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Preparation method and application of artificial microparticle skin

A micro-skin and artificial technology, applied in medical science, bandages, prostheses, etc., can solve the problem of not having a complete skin structure, achieve the effect of improving the quality of wound healing, simplifying the treatment method, and excellent decellularization effect

Pending Publication Date: 2021-03-12
THE FIRST AFFILIATED HOSPITAL OF ARMY MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It can only function as a biological dressing in the process of repairing wounds, and does not have the function of forming a complete skin structure

Method used

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  • Preparation method and application of artificial microparticle skin
  • Preparation method and application of artificial microparticle skin
  • Preparation method and application of artificial microparticle skin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Embodiment one, a kind of preparation method of artificial particle skin, it comprises the steps:

[0050] S1, Preparation of acellular dermal matrix microparticles.

[0051] S11, soak the pigskin tissue in 75% ethanol for 1 min, wash it twice with PBS solution, scrape off excess body hair, and trim the subcutaneous adipose tissue with surgical scissors; then use 0.25% neutral protease at 4°C After digestion for 12 hours, the epidermis and dermis were separated, and the epidermis was removed to retain the dermis.

[0052] S12, pulverizing the dermal tissue into particles and adding EDTA-trypsin solution to obtain a suspension, the concentration of EDTA in the EDTA-trypsin solution is 0.02%, and the concentration of trypsin is 0.25%. The EDTA-trypsin solution with dermal tissue particles was placed in a constant temperature shaker, and shaken at a speed of 80 rpm for 24 hours, and the solution was changed every 6 hours. Then washed twice with PBS solution, ultrasonical...

Embodiment 2

[0070] The second embodiment analyzes the effect of the artificial microparticle skin prepared in the first embodiment on deep wound healing after burns, which specifically includes the following steps.

[0071] Step 1: Take 15 7-week-old nude mice grown in the same environment, divide male and female into 3 groups, 5 mice in each group, and name them as ADM group, EpSC group, and artificial microskin group respectively, and number them for adaptive feeding. Used for experiment after 1 week.

[0072] Step 2: Use 2.4% chloral hydrate solution (7.5mL / kg) for intraperitoneal injection for anesthesia. After disinfection, the puncher will make holes symmetrically on the central axis of the back skin of nude mice, causing full-thickness skin defect wounds. Each has a circular hole with a diameter of 6mm, one side is used as the control surface, and the other side is used as the experimental surface;

[0073] Step 3: Collect the artificial microparticle skin prepared in Example 1 in...

Embodiment 3

[0083] Embodiment three, a kind of preparation method of artificial particle skin, it comprises the steps:

[0084] S1, Preparation of acellular dermal matrix microparticles.

[0085] S11, soak the pigskin tissue in 75% ethanol for 1 min, wash it twice with PBS solution, scrape off excess body hair, and trim the subcutaneous adipose tissue with surgical scissors; then use 0.25% neutral protease at 4°C After digestion for 12 hours, the epidermis and dermis were separated, and the epidermis was removed to retain the dermis.

[0086] S12, pulverize the dermal tissue into particles and add EDTA-trypsin solution to obtain a suspension. The concentration of EDTA in the EDTA-trypsin solution is 0.01%, and the concentration of trypsin is 0.5%. The EDTA-trypsin solution with dermal tissue particles was placed in a constant temperature shaker, and shaken at a speed of 80 rpm for 24 hours, and the solution was changed every 6 hours. Then washed twice with PBS solution, ultrasonically o...

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Abstract

The invention discloses a preparation method of artificial microparticle skin. The preparation method comprises the following steps: S1, preparing acellular dermal matrix microparticles; S2, performing epidermal stem cell isolated culturing; S3, combining the acellular dermal matrix particles with epidermal stem cells; S31, soaking and coating the acellular dermal matrix particles prepared in thestep S1 with a IV-type collagen solution at the temperature of 4-8 DEG C, and performing reheating to room temperature to obtain an acellular dermal matrix particle suspension; S32, dropwise adding the acellular dermal matrix particle suspension into a cell culture plate, and resuspending acellular dermal matrix particles in the cell culture plate by adopting a KSFM or Epilife culture solution; S33, digesting and collecting the epidermal stem cells cultured in the S2 in the logarithmic phase, adjusting the concentration to be (0.2-0.5) * 10 <6> cells / ml, adding the epidermal stem cells into acell culture plate, and performing culturing to obtain the artificial microdermis. The fusion of the dermal matrix and the epidermal stem cells can be realized, and the prepared artificial microparticle skin has the effects of quickly promoting wound healing and improving the wound healing quality.

Description

technical field [0001] The invention relates to the field of tissue engineering biomaterials, in particular to a preparation method and application of artificial particle skins. Background technique [0002] The repair of severe burn wounds has always been a difficult point in the research of burn medicine, and the lack of skin source is the fundamental problem. The current clinical application of micro-skin grafting, MEEK micro-skin grafting, and autologous skin mosaic grafting can meet the needs of 1:14 wound closure (1% of the skin area can be expanded to cover 14% of the wound surface). However, this traditional treatment strategy of "removing the east wall to make up the west wall" needs to sacrifice a large amount of normal skin, and severe burns often require multiple operations to seal the wound, which takes a long time for intensive care and high life risk. In addition, there have been studies on the use of skin layer fibroblasts, umbilical cord mesenchymal stem ce...

Claims

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Application Information

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IPC IPC(8): A61L27/36A61L27/38A61L27/60A61L15/40A61L15/46
CPCA61L27/362A61L27/3633A61L27/3834A61L27/3687A61L27/3691A61L15/40A61L15/46A61L27/60A61L2300/412
Inventor 詹日兴赵小红罗高兴郭熠城
Owner THE FIRST AFFILIATED HOSPITAL OF ARMY MEDICAL UNIV
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