CN103083721B - Artificial skin graft for genipin immobilized activin B and preparation method thereof - Google Patents
Artificial skin graft for genipin immobilized activin B and preparation method thereof Download PDFInfo
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- CN103083721B CN103083721B CN201310020874.9A CN201310020874A CN103083721B CN 103083721 B CN103083721 B CN 103083721B CN 201310020874 A CN201310020874 A CN 201310020874A CN 103083721 B CN103083721 B CN 103083721B
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Abstract
The invention discloses an artificial skin graft for genipin immobilized activin B and a preparation method thereof. The artificial skin graft comprises hydrogel prepared from a skin graft stent material, and activin B which is distributed in the hydrogel and is cross-linked through genipin. The preparation method comprises the following steps of: sequentially adding an aqueous solution of genipin and the activin B into an aqueous solution of the skin graft stent material, uniformly mixing, forming, processing, gelatinizing and obtaining the product. The product is non-toxic, does not have pyrogenicity and has a high biocompatibility; pathogenic microorganism intrusion can be prevented, and infection incidence is reduced; quick healing of epithelial tissues can be promoted; and moreover, the exudation of wound surface fluid is reduced, a locally wet microenvironment is kept, the conventional disinfection and sterilization can be performed, and the artificial skin graft is convenient to storage and transport and is applied to burn, trauma and other skin injury repair.
Description
Technical field
The present invention relates to a kind of artificial skin transplant and preparation method, relate in particular to a kind of genipin fixedly artificial skin transplant and the preparation method of activin B somatomedin.
Background technology
Along with the continuous progress of society and developing rapidly of medical science, many diseases are controlled effectively, but skin trauma is growing on and on.At present, the whole nation has millions of people injured by it, therefore, skin trauma is very important disease, as burn or wound, these can damage epidermal area and the skin corium of skin, even cause holostrome skin damage, when treating the wound that cannot heal, often need carry out permanent closure wound by auto-skin grafting, and for large-area burn wound or patient's wound regeneration capacity compared with for weak person, can be limited for the position of bark fetching; For open infected wound, utilize thin parting skin graft to make the success rate of skin transplantation treatment very high, but, because the thin skin corium that splits skin graft is thinner, wound can form eventually the cicatrix contractura phenomenon of hypertrophy or contraction and affect rebuild after skin function.Therefore, various biological dressing is suggested, and the shown substitute that can be used as parting skin graft.
Along with going deep into of researching wound healing, people recognize and use the object of biological dressing not just for flap coverage, also must can help wound healing, prevent bacteria attack, reduce superelevation metabolism and the malnutrition of wound area, reduce pain and discomfort, prevent incrustation and promotion healing.
The biological dressing of clinical use amount maximum is medical absorbent cotton, because granulation tissue causes adhesion to growth in dressing, produce misery and secondary insult, and hydrops easily causes infection when removing.For this situation, many new pattern compress had been there are in recent years, what be well received by the public is that macromolecule hydrogel skin trauma is repaired dressing, it has fine air permeability and water penetration, can resist the intrusion of antibacterial, and can be according to the profile of wound surface and molding, its weak point is colloid intensity difference, lacks ruggedness.In order to increase colloid intensity, macromolecule hydrogel skin trauma is repaired in the building-up process of dressing, conventionally adopt chemical crosslinking, physics freeze thawing, three kinds of methods of radiation synthesis, adopt chemosynthesis need to use the toxic and harmful substances such as cross-linking agent and initiator, as glutaraldehyde, carbodiimides, when this makes again synthetic aerogel dressing for wound care, increased application risk; Adopt freeze-thaw method synthetic water gel dressing not have use toxic and harmful substance, its cytotoxicity is increased, have potential safety problem.But synthetic dressing has thermal reversibility, when temperature is higher, be easily out of shape, making it to be difficult for transportation stores, and frozen-thaw process easily makes the moisture in aerogel dressing overflow, cause its water holding capacity greatly to reduce, the ability of absorbing wound exudate also reduces, thereby affects its wound care effect, is difficult for laminating with skin.Adopt the synthetic aerogel dressing of radiation method, the hydrogel mechanical strength of preparing due to radiation method is less, and while especially using single raw material irradiation, the gel of generation is more crisp.
No matter be skin graft, or the substitute of skin graft " biological dressing ", for promoting wound healing, and nontoxic, have the substitute of the skin graft of good biocompatibility, be still the mankind in the urgent need to the direction of active research.
Summary of the invention
The object of the present invention is to provide a kind of genipin fixedly artificial skin transplant and the preparation method of activin B.
The technical solution used in the present invention is:
Genipin is an artificial skin transplant for activin B fixedly, comprises the hydrogel of being made by skin graft timbering material, and be distributed in hydrogel through the crosslinked activin B of genipin.
Preferably, skin graft timbering material is at least one in chitosan, carboxymethyl chitosan, alginate, gelatin and collagen protein.
Preferably, skin graft timbering material is gelatin and carboxymethyl chitosan.
Preferably, in the aqueous solution of skin graft timbering material, add successively genipin aqueous solution, activin B, mix homogeneously, obtains mixed liquor, processing and forming then, and plastic, obtains the fixedly artificial skin transplant of activin B of genipin simultaneously.
Preferably, in mixed liquor, the concentration of skin graft timbering material is 5～20 g/mL.
Preferably, skin graft timbering material is that mass ratio is the gelatin of 9:1, the mixture of carboxymethyl chitosan.
Preferably, in mixed liquor, the concentration of activin B is 1～20ng/mL.
Preferably, in mixed liquor, the concentration of genipin is 0.01～0.5 g/mL.
Preferably, in preparation process, each raw material is aseptic raw material.
The invention has the beneficial effects as follows:
Artificial skin transplant of the present invention is nontoxic, without pyrogenicity, there is good biocompatibility; Can prevent pathogenic microorganism invasion, reduce infection rate; Can promote epithelial tissue quickly-healing; Reduce wound surface body fluid and ooze out, and keep local moistening microenvironment, can routine disinfection and sterilizing, be convenient to store and transportation, be applied in the skin trauma reparations such as burn, wound.
Accompanying drawing explanation
Fig. 1 is the artificial skin transplant figure of the different gelatin of embodiment gained, chitosan content.
The specific embodiment
Below in conjunction with specific embodiment, the present invention is further illustrated, but be not limited to this.
embodiment 1
Genipin is a preparation method for the artificial skin transplant of activin B fixedly, comprises the following steps:
1) feedstock purification:
Take 5g gelatin and be dissolved in 100mL distilled water, be heated to 50 ℃ of stirring and dissolving;
Take 10g carboxymethyl chitosan and be dissolved in 100mL distilled water, stir under room temperature and dissolve;
By the gelatin after dissolving, carboxymethyl chitosan sugar juice 15000 turn/min high speed centrifugation 5min respectively, separation obtains supernatant, and lyophilization 24h, obtains respectively gelatin, carboxymethyl chitosan after purification respectively;
2) the fixedly artificial skin transplant preparation of activin B somatomedin of genipin
Take respectively gelatin 2.25g, carboxymethyl chitosan 0.25g after purification, ultraviolet sterilization 30min, then all joins in the distilled water of sterilizing, is heated to 50 ℃ of stirring and dissolving and is cooled to room temperature, with the distilled water of sterilizing, be settled to 40mL, obtain gelatin, carboxymethyl chitosan mixed liquor;
Take in the distilled water that 25mg genipin is dissolved in 10mL sterilizing, filtration sterilization, obtains genipin aqueous solution;
Pour genipin aqueous solution into gelatin, in carboxymethyl chitosan mixed liquor, under room temperature, stir evenly, activin B solution (the R & D Systems that adds 50 μ L 10 μ g/mL, Inc. Catalog Number:659-AB/CF), after stirring evenly, pour in mould, be placed in plastic at 4 ℃, obtain the fixedly artificial skin transplant of activin B somatomedin of genipin, products obtained therefrom is as for 4 ℃ or-70 ℃ of preservations, the final ratio of aqueous lower each composition of product is: gelatin 4.5 g/mL, carboxymethyl chitosan 0.5 g/mL, genipin 0.05 g/mL, activin B 10ng/mL.The left figure in Fig. 1 is shown in by the schematic diagram of products obtained therefrom.
embodiment 2
Genipin is a preparation method for the artificial skin transplant of activin B fixedly, comprises the following steps:
1) method of pressing embodiment 1 is purified gelatin, carboxymethyl chitosan;
2) preparation of artificial skin transplant:
Take respectively gelatin 4.5g, carboxymethyl chitosan 0.5g after purification, ultraviolet sterilization 30min, then joins in the distilled water of sterilizing, is heated to 50 ℃ of stirring and dissolving and is cooled to room temperature, with the distilled water of sterilizing, be settled to 40mL, obtain gelatin, carboxymethyl chitosan mixed liquor;
Take in the distilled water that 25mg genipin is dissolved in 10mL sterilizing, filtration sterilization, obtains genipin aqueous solution;
Genipin aqueous solution is poured in gelatin, carboxymethyl chitosan mixed liquor, under room temperature, stir evenly, add the activin B solution of 50 μ L 10 μ g/mL, after stirring evenly, pour in mould, be placed in plastic at 4 ℃, obtain the fixedly artificial skin transplant of activin B somatomedin of genipin, products obtained therefrom is as for 4 ℃ or-70 ℃ of preservations, and the final ratio of aqueous lower each composition of product is: gelatin 9 g/mL, carboxymethyl chitosan 1 g/mL, genipin 0.05 g/mL, activin B 10ng/mL.The middle figure in Fig. 1 is shown in by the schematic diagram of products obtained therefrom.
embodiment 3
Genipin is a preparation method for the artificial skin transplant of activin B fixedly, comprises the following steps:
1) method of pressing embodiment 1 is purified gelatin, carboxymethyl chitosan;
2) preparation of artificial skin transplant:
Take respectively gelatin 9g, carboxymethyl chitosan 1g after purification, ultraviolet sterilization 30min, then all joins in the distilled water of sterilizing, is heated to 50 ℃ of stirring and dissolving and is cooled to room temperature, with the distilled water of sterilizing, be settled to 40mL, obtain gelatin, carboxymethyl chitosan mixed liquor;
Take in the distilled water that 25mg genipin is dissolved in 10mL sterilizing, filtration sterilization, obtains genipin aqueous solution;
Genipin aqueous solution is poured in gelatin, carboxymethyl chitosan mixed liquor, under room temperature, stir evenly, add the activin B solution of 50 μ L 10 μ g/mL, after stirring evenly, pour in mould, be placed in plastic at 4 ℃, obtain the fixedly artificial skin transplant of activin B somatomedin of genipin, products obtained therefrom is as for 4 ℃ or-70 ℃ of preservations, and the final ratio of aqueous lower each composition of product is: gelatin 18g/mL, carboxymethyl chitosan 2g/mL, genipin 0.05 g/mL, activin B 10ng/mL.The right figure in Fig. 1 is shown in by the schematic diagram of products obtained therefrom.Three products from left to right in Fig. 1, color raises gradually, transparency also reduces gradually, the intensity of product from left to right also raises successively, the total content of visible gelatin, carboxymethyl chitosan is lower, and gained artificial skin transplant is more soft, and hardness is less, according to different diseases, select the artificial skin transplant of varying strength.
embodiment 4
Genipin is a preparation method for the artificial skin transplant of activin B fixedly, comprises the following steps:
1) method of pressing embodiment 1 is purified gelatin, carboxymethyl chitosan;
2) preparation of artificial skin transplant:
Take respectively gelatin 5g, carboxymethyl chitosan 1g after purification, ultraviolet sterilization 30min, then all joins in the distilled water of sterilizing, is heated to 50 ℃ of stirring and dissolving and is cooled to room temperature, with the distilled water of sterilizing, be settled to 40mL, obtain gelatin, carboxymethyl chitosan mixed liquor;
Take in the distilled water that 5mg genipin is dissolved in 10mL sterilizing, filtration sterilization, obtains genipin aqueous solution;
Genipin aqueous solution is poured in gelatin, carboxymethyl chitosan mixed liquor, under room temperature, stir evenly, the activin B solution that adds 5 μ L 10 μ g/mL, after stirring evenly, pour in mould, be placed in plastic at 4 ℃, obtain the fixedly artificial skin transplant of activin B somatomedin of genipin, products obtained therefrom is as for 4 ℃ or-70 ℃ of preservations.The final ratio of aqueous lower each composition of product is: gelatin 10 g/mL, carboxymethyl chitosan 2 g/mL, genipin 0.01 g/mL, activin B 1ng/mL.
embodiment 5
Genipin is a preparation method for the artificial skin transplant of activin B fixedly, comprises the following steps:
1) method of pressing embodiment 1 is purified gelatin, carboxymethyl chitosan;
2) preparation of artificial skin transplant:
Take respectively gelatin 5g, carboxymethyl chitosan 1g after purification, ultraviolet sterilization 30min, then all joins in the distilled water of sterilizing, is heated to 50 ℃ of stirring and dissolving and is cooled to room temperature, with the distilled water of sterilizing, be settled to 40mL, obtain gelatin, carboxymethyl chitosan mixed liquor;
Take in the distilled water that 0.25mg genipin is dissolved in 10mL sterilizing, filtration sterilization, obtains genipin aqueous solution;
Genipin aqueous solution is poured in gelatin, carboxymethyl chitosan mixed liquor, under room temperature, stir evenly, the activin B solution that adds 100 μ L 10 μ g/mL, after stirring evenly, pour in mould, be placed in plastic at 4 ℃, obtain the fixedly artificial skin transplant of activin B somatomedin of genipin, products obtained therefrom is as for 4 ℃ or-70 ℃ of preservations.The final ratio of aqueous lower each composition of product is: gelatin 10 g/mL, carboxymethyl chitosan 2 g/mL, genipin 0.5 g/mL, activin B 20ng/mL.
Claims (3)
1. the fixing artificial skin transplant for activin B of genipin, comprises the hydrogel of being made by skin graft timbering material, and be distributed in hydrogel through the crosslinked activin B of genipin; Skin graft timbering material is gelatin and carboxymethyl chitosan; Its preparation method comprises the following steps: in the aqueous solution of skin graft timbering material, add successively genipin aqueous solution, activin B, mix homogeneously, obtain mixed liquor, then processing and forming, plastic, obtains the fixedly artificial skin transplant of activin B of genipin simultaneously;
Wherein, in mixed liquor, the concentration of skin graft timbering material is 5～20 g/mL;
In mixed liquor, the concentration of activin B is 1～20ng/mL;
In mixed liquor, the concentration of genipin is 0.01～0.5 g/mL.
2. the preparation method of artificial skin transplant according to claim 1, is characterized in that: skin graft timbering material is that mass ratio is the gelatin of 9:1, the mixture of carboxymethyl chitosan.
3. the preparation method of artificial skin transplant according to claim 1, is characterized in that: in preparation process, each raw material is aseptic raw material.
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| CN103721293A (en) * | 2013-07-25 | 2014-04-16 | 天津大学 | Photo-crosslinking multilayer gradient hydrogel capable of controllably releasing active factors and preparation method of hydrogel |
| CN106496598B (en) * | 2015-09-30 | 2019-04-30 | 四川大学 | Aquagel of high bioactivity and preparation method thereof and purposes |
| CN115404199B (en) * | 2022-11-01 | 2023-03-21 | 广州优特佳生物科技发展有限公司 | Application of activin B in promoting hyaluronic acid synthesis |
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| WO2008100534A2 (en) * | 2007-02-12 | 2008-08-21 | Trustees Of Columbia University In The City Of New York | Biomimetic nanofiber scaffold for soft tissue and soft tissue-to-bone repair, augmentation and replacement |
| US20090117166A1 (en) * | 2007-08-15 | 2009-05-07 | David Myung | Sequential coupling of biomolecule layers to polymers |
| CN101693122B (en) * | 2009-10-22 | 2012-12-12 | 浙江大学 | Method for preparing high molecular degradable skin dressing and application |
| CN102552880A (en) * | 2011-12-14 | 2012-07-11 | 南方医科大学 | Biological preparation for promoting skin wound to be healed |
| CN102698313B (en) * | 2012-01-11 | 2014-07-16 | 北京大学 | Nano-silver antibacterial hydrogel and preparation method thereof |
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Effective date of registration: 20221201 Address after: Room B13, Room 401, No. 1, Kehui 1st Street, Huangpu District, Guangzhou, Guangdong 510,000 Patentee after: Guangzhou Youtejia Biotechnology Development Co.,Ltd. Address before: 510515, No. 1023 Sha Sha Road, Baiyun District, Guangdong, Guangzhou Patentee before: SOUTHERN MEDICAL University |
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