosan to the poloxamer is 1 (8-15), 1 (10-15), 1 (12-15); further preferably, the poloxamers include poloxamer 407 and poloxamer 188, the ratio of poloxamer 407: poloxamer 188 is present in a weight ratio of (1-3) to (3-1), such as 3:1, 2:1, 1:1, 1:2, 1:3, and so forth.

Preferably, the content of the hemostatic agent in the liquid dressing is 0.1-1.5%, and the hemostatic agent comprises glycine.

Preferably, the liquid dressing further comprises water, the water content being 5-30%.

In a specific embodiment, the liquid dressing comprises 1-50% exosomes, 1-15% water-soluble chitosan, 5-30% poloxamer 407, 5-30% poloxamer 188, 0.1-0.5% glycine and 5-30% water.

More preferably, the liquid dressing comprises 3% water-soluble chitosan, 36% exosomes, 18% poloxamer 407, 18% poloxamer 188, 0.5% glycine and 24.5% water.

More preferably, the liquid dressing comprises 15% water-soluble chitosan, 42% exosomes, 7.5% poloxamer 407, 7.5% poloxamer 188, 0.1% glycine and 27.9% water.

The percentages are by weight except for the exosomes by volume.

In a second aspect of the present invention, there is provided a method for preparing the liquid dressing, the method comprising:

1) dissolving the film forming agent and the hemostatic agent in proportion, and mixing to obtain uniform and transparent mixed liquid A;

2) and adding exosome in a certain proportion into the mixed solution A, and uniformly stirring to obtain the liquid dressing for promoting wound repair.

Preferably, the preparation method comprises the following steps:

1) dissolving water-soluble chitosan in purified water according to a ratio, standing for defoaming, adding glycine, stirring for dissolving, adding poloxamer 188 and poloxamer 407, and incubating overnight at 4 ℃ to obtain uniform and transparent mixed liquid A;

2) and adding the exosome into the mixed solution A, uniformly stirring, and preparing to obtain the liquid dressing for promoting wound repair.

The invention has the beneficial effects that:

(1) the film forming agent in the liquid dressing contains chitosan, and is combined with poloxamer 188 and poloxamer 407, so that the liquid dressing has good film forming property and good ductility. Compared with other liquid dressings, the composition is not influenced by the shape and size of a wound when used, and has the property of fast self-filming.

(2) The bacteriostatic agent in the liquid dressing comprises chitosan, and the chitosan has better biodegradability and bacteriostatic property. Compared with the common bacteriostatic agent, the liquid auxiliary material has the advantages of wide antibacterial spectrum, high sterilization rate, no toxicity, no side effect, and capability of efficiently inhibiting the growth and reproduction of bacteria and fungi, and simultaneously, the chitosan and other film forming agents have the performance of fast self-film forming, so that the liquid auxiliary material has better antibacterial effect without additionally adding other antibacterial agents, such as antibacterial peptide and the like.

(3) The liquid dressing disclosed by the invention comprises the exosome, and the exosome can promote the proliferation and migration of skin fibroblasts, influence the functions of epidermal cells, promote the re-epithelialization of a wound surface and achieve the purpose of wound surface tissue repair.

(4) The hemostatic agent in the liquid dressing comprises glycine, and can be used for local hemostasis.

(5) The adhesive bandage is transparent, uniform and free of peculiar smell, and has good appearance and experience.

(6) The composite material is weakly acidic, can be directly applied to wound surfaces of small wounds, bruises, cut wounds and the like, is free from pain and has good safety.

Drawings

Fig. 1 is a graph showing the effect of film formation by liquid dressing, and fig. 1(a), 1(b), and 1(c) correspond to a liquid dressing sample 1 group, a liquid dressing sample 2 group, and a liquid dressing sample 3 group, respectively.

Fig. 2 is a graph showing the results of the bacteriostatic experiments performed on the liquid dressings, and fig. 2(a), 2(b), and 2(c) correspond to the liquid dressing sample 1 group, the liquid dressing sample 2 group, and the blank control group, respectively.

Fig. 3 shows the results of the experiment performed on day 0 of the wound healing experiment of mice with the liquid dressing of the present invention, and fig. 3(a), 3(b), and 3(c) correspond to sample 1, sample 3, and blank control, respectively.

Fig. 4 shows the results of the experiment performed on day 3 of the wound healing experiment of mice using the liquid dressing of the present invention, and fig. 4(a), 4(b), and 4(c) correspond to sample 1, sample 3, and blank control, respectively.

Fig. 5 shows the results of the experiment performed on the 7 th day of the wound healing experiment of mice with the liquid dressing of the present invention, and fig. 5(a), 5(b), and 5(c) correspond to sample 1, sample 3, and blank control, respectively.

Fig. 6 shows the results of the experiment performed on the 10 th day of the wound healing experiment of mice using the liquid dressing of the present invention, and fig. 6(a), 6(b), and 6(c) correspond to sample 1, sample 3, and blank control, respectively.

Fig. 7 shows the results of the experiment performed on day 15 in the wound healing experiment of mice using the liquid dressing of the present invention, and fig. 7(a), 7(b), and 7(c) correspond to sample 1, sample 3, and blank control, respectively.

Detailed Description

The invention will be further described with reference to specific embodiments, and the advantages and features of the invention will become apparent as the description proceeds. These examples are illustrative only and do not limit the scope of the present invention in any way. It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention, and that such changes and modifications may be made without departing from the spirit and scope of the invention.

It should be noted that the main chemical reagents and the manufacturer information thereof in the present invention are: water-soluble chitosan (beijing couscoibles technologies, ltd.), exosomes (platinogenesis excellent biotechnology (beijing) ltd.), poloxamer 407, poloxamer 188 (beijing solibao technologies, ltd.), glycine (shanghai lin biochemistry technologies, ltd.). The rest of the reagents are conventional reagents.

Example 1: preparation of liquid dressing samples

1. Preparation of liquid dressing sample 1

1.1 weighing the materials according to the mixture ratio in the table 1.

Table 1: material proportioning

1.2 preparation of liquid dressings

The specific preparation process of the liquid dressing comprises the following steps:

step 1): dissolving water-soluble chitosan in purified water, standing for defoaming, adding glycine, stirring for dissolving, adding poloxamer 188 and poloxamer 407, and incubating overnight at 4 ℃ to obtain uniform and transparent mixed liquid A;

step 2): and adding the exosome into the mixed solution A, uniformly stirring, and preparing to obtain a liquid dressing sample 1 for promoting wound repair.

2. Preparation of liquid dressing sample 2

2.1 weighing the materials according to the mixture ratio in the table 2.

Table 2: material proportioning

2.2 preparation of liquid dressings

step 2): and adding the exosome into the mixed solution A, uniformly stirring, and preparing to obtain a liquid dressing sample 2 for promoting wound repair.

3. Preparation of liquid dressing sample 3

3.1 weighing the materials according to the mixture ratio in the table 3.

Example 3: preparation of Chitosan reagent

Table 3: material proportioning

3.2 preparation of liquid dressings

Dissolving water-soluble chitosan in purified water, standing for defoaming, adding aminoacetic acid, stirring and dissolving to obtain a liquid dressing sample 3.

EXAMPLE 2 film Forming Properties of liquid dressing samples

In order to investigate the film forming property of the liquid dressing prepared by the method, a film forming test is carried out on liquid dressing samples 1-3, and the specific method comprises the following steps:

an appropriate amount of a liquid dressing sample was applied to any area of the skin, waiting for film formation, and the film formation effect was observed (see fig. 1) while the film formation time was recorded, and the results are shown in table 4.

Table 4: film forming time of liquid dressing with different compositions

The results in table 4 show that the film forming time of the liquid dressing sample 1 prepared by the present invention is shorter than that of the liquid dressing sample 2, while the liquid dressing sample 3 cannot form a film.

In addition, the results of fig. 1 show that both the liquid dressing samples 1 and 2 prepared according to the present invention were able to form films, whereas the liquid dressing sample 3 was not able to form films.

The results in table 4 and figure 1 demonstrate that water-soluble chitosan alone does not enable film formation of a liquid dressing. However, the water-soluble chitosan and the poloxamer are matched with each other, so that the liquid dressing can be rapidly self-filmed. The film forming effect of the liquid dressing can be adjusted by adjusting the proportion of the water-soluble chitosan and the poloxamer, and the higher the relative proportion of the poloxamer is, the better the film forming property is. Meanwhile, the antibacterial effect of the water-soluble chitosan is comprehensively considered, and preferably, the ratio of the two is as follows: the water-soluble chitosan is poloxamer in a ratio of 1 (1-15), wherein poloxamer 407 is poloxamer 188 in a ratio of (1-3) to (3: 1).

Example 3: bacteriostatic effect of liquid dressing sample

In order to investigate the bacteriostatic performance of the liquid dressing prepared by the method, a liquid dressing sample 1 and a liquid dressing sample 2 are selected to carry out a bacteriostatic test experiment, and the specific process is as follows:

step 1: and inoculating the activated escherichia coli bacterial suspension on an LB culture medium, and carrying out streaking operation.

Step 2: and (3) respectively putting the sterilized circular filter paper sheets into the liquid dressing sample 1 and the liquid dressing sample 2 groups for soaking, wherein the blank control group does not soak the liquid dressing. The soaked sterilized round filter paper sheets are respectively taken by sterile tweezers and pasted in different culture dishes for culture at 37 ℃ for 18-24 h, and the bacteriostasis result is shown in figure 2.

The results in FIG. 2(a) show that substantially no bacteria were visible to the naked eye on the media of the liquid dressing sample group 1. The results in FIG. 2(b) show that a small amount of bacteria was visible to the naked eye on the medium of the liquid dressing sample 2 group. The results in FIG. 2(c) show that a large number of bacteria were visually observed on the medium of the blank control group, which was significantly different from those of the sample 1 group and the sample 2 group. The liquid dressing prepared by the invention has obvious bacteriostatic action, and the bacteriostatic effect of the liquid dressing sample 1 is better than that of the liquid dressing sample 2, probably because the film-forming property of the liquid dressing sample 1 is better than that of the liquid dressing sample 2, the liquid dressing sample can form a film at a higher speed, and a wound is isolated from the external environment. In the preparation process, although the proportion of chitosan in the group 2 is higher, the prepared liquid dressing has relatively poor properties, and the bacteriostatic effect of the liquid dressing can be influenced to a certain extent, so that various performances of the liquid dressing are mutually supported in function and cannot be singly divided.

The results also prove that the combination of chitosan and other film-forming agents has rapid self-film-forming performance, so that the liquid auxiliary material disclosed by the invention has a good antibacterial effect without additionally adding other antibacterial agents, such as antibacterial peptide and the like.

Example 4: effect of liquid dressing samples on wound healing

A liquid dressing sample 1 and a liquid dressing sample 3 were selected for a mouse wound healing experiment and labeled as sample 1 group and sample 3 group, respectively, while the treatment without application of a liquid dressing sample was used as a blank control group. The specific experimental process is as follows:

mice weighing about 20 g were used. After general anesthesia of mice, the back of the mice is shaved, and after disinfection, epidermal skin is cut from the back of the mice by using a round medical puncher with the diameter of 1 cm, each group comprises 2 mice, and the mice are raised in cages to avoid mutual licking and rubbing of the mice. Sample 1, group 1 and sample 3 groups were applied with liquid dressing sample 1 and liquid dressing sample 3, respectively, daily, and the blank control group was not applied with liquid dressing. In the process of breeding the mice, the wound healing condition of each mouse is observed on the 0 th day, the 3 rd day, the 7 th day, the 10 th day and the 15 th day, and the wound healing process is recorded, as shown in fig. 3-7.

As can be seen from fig. 3 to 7, in the process of wound healing, the wounds of the sample 1 group and the sample 3 group are healed obviously on day 3, wherein the wound healing rates of the sample 1 group, the sample 3 group and the blank control group are reduced in sequence (see fig. 4). The wounds had scabbed on day 7, and the healing progress was consistent among the groups with no apparent difference between the groups (see figure 5). On day 10, the wounds of sample 1 and sample 3 were substantially healed, with the wound healing of sample 1 being significantly better than that of sample 3 and the wound healing of the blank control being slower (see fig. 6). The smoothness of the wound surface of the sample 1 group is better than that of the sample 3 group on the 15 th day, and the wound area is better healed. The wound surface of the sample 3 group healed, but the wound surface is obviously not smooth enough. The blank control group did not heal completely (see fig. 7).

The results of fig. 3 to 7 show that both the liquid dressing sample 1 and the liquid dressing sample 3 can promote the healing of the wound compared to the blank control group. However, the speed and effect of promoting wound healing of the liquid dressing sample 1 were superior to those of the liquid dressing sample 3. The addition of the exosome and the poloxamer can improve the capability of the liquid dressing in promoting the repair and regeneration of skin wounds and optimize the performance of the liquid dressing.

Although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that changes may be made in the embodiments and/or equivalents thereof without departing from the spirit and scope of the invention. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. The liquid dressing for promoting wound repair is characterized by comprising 10-50% of exosome, 20-65% of film-forming agent and 0.1-1.5% of hemostatic agent, wherein the film-forming agent comprises water-soluble chitosan and poloxamer.

2. The liquid dressing of claim 1, wherein the weight ratio of water-soluble chitosan to poloxamer is 1 (1-15).

3. The liquid dressing of claim 1, wherein the poloxamers comprise poloxamer 407 and poloxamer 188.

4. The liquid dressing of claim 3, wherein the weight ratio of poloxamer 407 to poloxamer 188 is (1-3) to (3-1).

5. The liquid dressing of claim 1, wherein the hemostatic agent comprises glycine.

6. The liquid dressing of claim 1, wherein the liquid dressing comprises 10-50% exosomes, 1-15% water-soluble chitosan, 5-30% poloxamer 407, 5-30% poloxamer 188, 0.1-0.5% glycine and 5-30% water.

7. A method of manufacturing a liquid dressing according to any one of claims 1 to 6, wherein the method of manufacturing comprises:

1) dissolving the film forming agent and the hemostatic agent according to a certain proportion, and mixing to obtain uniform and transparent mixed liquid A;

2) and adding the exosome in a certain proportion into the mixed solution A, and uniformly stirring to obtain the liquid dressing for promoting wound repair.