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Fibrin hydrogel stent loaded with human umbilical cord mesenchymal stem cells and application of fibrin hydrogel stent

A technology of stem cells and fibrin, applied in the field of fibrin hydrogel scaffolds, can solve the problems of high cost, long time, cumbersome preparation process, etc., achieve the effect of convenient material collection, promotion of re-epithelialization, and meeting treatment needs

Pending Publication Date: 2021-01-26
THE FIRST AFFILIATED HOSPITAL OF SOOCHOW UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the preparation process of the existing technology is cumbersome, time-consuming, and costly, and cannot solve the problem that stem cells are difficult to survive in the wound

Method used

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  • Fibrin hydrogel stent loaded with human umbilical cord mesenchymal stem cells and application of fibrin hydrogel stent
  • Fibrin hydrogel stent loaded with human umbilical cord mesenchymal stem cells and application of fibrin hydrogel stent
  • Fibrin hydrogel stent loaded with human umbilical cord mesenchymal stem cells and application of fibrin hydrogel stent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1: Primary isolation and culture of human umbilical cord mesenchymal stem cells

[0028] After the human umbilical cord is obtained from the operating table, it is immersed in PBS preservation solution containing 1% double antibody (penicillin) under aseptic conditions, the 50ml centrifuge tube is sealed and sterile, and it is transported to the laboratory in a 4°C ice box. Take it out in the ultra-clean workbench, wash it twice with 0.9% normal saline, remove the blood stasis at both ends, fully wash the periphery of the umbilical cord and the inner cavity of the umbilical vein with PBS buffer containing 1% double antibody, and remove the arteriovenous For blood vessels, take the umbilical cord matrix and cut into 4-5mm 3 Tissue pieces of different sizes were inoculated into T25 culture flasks containing 1ml DMEM / F12 (1:1) medium (containing 10% FBS), 5 pieces in each bottle, and a total of 10 bottles were placed in 37°C, 5% CO 2 , Culture in a saturated humi...

Embodiment 2

[0029] Example 2: Preparation of fibrin hydrogel scaffold loaded with human umbilical cord mesenchymal stem cells

[0030] A preparation method of a fibrin hydrogel scaffold loaded with human umbilical cord mesenchymal stem cells, specifically comprising the following steps:

[0031] (1) Human umbilical cord mesenchymal stem cells are subcultured after primary isolation and culture, and the subcultured P1-P6 generation human umbilical cord mesenchymal stem cells are cultured in culture flasks in a carbon dioxide incubator to make their density reach 70%-80%.

[0032] (2) Dissolving fibrinogen in sterile water for injection to prepare a fibrinogen solution (10 mg / ml).

[0033] (3) Dissolve thrombin in calcium ion solution (including 300mmol / L NaCl, 40mmol / L CaCl2) to prepare thrombin solution (25U / ml).

[0034] (4) Human umbilical cord mesenchymal stem cells were digested with 0.25% Trypsin-EDTA, and the resulting cells were resuspended with fibrinogen solution.

[0035] (5)...

Embodiment 3

[0036] Embodiment 3: Mouse full-thickness skin wound experiment

[0037] SPF grade BALB / C mice were used to construct a full-thickness skin wound model, and a fibrin hydrogel scaffold loaded with human umbilical cord mesenchymal stem cells was locally injected into the wound to evaluate its effect on skin wound healing.

[0038] (1) SPF grade BALB / C nude mice (8 weeks old, body weight 18-20g, female) were randomly divided into human umbilical cord mesenchymal stem cells + fibrin hydrogel group, human umbilical cord mesenchymal stem cell group, fibrin hydrogel group, Hydrogel group, saline group.

[0039] (2) Use an isoflurane gas anesthesia machine to induce anesthesia on mice with high concentration and high oxygen flow. After the anesthesia is satisfactory, fix them in the prone position on the operating table, maintain anesthesia at low concentration and low oxygen flow, and disinfect the back skin with Aner Iodine disinfectant three times.

[0040] (3) After laying a ste...

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Abstract

The invention discloses a fibrin hydrogel stent loaded with human umbilical cord mesenchymal stem cells and application of the fibrin hydrogel stent, and belongs to the technical field of hydrogel. The human umbilical cord mesenchymal stem cells are cultured until the density reaches 70 percent to 80 percent; treatment is performed by using a cell digestion solution, and the human umbilical cord mesenchymal stem cells are resuspended by using a fibrin raw solution to obtain a human umbilical cord mesenchymal stem cell-fibrin raw solution mixed solution; and a thrombin solution is added into the mixed solution to obtain the fibrin hydrogel stent loaded with the human umbilical cord mesenchymal stem cells. Through combined application of the fibrin hydrogel stent and the human umbilical cordmesenchymal stem cells, the healing speed of a wound is obviously accelerated; the re-epithelialization is promoted; the inflammatory response is inhibited; the angiogenesis is promoted; the scar formation is reduced; the wound is healed to be close to normal skin tissues to a greater extent; and the treatment requirement is met.

Description

technical field [0001] The invention relates to a fibrin hydrogel scaffold loaded with human umbilical cord mesenchymal stem cells and an application thereof, belonging to the field of hydrogel technology. Background technique [0002] Impaired wound healing has become a global public health challenge, imposing enormous psychological, financial, and emotional burdens on patients and physicians. According to the statistics of the World Health Organization, about 180,000 people die from severe burns every year in the world. The high mortality rate in burn patients is due to increased metabolic demands due to skin loss, severe fluid loss and increased risk of infection. Others, such as diabetic patients, whose wounds heal slowly or even fail to heal, also have a high risk of wound infection and other complications. Skin grafting, as a first-line treatment for wound healing, has deficiencies such as limited natural skin and inconsistent skin color. [0003] In recent years, s...

Claims

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Application Information

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IPC IPC(8): A61L27/22A61L27/38A61L27/50A61L27/52A61L27/58A61L27/60
CPCA61L27/52A61L27/58A61L27/50A61L27/225A61L27/3834A61L27/60A61L27/3895
Inventor 陈友国胡吕忠周金华张琳沈芳荣王娟
Owner THE FIRST AFFILIATED HOSPITAL OF SOOCHOW UNIV
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