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Composition, 3D (three-dimensional) dressing with composition and preparation method thereof

A composition and 3D technology, applied in the field of 3D printing, can solve the problems of difficulty in obtaining curative effect, pain of patients, low survival rate of transplanted fat, etc., and achieve the effect of good mechanical strength

Inactive Publication Date: 2017-01-11
GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, drugs, surgery and other methods are mainly used to repair wounds clinically, and it is difficult to obtain satisfactory curative effect, which brings pain to patients.
The low survival rate of transplanted fat also greatly limits its wide application in clinic.

Method used

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  • Composition, 3D (three-dimensional) dressing with composition and preparation method thereof
  • Composition, 3D (three-dimensional) dressing with composition and preparation method thereof
  • Composition, 3D (three-dimensional) dressing with composition and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1: Preparation of human adipose acellular matrix

[0045] In the ultra-clean bench, wash the specimen 3 times with PBS buffer.

[0046] The adipose tissue was cut into 3-5mm thick tissue pieces, placed in hypotonic tris buffer (containing 10mmol / L tris, 5mmol / L EDTA), frozen at -80°C, 37 ℃ water bath rewarming, repeated freezing and thawing 5 times;

[0047] Treat with 0.25% trypsin for 6 hours, wash with sterile PBS buffer for 30 minutes, 3 times;

[0048] Treat with isopropanol for 36 hours, wash with sterile PBS buffer for 30 minutes, 3 times;

[0049] Treat with 0.25% trypsin for 6 hours, wash with sterile PBS buffer for 30 minutes, 3 times;

[0050] 5×10 7 U / L DNase-I and 1×10 6 U / L DNase-A treatment for 16 hours, washed with sterile PBS buffer for 30 minutes, 3 times;

[0051] Treat with isopropanol for 8 hours, wash with sterile PBS buffer for 30 minutes, three times.

[0052] All solutions were added with double antibodies (100U / mL penicillin, 0.1...

Embodiment 2

[0053] Example 2: Isolation and culture of human adipose-derived mesenchymal stem cells

[0054] In the ultra-clean bench, use a pipette to suck up and discard the lower layer of adipose tissue, add an equal volume of cell cleaning solution with a pipette, cover the bottle cap, shake it upside down, and let it stand until the layers are obvious; suck up and discard the lower layer of adipose tissue solution (repeat).

[0055] Use a pipette to evenly divide the adipose tissue into 50ml centrifuge tubes, add cell cleaning solution to 40ml, and centrifuge at 1500rpm for 5min. Use a pipette to discard the upper layer of fatty oil and the lower layer of liquid, and add an equal volume of 0.5% type I collagenase solution to each tube of adipose tissue. After sealing, shake it horizontally back and forth, and mix well.

[0056] Transfer to a constant temperature shaker, digest at 37°C, 200R for 30-50min, until the fat is completely digested. After the centrifuge tube is balanced, p...

Embodiment 3

[0060] Example 3: Dressing

[0061] The P3 generation adipose-derived mesenchymal stem cells were taken, the medium was sucked off, and the digestion solution was added for digestion, and then an appropriate amount of serum was used to stop the digestion, and a single-cell suspension was gently pipetted.

[0062] Centrifuge at 1500rpm for 5min, discard the supernatant, wash the cells 3 times with 4°C PBS, resuspend and mix well, and adjust the cell concentration to about 10 7 pieces / ml.

[0063] In a 50ml centrifuge tube, add chitin, type I collagen, EGF, human adipose acellular matrix freeze-dried powder, and Lonza successively according to the formula, stir thoroughly at each step, and adjust the pH value to 7.2- with 1mol / L sodium hydroxide 7.4, Form the Scaffold Material Solution.

[0064] components content P3 generation adipose stem cells (obtained in Example 1) 0.9×10 5 pcs / ml~1.1×10 5 pcs / ml

[0065] Take the single cell suspension and scaffold ...

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Abstract

The invention relates to the field of 3D (three-dimensional) printing, in particular to a composition, a 3D dressing with the composition and a preparation method thereof. The 3D dressing with the composition can be used for effectively restoring the wound surface. The technology of combining 3D printing with adipose tissue-derived stromal cells and stands is used for fast printing the 3D printing dressing used for restoring the skin wound surface. The 3D dressing has good mechanical intensity, can be clamped by operating forceps; after the addition into a culture medium, looseness cannot occur; breaking and great-amplitude degradation do not occur in the continuous culture process.

Description

technical field [0001] The invention relates to the field of 3D printing, in particular to a composition, a 3D dressing containing the composition and a preparation method thereof. Background technique [0002] Wound is the damage of normal skin (tissue) caused by external injury factors such as surgery, external force, heat, electric current, chemical substances, low temperature, and internal factors such as local blood supply disturbance. It is often accompanied by the destruction of skin integrity and the loss of a certain amount of normal tissue. At the same time, the normal function of the skin is damaged, which is also called wound or trauma. Wound repair means wound repair. [0003] The repair and healing of skin wounds is a hotspot and difficulty in surgical research, and refractory wounds such as severe burns, diabetic feet, and tumor ulcers are even more thorny issues in the field of trauma surgery. At present, medicine, surgery and other methods are mainly used t...

Claims

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Application Information

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IPC IPC(8): A61L27/24A61L27/20A61L27/36A61L27/38A61L27/54A61L26/00A61K38/18A61P17/02B33Y10/00B33Y80/00A61K35/28
CPCA61K35/28A61K38/1808A61L26/0023A61L26/0033A61L26/0057A61L26/0066A61L27/20A61L27/24A61L27/3633A61L27/3834A61L27/54A61L2300/252A61L2300/414B33Y10/00B33Y80/00C08L89/00C08L5/08
Inventor 陈海佳葛啸虎王一飞赵萌萌王小燕
Owner GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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