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Injectable bone-repairing hydrogel containing exosomes of human umbilical cord mesenchymal stem cells and preparation method of injectable bone-repairing hydrogel

A mesenchymal stem cell and composite hydrogel technology, applied in the field of injectable bone repair hydrogel containing human umbilical cord mesenchymal stem cell exosomes and its preparation, to achieve convenient operation, efficient treatment method, good exosome slowing The effect of release function

Inactive Publication Date: 2021-01-12
温宁
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In the current research on the use of exosomes to repair bone defects, exosomes are mainly injected to play a role, but this method will cause exosomes to be cleared quickly in the body, and bone repair is a long-term process. cycle process

Method used

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  • Injectable bone-repairing hydrogel containing exosomes of human umbilical cord mesenchymal stem cells and preparation method of injectable bone-repairing hydrogel
  • Injectable bone-repairing hydrogel containing exosomes of human umbilical cord mesenchymal stem cells and preparation method of injectable bone-repairing hydrogel
  • Injectable bone-repairing hydrogel containing exosomes of human umbilical cord mesenchymal stem cells and preparation method of injectable bone-repairing hydrogel

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Experimental program
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Embodiment 1

[0036] Example 1 Extraction of exosomes from human umbilical cord mesenchymal stem cells

[0037] (1) Isolation and culture of umbilical cord mesenchymal stem cells

[0038] Take the umbilical cord 15cm near the fetal section, soak it in PBS containing 10% penicillin and streptomycin, take it out in the ultra-clean bench and place it in a petri dish, rinse the umbilical cord repeatedly with PBS, peel off the adventitia and umbilical vessels, and cut it into pieces with tissue scissors to 1mm 3 Tissue pieces of different sizes were transferred to centrifuge tubes, washed by centrifugation with PBS, and centrifuged at 2500rpm for 5min. Resuspend the tissue pieces with a-MEM complete medium, inoculate them in a cell dish, culture in a 37°C, 5% CO2 incubator, add a-MEM complete medium on the second day, and completely change the medium every 3 days thereafter once. When the cells reached 70-80% confluence, they were digested with 0.25% trypsin for 5 minutes, observed under the m...

Embodiment 2

[0041] The preparation method of embodiment 2 hydrogel

[0042] (1) Preparation of nano-hydroxyapatite

[0043]Dissolve 5g of calcium nitrate in 100ml of deionized water, add 0.4g of sodium citrate, stir until completely dissolved, adjust the pH of the solution to 11 with 5mol / L ammonia water, and obtain solution A; dissolve 1.675g of diammonium hydrogen phosphate in 60ml of deionized water Obtain solution B in ionized water; then start the high-gravity rotating packed bed (RPB), adjust the rotation speed to 2500rpm, and transport the two solutions of A and B to the RPB respectively. 10min, the resulting nano-HAP dispersion. Vacuum drying to obtain nHAP powder.

[0044] (2) Preparation of hyaluronic acid derivatives

[0045] Dissolve 1g of hyaluronic acid in 200ml of deionized water, add 12.5g of adipic acid dihydrazide (ADH) after dissolving and stir evenly, then add 2g of 1-(3-dimethylaminopropyl)-3- Ethylcarbodiimide hydrochloride (EDC) catalyzed the reaction, then adju...

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Abstract

The present invention relates to injectable bone-repairing hydrogel containing exosomes of human umbilical cord mesenchymal stem cells and a preparation method of the injectable bone-repairing hydrogel, and belongs to the technical field of biomedical engineering. The hydrogel material contains exosomes of human umbilical cord mesenchymal stem cells, nano-hydroxyapatite, a hyaluronic acid-adipic dihydrazide derivative and an alginic acid derivative with an aldehyde group on a main chain. The preparation method comprises the steps of separation of human umbilical cord mesenchymal cells, extraction of exosomes, preparation of nano-hydroxyapatite, preparation of a hyaluronic acid-adipic dihydrazide derivative, preparation of composite hydrogel and the like. The injectable bone-repairing hydrogel provides a new repair method for a bone defect treatment and has a wide application prospect and an ideal repair effect.

Description

technical field [0001] The invention relates to the technical field of biomedical engineering, in particular to an injectable bone repair hydrogel containing human umbilical cord mesenchymal stem cell exosomes and a preparation method thereof. Background technique [0002] Bone defect repair is a hot topic in the field of orthopedics. Clinically, even for common fractures, about 2%-10% of fractures may progress to nonunion due to poor bone regeneration. Orthopedic clinicians often encounter problems such as bone defects caused by fractures, bone infections, and bone tumor resections. In addition, obesity, diabetes, etc. lead to an increase in musculoskeletal diseases and a decrease in bone regeneration. Elderly patients with osteoporosis have a higher risk of fracture and insufficient bone regeneration capacity. Therefore, how to promote bone regeneration remains a major challenge for clinicians. [0003] Bone tissue engineering technology based on mesenchymal stem cells ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/52A61L27/58A61L27/20A61L27/12A61L27/22C08J3/075C08L5/04C08L5/08C08K3/32
CPCA61L27/12A61L27/20A61L27/227A61L27/52A61L27/58A61L2300/252A61L2300/412A61L2430/02C08J3/075C08J2305/04C08J2405/08C08K2003/325C08K2201/011C08L5/04C08L5/08
Inventor 杨烁张建军朱彪赵立升付之光温宁
Owner 温宁
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