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Freeze-dried powder used for promoting healing of diabetic wound and preparing method thereof

A technology of wound healing and freeze-dried powder, which is applied in the direction of freeze-dried delivery, metabolic diseases, skin diseases, etc., can solve the problems of slow effect, high cost and unfavorable promotion and application, and achieve reduction of deterioration, low cost, and promotion of angiogenesis and neurogenesis. The effect of regeneration

Inactive Publication Date: 2017-03-22
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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  • Freeze-dried powder used for promoting healing of diabetic wound and preparing method thereof
  • Freeze-dried powder used for promoting healing of diabetic wound and preparing method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1 EPC cell culture fluid freeze-dried powder (I)

[0023] 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 temperature of 37°C.

...

Embodiment 2

[0028] Example 2 EPC cell culture medium freeze-dried powder (Ⅱ)

[0029] 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. The medium was centrifuged, concentrated and lyophilized.

Embodiment 3

[0030] Example 3 EPC cell culture medium freeze-dried powder (Ⅲ)

[0031] 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. The medium was centrifuged, concentrated and lyophilized.

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Abstract

The invention discloses freeze-dried powder used for promoting healing of a diabetic wound and a preparing method thereof. The freeze-dried powder is made by freeze-drying a serum-free nutrient solution in an endothelial progenitor cell. The preparing method specifically comprises the steps of separating cells with mononuclear cell layers, inoculating the cells in a culture flask to be subjected to preliminary cultivation, changing a culture solution in a whole-content mode in the fourth day, and afterwards changing the culture solution once every 2-3 days; conducting continuous cell culture after the cells grows until 70-80% of the cells fuse; culturing the cells to the third to fifth generation, and changing the culture solution to be subjected to hypoxia inducing; conducting reoxygenation cultivation on the inducing cultured cells after changing the culture solution into the serum-free culture solution; conducting centrifugal filtration on the culture solution, collecting the culture solution, and conducing concentration and freeze-drying on the culture solution. The preparing method is simple, convenient and easy to operate, low in cost, high in efficiency and good in wound treating effect of diabetes, and the freeze-dried powder can be stored for a long time.

Description

technical field [0001] The invention relates to a preparation for promoting skin wound healing, in particular to a freeze-dried powder preparation for promoting diabetic wound healing. 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...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/51A61K9/19A61P3/10A61P17/02
CPCA61K35/51A61K9/19
Inventor 李爽
Owner THE THIRD AFFILIATED HOSPITAL OF GUANGZHOU MEDICAL UNIVERSITY
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