Core-shell structure particle for promoting wound healing and application thereof

Abstract

The invention belongs to the field of biomedical dressings, and discloses core-shell structure particles for promoting wound healing and application of the core-shell structure particles. The core-shell structure particles are obtained by taking a trehalose solution and a lucid ganoderma source chitosan solution as a core solution and a membrane casting solution respectively, carrying out air bath and coagulating bath in a pulse mode, and separating. The core-shell structure particles of some examples have relatively stable structures, can be stably dispersed in aqueous materials, and can be prepared into wound healing preparations in dosage forms, such as dressings, sprays, pastes and the like. The particle size of the core-shell structure particles is adjustable, so that the core-shell structure particles can go deep into the wound surface to realize better wound surface coverage and better promote wound surface healing. According to the core-shell structure particles of some examples, various repairing factors can be loaded in the shell, controlled release and slow release of the repairing factors can be achieved due to existence of the shell, and a better repairing effect is achieved.

Description

Technical Field

[0001] The invention belongs to the field of biomedical dressings, and in particular relates to core-shell structure particles for promoting wound healing and applications thereof. Background Art

[0002] With the development of medical technology, gel dressings are increasingly used, and they have many advantages over traditional gauze dressings. As a commonly used wound dressing, gel dressings can absorb the exudate from the patient's wound and keep the wound moist and clean, thereby providing a good healing environment for the wound. They are suitable for superficial and non-chronic wounds.

[0003] For non-chronic wounds on certain facial or exposed areas, such as superficial wounds, first-degree or shallow second-degree burn wounds, surgical suture wounds, and wounds formed after laser / photon / acid peeling / microneedle treatment of micro plastic surgery, patients or users have more stringent requirements for the repair time and repair effect of the dressing, and the dressing should not adhere to the wound when removed, causing secondary damage to the wound. In addition, users also have high expectations for the safety of the dressing ingredients and the user experience.

[0004] Chitosan is the deacetylated product of chitin. It is generally believed that chitosan is chitosan when the deacetylation degree of chitin is greater than 55%. The structure of chitosan is very similar to that of cellulose. Both are composed of multiple glucose molecules connected by glycosidic bonds. The difference is that chitosan contains a large number of free amino groups. The large number of active groups in chitosan give it many functions. For example, the large number of polar groups in chitosan give it hydrophilic and moisturizing properties, making it suitable for moisturizing skin care products; it can activate macrophages, T lymphocytes, and NK cells, and then activate the immune system, and has anti-tumor activity; in addition, chitosan can reduce the levels of cholesterol and triglycerides in the blood and prevent cardiovascular and cerebrovascular diseases; as an alkaline natural polysaccharide, it also has anti-gastric acid and anti-ulcer effects; at the same time, it can also play a regulatory role in the treatment of allergic dermatitis, reducing the levels of serum cholesterol, urea and creatine in patients with kidney disease, etc. Compared with traditional chitosan derived from shrimp and crab shells, chitosan derived from Ganoderma lucidum has the characteristics of smaller molecular weight and no allergenic protein. In addition, chitosan from Ganoderma lucidum is usually covalently linked to dextran. Dextran has hydrophilicity, high adsorption performance and good biocompatibility, and has been widely used in medical dressings such as skin care, drug sustained release and wound healing. In recent years, the dextran-chitin complex extracted from Ganoderma lucidum has attracted widespread interest among researchers.

[0005] Chitosan has certain antibacterial properties. Studies have shown that its antibacterial properties are affected by pH. Under acidic conditions, chitosan exhibits cationic properties. Electrostatic interactions can occur between the positively charged chitosan and the negatively charged bacterial cell membrane, changing the permeability of the bacterial cell membrane, thereby causing bacterial cell death and exerting an antibacterial effect. Chitosan is a cationic polyamine that is slightly soluble in water and almost insoluble in ethanol. This results in existing chitosan dressings being basically formed dressings, or chitosan being modified to increase its solubility. This limits the scope of application of chitosan in dressings, or may lead to increased costs.

[0006] Trehalose is a non-reducing disaccharide composed of two glucose molecules connected by an α,1,1-glycosidic bond. Trehalose is very stable to heat and acid and is one of the most stable sugars among natural disaccharides. It is not easy to decompose at high temperatures, nor is it easy to undergo Maillard reaction with amino acids, proteins, etc. Trehalose has good solubility in aqueous solution and good hygroscopicity. Its aqueous solution is not easy to brown at high temperatures. Trehalose is a common moisturizer in dressings.

[0007] There is a significant difference in the solubility of chitosan and trehalose, which makes it difficult to mix the two evenly, which is not conducive to the preparation of a dressing with a uniform texture. Summary of the invention

[0008] The object of the present invention is to overcome at least one deficiency of the prior art and provide a core-shell structure particle for promoting wound healing and application thereof.

[0009] The technical solution adopted by the present invention is: The first aspect of the present invention provides: A core-shell structure particle for promoting wound healing, wherein the preparation method comprises the following steps: Prepare trehalose solution and ganoderma lucidum chitosan solution respectively; Trehalose solution is used as the core liquid, and Ganoderma lucidum chitosan solution is used as the casting liquid. The solution is pulsed through the spinning head, and the droplets fall into the alkaline coagulation bath after passing through the air bath. The core-shell structure particles are obtained by separation and washing.

[0010] In some examples of core-shell structured particles, the length of the air bath is 10 to 20 cm.

[0011] In some examples of core-shell structured particles, the mass concentration of the trehalose solution is 30-45%.

[0012] In some examples of core-shell structured particles, the mass concentration of the Ganoderma lucidum source chitosan solution is 2-5%.

[0013] In some examples of core-shell structured particles, the mass ratio of the trehalose solution to the ganoderma lucidum source chitosan solution is 1:(3-8).

[0014] In some examples of core-shell structured particles, molecules having a repairing effect on wound surfaces are also added to the trehalose solution.

[0015] In some examples of core-shell structured particles, the pH of the trehalose solution is 5.0-6.8.

[0016] In some examples of core-shell structured particles, the coagulation bath is a NaOH solution or an ethanol solution with NaOH added.

[0017] The above features can be combined arbitrarily unless they conflict with each other.

[0018] The second aspect of the present invention provides: A dressing for promoting wound healing, which is added with the core-shell structure particles described in the first aspect of the present invention.

[0019] In some examples of dressings, the dressing is a hydrogel, a spray, a patch, an ointment, etc.

[0020] The beneficial effects of the present invention are: The core-shell structure particles of some examples of the present invention have a relatively stable structure and can be stably dispersed in aqueous materials, and can be used to prepare dosage forms of wound healing preparations, such as dressings, sprays, ointments, etc. The particle size of the core-shell structure particles is adjustable, and they can penetrate deep into the wound surface, achieve more comprehensive wound coverage, and better promote wound healing.

[0021] In some examples of the core-shell structured particles of the present invention, a variety of repair factors can be loaded in the shell. The presence of the shell can also achieve controlled and sustained release of the repair factors, thereby achieving a better repair effect. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 This is a photograph of a mouse wound healing experiment of a dressing prepared with core-shell structure particles in Example 1.

[0023] Figure 2 to Figure 5 This is a photo of the skin recovery of a scald patient using a dressing prepared with the core-shell structured particles of Example 2. DETAILED DESCRIPTION

[0024] The first aspect of the present invention provides: A core-shell structure particle for promoting wound healing, wherein the preparation method comprises the following steps: Prepare trehalose solution and ganoderma lucidum chitosan solution respectively; Trehalose solution is used as the core liquid, and Ganoderma lucidum chitosan solution is used as the casting liquid. The solution is pulsed through the spinning head, and the droplets fall into the alkaline coagulation bath after passing through the air bath. The core-shell structure particles are obtained by separation and washing.

[0025] In some examples of core-shell structured particles, the length of the air bath is 10 to 20 cm. The presence of the air bath allows the droplets to shrink into microparticles. Experimental data show that an air bath of 10 to 20 cm can obtain core-shell structured particles with more uniform particle sizes, and trehalose can be well wrapped in the shell formed by chitosan.

[0026] In some examples of core-shell structure particles, the mass concentration of the trehalose solution is 30-45%. Data show that trehalose solution of this concentration is conducive to obtaining core-shell structure particles with a more complete structure.

[0027] In some examples of core-shell structured particles, the mass concentration of the Ganoderma lucidum source chitosan solution is 2-5%.

[0028] In some examples of core-shell structured particles, the mass ratio of the trehalose solution to the ganoderma lucidum source chitosan solution is 1:(3-8).

[0029] In some examples of core-shell structured particles, the trehalose solution is further added with molecules that have a repairing effect on the wound surface. The effective molecules include but are not limited to type I collagen.

[0030] In some examples of core-shell structured particles, the pH of the trehalose solution is 5.0-6.8, which is close to the physiological pH of the skin and is more conducive to skin recovery.

[0031] In some examples of core-shell structured particles, the coagulation bath is a NaOH solution or an ethanol solution with NaOH added.

[0032] The above features can be combined arbitrarily unless they conflict with each other.

[0033] The second aspect of the present invention provides: A dressing for promoting wound healing, which is added with the core-shell structure particles described in the first aspect of the present invention.

[0034] In some examples of dressings, the dressing is a hydrogel, a spray, a patch, an ointment, etc.

[0035] The technical solution of the present invention is further explained below in conjunction with examples.

[0036] Embodiment 1: The chitosan from Ganoderma lucidum was dissolved in acetic acid solution to prepare a chitosan solution with a mass concentration of 4%, and vacuum degassing was performed; Dissolve trehalose in water to prepare a trehalose solution with a mass concentration of 40% and a pH of 6.0; The trehalose solution is used as the core liquid, the ganoderma lucidum chitosan solution is used as the casting liquid, the mass ratio of the trehalose solution to the ganoderma lucidum chitosan solution is 1:5, the core liquid is inside, the casting liquid is outside, and they pass through the spinning head together in a pulsed manner. After the droplets pass through a 15 cm long air bath, they fall into an alkaline coagulation bath, and the coagulation bath is a 60 v / v% ethanol solution added with 1 wt.% NaOH; The core-shell structure particles are obtained by filtering, separating and washing.

[0037] Embodiment 2: The chitosan from Ganoderma lucidum was dissolved in acetic acid solution to prepare a chitosan solution with a mass concentration of 5%, and vacuum degassing was performed; Dissolve trehalose in water to prepare a trehalose solution with a mass concentration of 30% and a pH of 5.0; The trehalose solution is used as the core liquid, the ganoderma lucidum chitosan solution is used as the casting liquid, the mass ratio of the trehalose solution to the ganoderma lucidum chitosan solution is 1:3, the core liquid is inside, the casting liquid is outside, and they pass through the spinning head together in a pulsed manner. After the droplets pass through a 10 cm long air bath, they fall into an alkaline coagulation bath, and the coagulation bath is a 70 v / v% ethanol solution added with 0.5 wt.% NaOH; The core-shell structure particles are obtained by filtering, separating and washing.

[0038] Embodiment 3: The chitosan from Ganoderma lucidum was dissolved in acetic acid solution to prepare a chitosan solution with a mass concentration of 3%, and vacuum degassing was performed; Dissolve trehalose in water to prepare a trehalose solution with a mass concentration of 45% and a pH of 6.8; Trehalose solution is used as core liquid, Ganoderma lucidum chitosan solution is used as casting liquid, the mass ratio of trehalose solution to Ganoderma lucidum chitosan solution is 1:6, the core liquid is inside, the casting liquid is outside, and they pass through the spinning head together in a pulsed manner. After the droplets pass through a 20 cm long air bath, they fall into an alkaline coagulation bath, and the coagulation bath is a 60 v / v% ethanol solution added with 1.0 wt.% NaOH; The core-shell structure particles are obtained by filtering, separating and washing.

[0039] Embodiment 4: Dissolve ganoderma lucidum chitosan in acetic acid solution to prepare a ganoderma lucidum chitosan solution with a mass concentration of 2%, and perform vacuum degassing; Dissolve trehalose in water to prepare a trehalose solution with a mass concentration of 35% and a pH of 5.5; The trehalose solution is used as the core liquid, the ganoderma lucidum chitosan solution is used as the casting liquid, the mass ratio of the trehalose solution to the ganoderma lucidum chitosan solution is 1:8, the core liquid is inside, the casting liquid is outside, and they pass through the spinning head together in a pulsed manner. After the droplets pass through an 18 cm long air bath, they fall into an alkaline coagulation bath, and the coagulation bath is a 50 v / v% ethanol solution added with 1.5 wt.% NaOH; The core-shell structure particles are obtained by filtering, separating and washing.

[0040] Comparative Example 1: Same as Example 1, except that no air bath treatment was performed.

[0041] Comparative Example 2: Same as Example 1, except that the mass concentration of the Ganoderma lucidum source chitosan solution is 1%.

[0042] Comparative Example 3: Same as Example 1, except that the mass concentration of the trehalose solution is 25%.

[0043] Comparative Example 4: Same as Example 1, except that the coagulation bath is 40 v / v% ethanol.

[0044] Comparative Example 5: The same as Example 1, except that the mass ratio of the trehalose solution to the ganoderma lucidum chitosan solution is 1:1.

[0045] Performance testing of core-shell structure particles: The particles prepared in each example were taken to test their appearance, particle size, and suspension stability in water. The appearance was observed using a microscope.

[0046] The test method for suspension stability is as follows: add the particles in a ratio of 1 wt% to physiological saline, mix well and let stand, test the suspension stability of the particles in physiological saline, repeat 3 times and take the average value.

[0047] The test results are shown in Table 1.

[0048] Table 1. Detection results of different example particles

[0049] Mouse wound healing experiment To further test the repair effect of the prepared core-shell structure particles on the skin, they were prepared into a dressing. The preparation method of the dressing is as follows: take 10 parts by mass of the core-shell structure particles prepared in Examples 1 to 4, Comparative Example 4 and Comparative Example 5, 3 parts by mass of glycerol, 0.25 parts by mass of carbomer, 0.15 parts by mass of triethanolamine, 0.3 parts by mass of methylparaben, add purified water to 100 parts by mass, and mix well. The effect of the dressing on wound healing was tested using a mouse wound healing experiment. The specific method is as follows: SPF mice, female, 42. After the quarantine, a 2.0 cm × 1.0 cm full-thickness skin defect wound was created on the back of anesthetized mice under sterile conditions. The depth was based on reaching the fascia, and the damaged tissue was leaking tissue fluid. The skin wound model was induced, and wound photos were taken and the wound area was recorded, which was the area d0 at this time. The model mice were randomly divided into a model control group, an embodiment group, and a control group, with 6 mice in each group. The mice in the model control group were treated with an equal amount of normal saline, and the embodiment group and the control group were treated with smear dressings. Once a day, apply continuously for 15 days. The wounds of the mice were photographed (with a ruler) and measured on the 3rd, 6th, 9th, 12th, and 15th days, and the wound area was measured using software. The wound areas were recorded as area D3, area D6, area D9, area D12, and area D15, respectively.

[0050] The test samples were applied daily, and on the 0th, 3rd, 6th, 9th, 12th and 15th days after surgery, the wound color changes, exudate, granulation tissue, growth and epithelial growth were observed to evaluate the wound healing effect and calculate the wound contraction rate (%).

[0051] Wound healing rate (%) = (area D0-area Dn) / area D0×100%; The area Dn refers to the wound area on the nth day after surgery.

[0052] All data were expressed as (x±s), and SPSS21.0 software was used for statistical analysis; one-way ANOVA was used for statistical analysis. The test results are shown in Table 2.

[0053] Table 2. Wound healing rate (%)

[0054] As shown in Table 2, Examples 1 to 4 can better promote wound healing, and the dressing of Example 2 even shortens the wound healing time to 9 days. It can be seen that the particle size of the core-shell structure particles has an unexpected effect on wound healing.

[0055] Figure 1 This is a photograph of the experimental results of Example 1.

[0056] In compliance with medical ethics and with informed consent, on the third day after the drug treatment, when there was no visible bleeding point on the wound, the dressing of Example 2 was used for auxiliary treatment of a patient with arm burns. During the auxiliary treatment, the dressing was applied to the wound surface once in the morning and evening every day. The results are as follows: Figure 2 to Figure 5 As shown. Among them, Figure 2 This is a photo taken on the third day after drug treatment for burns and before auxiliary treatment. It can be seen that the skin at the wound is obviously damaged, the surrounding skin is a little red, and the edges of the damaged skin are slightly raised, revealing the brighter red new skin tissue underneath. Figure 3 This is a photo taken 3 days after auxiliary treatment. The edges of the wound began to scab and peel off. Figure 4 This is a photo taken 5 days after auxiliary treatment. The skin is healing and gradually recovering. Figure 4 This is a photo taken 7 days after auxiliary treatment. It can be seen that the redness has been reduced and the skin is in good condition of repair.

[0057] The above is a further detailed description of the present invention, which should not be regarded as a limitation on the specific implementation of the present invention. For ordinary technicians in the technical field to which the present invention belongs, simple deduction or replacement without departing from the concept of the present invention is within the protection scope of the present invention.

Claims

1. A core-shell structure particle for promoting wound healing, wherein the preparation method comprises the following steps: Prepare trehalose solution and ganoderma lucidum chitosan solution respectively; Trehalose solution is used as the core liquid, and Ganoderma lucidum chitosan solution is used as the casting liquid. The solution is pulsed through the spinning head, and the droplets fall into the alkaline coagulation bath after passing through the air bath. The core-shell structure particles are obtained by separation and washing.

2. The core-shell structured particle according to claim 1, characterized in that: The length of the air bath is 10 to 20 cm.

3. The core-shell structured particle according to claim 1, characterized in that: The mass concentration of the trehalose solution is 30-45%.

4. The core-shell structured particle according to claim 1, characterized in that: The mass concentration of the ganoderma lucidum source chitosan solution is 2-5%.

5. The core-shell structured particle according to claim 1, characterized in that: The mass ratio of the trehalose solution to the chitosan solution is 1:(3-8).

6. The core-shell structured particle according to claim 1, characterized in that: The trehalose solution also contains molecules that have a repairing effect on the wound surface.

7. The core-shell structured particle according to claim 1, characterized in that: The pH of the trehalose solution is 5.0-6.

8.

8. The core-shell structured particle according to claim 1, characterized in that: The coagulation bath is a NaOH solution or an ethanol solution added with NaOH.

9. A dressing for promoting wound healing, characterized in that: The core-shell structured particles according to any one of claims 1 to 8 are added thereto.

10. The dressing according to claim 9, characterized in that The dressing is a hydrogel, a spray, a patch or an ointment.

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