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Diabetes ulcer repair material and method for preparing same

A technology for diabetic ulcers and repairing materials, applied in the field of repairing materials for diabetic ulcers and their preparation, can solve the problems of complicated material preparation process, easy infection of wounds, limited cell sources, etc., achieves good biocompatibility, promotes Wound healing, wide-ranging effects

Active Publication Date: 2019-03-12
WEST CHINA HOSPITAL SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It overcomes the shortcomings of the existing diabetic ulcer repair materials, such as limited cell sources, invasive acquisition, difficulty in survival, complicated material preparation process, secondary trauma caused by acquisition, strong immunogenicity, and prone to infection on the wound surface

Method used

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  • Diabetes ulcer repair material and method for preparing same
  • Diabetes ulcer repair material and method for preparing same
  • Diabetes ulcer repair material and method for preparing same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0178] Example 1 A method for preparing a repair material modified with extracellular matrix for diabetic ulcers

[0179] 1. Primary culture and cryopreservation of USCs

[0180] (1) Primary culture: take 50-250mL of urine from healthy ordinary people, put it into 50mL centrifuge tubes, centrifuge at 400g for 10min in a centrifuge, discard the upper layer of urine on a clean bench, add 40mL containing 1 % double antibody in PBS, mixed with the bottom urine, and centrifuged at 400g for 5min in a centrifuge. Repeat the step of washing the urine with PBS twice. Discard the supernatant, add 5mL USCs complete medium to resuspend the cells at the bottom of the centrifuge tube, then pipette the cell suspension into a T25 culture flask, and place it at 37°C containing 5% CO 2 Cultivate in the incubator, and observe whether there is pollution regularly in the first two days; replace the medium after cell clones appear.

[0181](2) Cryopreservation: USCs cells grown to about 70%-80% ...

Embodiment 2

[0199] Example 2 A method for preparing a repair material modified with extracellular matrix for diabetic ulcers

[0200] 1. Primary culture and cryopreservation of USCs

[0201] Same as Example 1.

[0202] 2. Preparation of SIS

[0203] Same as Example 1.

[0204] 3. Cell inoculation

[0205] Same as Example 1.

[0206] 4. Decellularization of USCs-SIS composites

[0207] Triton X-100 combined with DNase I treatment: the SIS co-cultured with USCs for 7 days was taken out, washed 3 times with PBS, 2 min each time, the USCs-SIS composite material was placed on a shaker and washed with 0.5% Triton X-100 solution for 5 min, Then place the USCs-SIS composite material on a shaker and rinse with 100 U / mL DNase I solution for 5 minutes, and finally wash with deionized water for 5 minutes. The above steps are repeated 3 times, and the last wash is 30 minutes. , namely the extracellular matrix-modified repair material (D-USCs-SIS).

experiment example 1

[0218] Experimental Example 1 Identification of USCs

[0219] (1) Morphological characteristics of USCs

[0220] 1. Experimental method

[0221] During the primary culture of USCs, multiple centrifugation methods are used. About 3 to 14 days after inoculation, individual cells can be observed to adhere to the wall, usually 1 to 3 cells adhere to the wall, and the cells that do not adhere to the wall are suspended in the medium. The adhered cells continued to proliferate and formed densely arranged cell colonies in about 10 days. When the cells are fused to 80%, the cells are subcultured with TNE at a ratio of 1:3 / 4. After about 3 to 4 days, the cells reach a state of complete confluence, and can continue to be subcultured. After the USCs passed to the third generation, their morphological characteristics were observed by optical microscope.

[0222] 2. Experimental results

[0223] The morphological characteristics of USCs after passing to the third generation are as follo...

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Abstract

The invention discloses a diabetes ulcer repair material modified by extracellular matrixes. The diabetes ulcer repair material is a small intestine submucosa membrane modified by the extracellular matrixes of human urine-derived stem cells. The diabetes ulcer repair material has the advantages that the diabetes ulcer repair material is excellent in biocompatibility and bioactivity, the immunogenicity can be removed to the greatest extent, and the durations of inflammatory periods can be shortened; the extracellular matrixes and growth factors in the matrixes can be retained, diabetes skin ulcer wound healing can be promoted, the cells and materials for the diabetes ulcer repair material can come from extensive sources, and secondary injury on patients can be prevented in preparation procedures; the shortcomings of limited cell sources for existing diabetes ulcer repair materials, difficulty in survival, complicated material preparation procedures, secondary trauma, high immunogenicityand susceptibility to infection of wound can be overcome by the aid of the diabetes ulcer repair material, and accordingly the diabetes ulcer repair material has an excellent clinical application prospect.

Description

technical field [0001] The invention relates to an extracellular matrix-modified repair material for diabetic ulcers and a preparation method thereof. Background technique [0002] In recent years, the incidence of diabetes has gradually increased. As of 2015, 1 in every 11 adults worldwide has diabetes (a total of 415 million); there are 318 million pre-diabetic patients, and the prevalence of diabetes is as high as 8.8%. , the prevalence of prediabetes was 6.7%. Diabetic skin ulcers are a refractory complication of diabetes, and the probability of developing ulcers in each diabetic patient's life cycle is as high as 15%. Diabetic skin ulcer wounds are susceptible to infection, inactivated or abnormally distributed growth factors, decreased local blood flow, and hypoxia, etc., resulting in prolonged wound healing time, or even non-healing resulting in disability or amputation. In response to this complication, departments such as burns and plastic surgery, dermatology, or...

Claims

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Application Information

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IPC IPC(8): A61L27/60A61L27/36A61L27/50A61L27/38
CPCA61L27/3629A61L27/3633A61L27/3687A61L27/3834A61L27/3895A61L27/50A61L27/60A61L2430/40
Inventor 解慧琪汪祝乐黄益洲
Owner WEST CHINA HOSPITAL SICHUAN UNIV
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