Biofilm capable of rapidly recovering skin wound surface, and preparation method thereof

A skin wound, rapid recovery technology, applied in biochemical equipment and methods, microorganisms, skin diseases, etc., can solve the problems of high cost, unfavorable promotion and application, slow effect, etc., to achieve low cost, reduce deterioration, and protect wounds.

Inactive Publication Date: 2017-04-26
THE THIRD AFFILIATED HOSPITAL OF GUANGZHOU MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, the commonly used treatment method is the preparation of treating diabetic wounds after culturing stem cells, but this method is slow in effect and high in cost, which is not conducive to large-scale promotion and application

Method used

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  • Biofilm capable of rapidly recovering skin wound surface, and preparation method thereof
  • Biofilm capable of rapidly recovering skin wound surface, and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1 EPC cell culture fluid calcium alginate composite biofilm (Ⅰ)

[0022] 1) Extraction of human umbilical cord blood endothelial progenitor cells: extract 60ml of umbilical cord blood from healthy newborns with a syringe, anticoagulate with sodium heparin, dilute with PBS at a volume ratio of 1:1, and slowly add Ficoll (1.077) lymphocyte separation solution containing a volume ratio of 2:1 In the centrifuge tube, use the density gradient centrifugation method, centrifuge at 2000rpm in a horizontal centrifuge for 25 minutes, and carefully absorb the mononuclear cell layer with a pipette. Then wash the cells with an equal volume of PBS, centrifuge at 2000rpm for 8 minutes, and inoculate the cells on a 25cm cell coated with human fibronectin. 2 Add an appropriate amount of endothelial cell culture medium (containing 10% fetal bovine serum) to the culture flask for culture, and culture at a saturated humidity, 5% carbon dioxide concentration, and a constant temperat...

Embodiment 2

[0027] Example 2 EPC cell culture fluid calcium alginate composite biofilm (Ⅱ)

[0028] The endothelial progenitor cells were subcultured according to the steps of 1) and 2) in Example 1. After culturing the cells to the third generation, when the cells grow to 70-80% confluence, place the cells in 0.5% oxygen concentration, saturated humidity, and 5% carbon dioxide concentration conditions to induce hypoxia for 36 hours, after that, replace the cell culture medium with no The serum basal culture medium was restored to 21% oxygen concentration, and cultured for 72 hours under saturated humidity and 5% carbon dioxide concentration. After centrifugal filtration, the culture medium was collected, and the calcium alginate composite biofilm was prepared by freeze-drying method.

Embodiment 3

[0029] Example 3 EPC cell culture fluid calcium alginate composite biofilm (Ⅲ)

[0030] The endothelial progenitor cells were subcultured according to the steps of 1) and 2) in Example 1. After culturing the cells to the 5th generation, when the cells grow to 70-80% confluence, place the cells in 1.5% oxygen concentration, saturated humidity, and 5% carbon dioxide concentration conditions to induce hypoxia for 24 hours, after that, replace the cell culture medium with no The serum basal culture solution was restored to 21% oxygen concentration, and cultured for 48 hours under saturated humidity and 5% carbon dioxide concentration. After centrifugal filtration, the culture medium was collected, and the calcium alginate composite biofilm was prepared by freeze-drying method.

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Abstract

The present invention discloses a biofilm capable of rapidly recovering skin wound surface, and a preparation method thereof, wherein the biofilm is prepared from an endothelial progenitor cell serum-free medium liquid and calcium alginate. The specific preparation steps comprise: 1) separating cells from a monocyte layer, inoculating in a culture bottle, carrying out primary culture, completely replacing the culture liquid on the 4th day, and replacing the culture liquid every 2-3 days later; 2) carrying out subculturing after the cells grow to achieve 70-80% fusion; 3) culturing the cells to achieve the 3-5 generation cells, replacing the culture liquid, and carrying out hypoxia induction; 4) carrying out reoxygenation culture on the cells obtained in the step 3) after replacing with a serum-free culture liquid; and 5) carrying out centrifugal filtration, collecting the culture liquid, and preparing a calcium alginate composite biofilm by using a freeze-drying method. According to the present invention, the preparation scheme has characteristics of simpleness, easy performing, low cost, high efficiency, good diabetes wound surface treatment effect, and long-term storage.

Description

technical field [0001] The invention relates to a preparation for promoting the healing of skin wounds, in particular to a biofilm capable of rapidly restoring skin wounds. Background technique [0002] Diabetes has become a chronic disease that seriously threatens human health after tumors and cardiovascular diseases. The biggest threat of diabetes to patients comes from its complications. Among them, diabetic wound has become a serious complication that seriously threatens the survival and quality of life of diabetic patients. It is characterized by refractory, susceptible to infection, and high amputation rate. complex. At present, diabetic wounds are mainly limited to traditional treatment, and the clinical treatment effect is not satisfactory. The effect of some newly developed drugs such as single growth factors is still relatively limited, and there is a lack of simple, economical, safe and efficient new drugs and biological treatments. Therefore, how to effectively...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/44A61K9/70C12N5/071A61P17/02A61P3/10A61K31/734
CPCA61K35/44A61K9/0002A61K9/7007A61K31/734C12N5/0692C12N2500/90A61K2300/00
Inventor 李爽
Owner THE THIRD AFFILIATED HOSPITAL OF GUANGZHOU MEDICAL UNIVERSITY
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