I-type recombinant collagen gel dressing as well as preparation method and application thereof

Abstract

The invention discloses an I-type recombinant collagen gel dressing as well as a preparation method and application thereof. The dressing comprises the following main components: sodium alginate, I-type recombinant collagen and methylparaben. The preparation method comprises the following steps: sequentially or simultaneously weighing the main components in parts by weight, adding a weakly acidic phosphate buffer solution, stirring and dissolving by using a stirrer to form I-type recombinant collagen gel, centrifugally defoaming, sterilizing, filling and storing at normal temperature. The I-type recombinant collagen gel dressing has the advantages of simple and easily available raw materials, few steps of the preparation method, simple operation, no hormone and heavy metal, no toxicity and no irritation, can be directly taken and coated on a wound surface, and can also be ground into powder for use after freeze drying, or the powder is taken and processed into a solid filler for use, so that the wound is promoted to be quickly healed and repaired within 20 days, and the wound healing effect is good. The generation of scars is inhibited. The gel dressing can be applied to various skin wounds or in-vivo mucous membrane surfaces.

Description

Technical Field

[0001] The present invention relates to the technical field of biomedical surgical dressings, and particularly relates to a type I recombinant collagen gel dressing, a preparation method thereof, and an application thereof. Background Art

[0002] The skin covers the body surface and is one of the largest organs of the human body. The skin is the first barrier between the body and nature, and has protective functions, absorption functions, sensory functions, body temperature regulation functions, metabolic functions, and immune functions.

[0003] A skin wound is skin damage caused by external injury factors such as surgical operations, external forces, heat, electric current, chemical substances, low temperature, and internal factors of the body such as local blood supply disorders. At this time, the barrier function of the skin is damaged, and pathogens are likely to invade the body through the skin wound, causing further damage. Therefore, it is urgent to heal the wound to restore the barrier function. According to the healing cycle of skin wounds, they can be divided into acute wounds and chronic wounds. Wound healing refers to a series of pathophysiological processes in which local tissues are repaired through regeneration, repair, and reconstruction after tissue loss caused by the action of injury factors. The healing process mainly includes a coagulation phase, an inflammation phase, a repair phase, and a maturation phase.

[0004] In order to promote the rapid healing of skin wounds, various dressings are commonly used for assistance. These dressings can be divided into traditional dressings, synthetic dressings, and biological dressings according to their different materials. Traditional dressings are mostly inert dressings in a solid state, including absorbent cotton, gauze, bandages, etc., which have no promoting effect on wound healing and only have a physical protection effect. Such dressings are prone to adhering to the wound surface and are likely to cause secondary mechanical damage to the wound during dressing change. Synthetic dressings and biological dressings mainly include hydrogel dressings, hydrocolloid dressings, etc., which have the advantages of good air permeability, high water content, easy absorption, etc. However, the frequent dressing changes of such dressings result in high usage costs, and dressing debris is likely to remain in the wound during dressing change, leading to scar formation during the wound healing process.

[0005] Chinese Patent with publication number CN109568653A discloses a recombinant human collagen gel dressing for wounds, a preparation method thereof, and a using method thereof. Among them, the pH of the hydrogel dressing is maintained between 7.3 and 7.5, which is weakly alkaline. Compared with the weakly acidic state of normal skin, there are certain biocompatibility problems, and pigment deposition can be observed on the surface of the healed wound, and it cannot be completely restored to the skin state before being damaged, leaving a mark.

[0006] Therefore, it is an urgent problem to be solved in the field to produce wound dressings that are inexpensive, effective, have good biocompatibility, can inhibit scar formation, and have good recovery effects. Summary of the Invention

[0007] The object of the present invention is to provide a type I recombinant collagen gel dressing, its preparation method and application.

[0008] In the first aspect of the present invention, there is provided a type I recombinant collagen gel dressing, the main components of which include sodium alginate, type I recombinant collagen, methylparaben and phosphate buffer solution.

[0009] Preferably, the mass concentrations of the main components of the type I recombinant collagen gel dressing are: sodium alginate 3.9 - 21.3 g / L, type I recombinant collagen 0.3 - 1.8 g / L, methylparaben 0.09 - 0.44 g / L, and phosphate buffer solution as the solvent.

[0010] Preferably, the pH value range of the phosphate buffer solution is 5.7 - 7.0.

[0011] In the second aspect of the present invention, there is provided a preparation method of a type I recombinant collagen gel dressing, which comprises the following steps: S1. Prepare phosphate buffer solution by mixing sodium dihydrogen phosphate and disodium hydrogen phosphate in proportion; S2. Weigh sodium alginate, type I recombinant collagen, and methylparaben in sequence or simultaneously and place them in a stirring container; S3. Add the phosphate buffer solution prepared in S1 to the target mass concentration, stir and dissolve to form a gel; S4. Centrifuge to remove bubbles; S5. After sterilization, fill and store at room temperature; Preferably, in S2, the mass of each main component contained in every 1000 ml of the dressing is: sodium alginate 3.9 - 21.3 g, type I recombinant collagen 0.3 - 1.8 g, methylparaben 0.09 - 0.44 g.

[0012] Preferably, in S3, the stirring speed for stirring and dissolving with a stirrer is 500 - 3000 r / min, and the stirring time is 0.3 - 5 h.

[0013] Preferably, in S4, the speed of centrifuging to remove bubbles is 1000 - 3000 r / min, and the time is 30 min - 2 h.

[0014] Preferably, in S5, the sterilization method is ultraviolet irradiation sterilization or ethylene oxide sterilization; the filling capacity can be selected as 2 ml, 5 ml, 10 ml, 20 ml and 50 ml according to requirements.

[0015] In the third aspect of the present invention, there is provided the application of the type I recombinant collagen gel dressing in repairing skin wounds and internal surface mucosae at least in daily life or medicine.

[0016] Preferably, the skin wound surface includes surgical wounds, burns, scalds, lacerations, and diabetic foot ulcers; the surface of the internal mucous membrane includes the surface of female reproductive organs, oral mucous membrane, nasal mucous membrane, and perianal tissue surface.

[0017] Preferably, the gel dressing can be freeze-dried and ground into powder for use, or the powder can be made into a solid filler for use. The solid filler mainly includes sterile fillers required during treatment, such as filling dental cavities.

[0018] For the surgical wound surface such as the cesarean section suture wound surface, under the action of the gel dressing, its components provide a moist healing environment for the wound, optimize cell growth, accelerate cell proliferation, reduce the inflammatory response, promote wound healing, and inhibit scar formation.

[0019] The burns, scalds, and lacerations mainly include small wounds commonly seen in people's daily lives, which can heal on their own by using the gel dressing without other medical interventions in the short term. For severe burns, scalds, or serious limb fractures, the gel dressing described in this application can be used as an auxiliary means and cannot completely heal the wound without relying on other medications.

[0020] The diabetic foot refers to a disease state in which the feet of diabetic patients have reduced lower limb protection function due to neuropathy, and large and small blood vessel lesions cause insufficient arterial perfusion, resulting in microcirculation disorders and ulcers and gangrene. Diabetic foot is a serious complication of diabetes and one of the important reasons for disability and even death in diabetic patients, causing pain to patients. The gel dressing provided in this application can improve the condition of diabetic foot and relieve the pain of patients.

[0021] The internal mucous membrane includes the mucous membranes in the human body that have a certain contact with the outside world, such as the vaginal wall and cervix wall of female reproductive organs, oral mucous membrane, nasal mucous membrane, and perianal tissue surface. Due to its aseptic, sufficient moisture, and non-specific shape characteristics, the gel dressing can be made into a suppository for administration, reducing the friction and pain caused during the dressing change process in sensitive areas such as the private parts and perianal area of patients, and accelerating the healing rate of wounds or inflammation.

[0022] Compared with the prior art, the present invention has the following beneficial effects: (1) The action principle of each component: The type I recombinant collagen gel dressing provides a moist healing environment for the wound; optimizes cell growth, accelerates cell proliferation, reduces the inflammatory response, promotes wound healing and repair within 20 days, and inhibits scar formation; the gel dressing has good biocompatibility, does not contain hormones and heavy metals, and is aseptic, non-toxic, and non-irritating.

[0023] (2) Simple production process: The raw materials are easily obtainable, the method steps are few, the operation is simple, the production cost is controllable, the product is convenient to store, the product can be further made into various forms such as powders and solid fillers, and the aseptic process is suitable for large-scale production in the medical industry, and it can be stored at room temperature, reducing storage and transportation costs.

[0024] (3) Wide range of application fields: The gel dressing provided in this application can meet the needs of a variety of users: on the one hand, due to its aseptic design, it can be used as the raw material for the production of Class II medical devices, and the gel dressing is suitable for skin wounds or the surface of internal mucous membranes; on the other hand, the gel dressing can be freeze-dried and ground into powder for use, or further processed into solid fillers for use, meeting the treatment needs of daily small wounds. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 Wound healing conditions of the experimental mice in the example and the control example on the 1st, 5th, 13th and 20th days when the pH is 5.7; Figure 2 Wound healing conditions of the experimental mice in the example and the control example on the 1st, 5th, 13th and 20th days when the pH is 6.3; Figure 3 Wound healing conditions of the experimental mice in the example and the control example on the 1st, 5th, 13th and 20th days when the pH is 7.0. DETAILED DESCRIPTION OF THE INVENTION

[0026] The raw materials used in the present invention are all ordinary commercially available products, and their sources are not specifically limited. In order to enable those skilled in the art to better understand the technical solutions of the present invention, the present invention will be clearly and completely described below in conjunction with the embodiments and the drawings, and a further detailed description of the present invention will be given. Obviously, the described embodiments and the drawings are only a part of the present invention, rather than all of the embodiments and the drawings. Based on the embodiments and the drawings of the present invention, all other embodiments and drawings obtained by those of ordinary skill in the art without creative efforts shall fall within the scope of protection of the present invention.

[0027] In the examples, sodium dihydrogen phosphate and disodium hydrogen phosphate are used to prepare phosphate buffer solution according to the following steps: (1) Prepare stock solutions A and B respectively, for example: Solution A: 0.2M NaH2PO4 solution, weigh 31.2g of NaH2PO4·2H2O and dissolve it in 1000 ml of water.

[0028] Solution B: 0.2M Na2HPO4 solution, weigh 71.6g of Na2HPO4·12H2O and dissolve it in 1000 ml of water.

[0029] (2)After the preparation of Solution A and Solution B, mix Solution A and Solution B according to the ratio of the phosphate buffer solution described in Table 1 to obtain the phosphate buffer solution. Select the phosphate buffer solutions with pH = 5.7, pH = 6.3, and pH = 7.0 among them for use in the examples.

[0030] Table 1 Ratios of Solution A and Solution B of Phosphate Buffer Solution and Corresponding pH

[0031] In the examples, a skin-lesioned mouse model was prepared, and a type I recombinant collagen gel dressing was used to cover the skin wound and observe the therapeutic effect of the dressing. The specific operation steps were as follows: Select nude mice weighing 30 - 35 g. After anesthesia and observing the mice until they were in a state suitable for surgery, disinfect them with iodophor. After hair removal, use a scalpel to cut two full-thickness skin areas in the center of the back of the mice, each with a size of 0.8 - 1.5 cm 2 , to make a full-thickness skin defect wound model of mice. Clean the wound, apply the type I recombinant collagen gel dressing to one of the skin lesions as an example, and cut medical gauze into a shape suitable for the size of the mouse wound and then apply it to the mouse wound. Then take a suitable length of medical bandage to wrap the wound, paying attention to the appropriate tightness. The other skin lesion wound was used as a control example, without applying the type I recombinant collagen gel dressing, and the other treatment methods were the same as those in the example group. Example 1

[0032] Prepare type I recombinant collagen gel dressing I according to the following steps: 1. Prepare a phosphate buffer solution with pH 5.7 according to the above ratio; 2. Weigh 18.2 g of sodium alginate, 1.3 g of type I recombinant collagen, and 0.25 g of methylparaben in sequence and place them in a stirring container; 3. Add buffer solution to 1000 ml, and stir and dissolve with a stirrer at a stirring speed of 1500 r / min for 2 h; 4. Centrifuge at a rotation speed of 2000 r / min for 1.5 h to defoam; 5. After sterilization, store in cans; The therapeutic effect of Example 1 is shown in Figure 1. The wound in the example was smeared with type I recombinant collagen dressing, while the wound in the control example was not smeared with type I recombinant collagen dressing. The wound healing and scar formation of the mouse skin lesions on the first, fifth, thirteenth and twentieth days after surgery showed that on the first day after surgery, the control wound had obvious exudation, the bottom surface was all red and swollen, and only a small amount of exudation was observed in the example wound, and only the edge of the wound was red and swollen; on the fifth day after surgery, the exudation of the control group decreased, the redness and swelling were alleviated, and the redness and swelling of the example disappeared, with no obvious exudation; on the thirteenth day after surgery, the control wound was still red and swollen, and scars appeared, and the wound did not heal; the example wound was not red and swollen, and showed the pink color before the skin healed, no obvious scars appeared, and the wound area was significantly reduced; on the twentieth day after surgery, the control wound had been replaced by scar tissue, and the example wound was almost completely healed, and no scars were visible to the naked eye. Embodiment 2

[0033] Prepare type I recombinant collagen gel dressing II as follows: 1. Prepare pH 6.3 phosphate buffer according to the above ratio; 2. Weigh 18.2 g of sodium alginate, 1.3 g of type I recombinant collagen, and 0.25 g of methylparaben in sequence and place them in a stirring container; 3. Add buffer to 1000 ml and stir to dissolve with a stirrer at a speed of 1500 r / min for 2 h; 4. Centrifuge at 2000r / min for 1.5h to defoam; 5. Fill and store after sterilization; The therapeutic effect of Example 2 is shown in Figure 2. The example is a wound with gel, and the control is a wound without gel. The wound healing and scar formation of the mouse skin lesions on the first, fifth, thirteenth and twentieth days after surgery were observed. It can be seen that on the first day after surgery, the control wound had obvious exudation, redness and swelling on the bottom surface, and the example wound had a slight exudation, and the redness and swelling on the edge were not obvious; on the fifth day after surgery, the exudation of the control group decreased, the redness and swelling were alleviated, and the redness and swelling of the example disappeared, no exudation was seen, and the wound was reduced; on the thirteenth day after surgery, the control wound was still red and swollen, scars appeared, and the wound did not heal; the example wound was basically healed; on the twentieth day after surgery, the control wound was replaced by scar tissue, and the example wound was completely healed without scar formation. Embodiment 3

[0034] Prepare type I recombinant collagen gel dressing three as follows: 1. Prepare pH 7.0 phosphate buffer according to the above ratio; 2. Weigh 18.2 g of sodium alginate, 1.3 g of type I recombinant collagen, and 0.25 g of methylparaben in sequence and place them in a stirring container; 3. Add buffer to 1000 ml, stir and dissolve with a stirrer at a stirring speed of 1500 r / min for 2 h; 4. Centrifuge at a speed of 2000 r / min for 1.5 h to defoam; 5. Fill and store after sterilization; The treatment effect of Example 3 is shown in Figure 3. The example is the wound smeared with the gel, and the control example is the wound without the gel smeared. By observing the wound healing and scar formation at the first, fifth, thirteenth, and twentieth days after the operation at the skin lesion of the mouse, it can be seen that on the first day after the operation, the wound of the control example exuded significantly, the bottom surface was red and swollen, the wound of the example exuded slightly, and the wound edge was red and swollen; on the fifth day after the operation, the exudation of the control group decreased and the redness and swelling subsided, the redness and swelling of the wound in the example disappeared, and there was no obvious exudation; on the thirteenth day after the operation, the wound of the control example was still red and swollen, scars appeared, and the wound was not healed; the wound area in the example decreased significantly; on the twentieth day after the operation, the wound of the control example was replaced by scar tissue, and the wound in the example was close to healing and no scar was formed. Example 4

[0035] Prepare Type I recombinant collagen gel dressing IV according to the following steps: 1. Prepare phosphate buffer with pH 4.8 according to the above ratio; 2. Weigh 18.2 g of sodium alginate, 1.3 g of Type I recombinant collagen, and 0.25 g of methylparaben in sequence and place them in a stirring container; 3. Add buffer to 1000 ml, stir and dissolve with a stirrer at a stirring speed of 1500 r / min for 2 h; 4. Centrifuge at a speed of 2000 r / min for 1.5 h to defoam; 5. Fill and store after sterilization; Compared with Examples 1, 2, and 3, the treatment effect of Example 4 shows that the wound healing speed is significantly slowed down and a small amount of scar is formed. Example 5

[0036] Prepare Type I recombinant collagen gel dressing V according to the following steps: 1. Prepare phosphate buffer with pH 7.2 according to the above ratio; 2. Weigh 18.2 g of sodium alginate, 1.3 g of Type I recombinant collagen, and 0.25 g of methylparaben in sequence and place them in a stirring container; 3. Add buffer to 1000 ml, stir and dissolve with a stirrer at a stirring speed of 1500 r / min for 2 h; 4. Centrifuge at a speed of 2000 r / min for 1.5 h to defoam; 5. Fill and store after sterilization; The treatment effect of Example 5, compared with Examples 1, 2, and 3, showed that the wound healing rate was significantly slower, and it could not fully recover to the skin state before damage, with a small amount of scar formation.

[0037] Examples 6, 7, 8, 9, 10, and 11 show the treatment effects of type I recombinant collagen gel dressings with different mass concentrations on wounds at pH 6.3. See Table 2.

[0038]

[0039] What is not disclosed in detail in the specific implementation manner is that the type I recombinant collagen gel dressing described in the present application is at least used in daily life or medicine for repairing skin wounds and the surfaces of internal mucous membranes. The skin wounds include surgical wounds, burns, scalds, lacerations, and diabetic foot; the surfaces of internal mucous membranes include the surfaces of female reproductive organs, oral mucous membranes, nasal mucous membranes, and perianal tissues. The gel dressing can be freeze-dried, ground into powder for use, or the powder can be made into solid fillers for use. The solid fillers mainly include sterile fillers required during treatment, such as filling dental cavities, etc. This is a reasonable presumption based on the inherent characteristics of the raw materials and preparation methods of the type I recombinant collagen gel dressing provided in the present application and the prior art. At the same time, its application scope does not include the treatment of serious diseases such as severe trauma. Therefore, those skilled in the art can understand that these technical solutions can be achieved without creative labor.

[0040] It should be understood that the above examples are only for illustrating the present invention and should not be regarded as limiting the scope of the present invention. Various forms of processes shown above can be used, with steps reordered, added, or deleted. For example, the steps described in the present invention can be executed in parallel, sequentially, or in different orders, as long as the desired results of the technical solution of the present invention can be achieved, they should all be included within the protection scope of the present invention.

[0041] The above specific implementation manner does not constitute a limitation to the protection scope of the present invention. Those skilled in the art should understand that various modifications, combinations, sub-combinations, and substitutions can be made according to design requirements and other factors. All modifications, equivalent substitutions, and improvements made within the spirit and principle of the present invention should be included within the protection scope of the present invention.

Claims

1. A type I recombinant collagen gel dressing, characterized in that, The main components include sodium alginate, recombinant type I collagen, methylparaben, and phosphate buffer solution; the mass concentrations of the main components in the recombinant type I collagen gel dressing are: sodium alginate 3.9 - 21.3 g / L, recombinant type I collagen 0.3 - 1.8 g / L, methylparaben 0.09 - 0.44 g / L, and phosphate buffer solution as the solvent.

2. A preparation method of the type I recombinant collagen gel dressing according to claim 1, characterized in that, It includes the following steps: S1. Prepare phosphate buffer solution; S2. Weigh sodium alginate, recombinant type I collagen, and methylparaben by weight in sequence or simultaneously and place them in a stirring container; S3. Add the phosphate buffer solution prepared in S1 to the stirring container to the target mass concentration, stir and dissolve to form a gel; S4. Centrifuge to remove bubbles; S5. After sterilization, fill and store at room temperature.

3. The preparation method according to claim 2, wherein: In step S1, the pH range of the phosphate buffer solution is weakly acidic, 5.7 - 7.

0.

4. The preparation method according to claim 2, characterized in that: In step S3, use a stirrer to stir and dissolve, the stirring speed is 500 - 3000 r / min, and the stirring time is 0.3 - 5 h.

5. The method according to claim 2, characterized in that: In step S4, the centrifugation speed is 1000 - 3000 r / min, and the time is 0.5 - 2 h.

6. Use of the type I recombinant collagen gel dressing according to claim 1 or the type I recombinant collagen gel dressing prepared by the preparation method according to any one of claims 2-5, characterized in that This gel dressing is applied on the skin wound surface or the surface of internal mucous membranes to promote the healing and repair of the wound within 20 days.

7. The application according to claim 6, wherein This gel dressing can be used after being freeze-dried and ground into powder, or the powder can be taken to make a solid filler.

8. Use of the type I recombinant collagen gel dressing according to claim 6, characterized in that, The skin wound surface includes surgical wounds, burns, scalds, cuts, and diabetic foot.

9. Use of the type I recombinant collagen gel dressing according to claim 6, characterized in that, The surface of internal mucous membranes includes the surface of female genital organs, oral mucous membranes, nasal mucous membranes, and perianal tissues.

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