Tissue engineering scaffold of composite exosome nidogen-1 and its preparation method

A technology of composite tissue and exosomes, applied in the field of medical biomaterials, can solve problems such as ischemia, hypoxia, and slow metabolism, and achieve the effect of promoting angiogenesis, accelerating the process of angiogenesis, and accelerating the repair of bone defects

Active Publication Date: 2021-10-29
FOURTH MILITARY MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In order to solve the problems of ischemia, hypoxia and slow metabolism in tissue engineering grafts, the present invention provides a composite exosome Nidogen-1 based on the first confirmation that the structural protein Nidogen-1 exists in exosomes and has the function of regulating proteins. 1 tissue engineering scaffold and preparation method thereof

Method used

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  • Tissue engineering scaffold of composite exosome nidogen-1 and its preparation method
  • Tissue engineering scaffold of composite exosome nidogen-1 and its preparation method
  • Tissue engineering scaffold of composite exosome nidogen-1 and its preparation method

Examples

Experimental program
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Effect test

Embodiment 1

[0042] 1. Exosome collection

[0043] 1.1 Collection of bone marrow mesenchymal stem cell supernatant or serum

[0044] For bone marrow mesenchymal stem cells (BMSC): select 10-day-old rat pups (purchased from the Animal Experiment Center of Fourth Military Medical University in January 2017), kill them by neck dislocation, take out the femur and tibia, and cut off the metaphysis. Expose the bone marrow cavity, wash the bone marrow with culture medium (a-MEM + 10% fetal bovine serum + 1% penicillin / streptomycin double antibody), collect bone marrow mononuclear cells, separate and purify to P3 generation by adherence method, flow cytometry The expression of the surface markers was detected by surgery, and the cells (CD34-CD45-CD11b / c-CD90+) had a purity greater than 93% and could be used for the next test. The separated and purified cells were inoculated into culture flasks, and the serum-free medium was replaced when the cells were fused to 80%, and the cell supernatant was c...

Embodiment 2

[0079] According to the preparation steps of above example 1 ( figure 1 ), the present invention has actually carried out the experiments of multiple examples for different tissue engineering scaffold material components, and the proportioning ratio of the components in the preparation steps of each example is different, see Table 1 for details.

[0080] For the different examples in Table 1, sodium alginate is an organic compound derived from algae, which is low in price, has good biocompatibility, and is easy to cross-link. Adjusting the ratio of sodium alginate can obtain different mechanical properties (strength , toughness) scaffold material, the optimal ratio can be selected according to the mechanical properties of the repaired target organ. Hyaluronic acid is an acidic mucopolysaccharide with good biocompatibility. Different proportions of hyaluronic acid can be used to prepare repair scaffolds for different organs. For example, to prepare articular cartilage repair sc...

Embodiment 3

[0085] The difference from Example 1 is that the vascular endothelial cells were pre-treated with exosome Nidogen-1 in vitro, and the treated vascular endothelial cells were mixed with the liquid composite hydrogel to construct the tissue for the preparation of bionic bone scaffolds Engineering bracket. The results showed that vascular endothelial cells treated with exosomal Nidogen-1 can also promote cardiovascularization in the bionic bone scaffold, but the disadvantage is that the implantation of the bionic bone scaffold may lead to an immune response.

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Abstract

The invention discloses a tissue engineering scaffold of a composite exosome Nidogen-1 and a preparation method thereof. The supernatant of bone marrow mesenchymal stem cells after expression processing is collected, and the exosome Nidogen-1 is separated by ultracentrifugation, and its content is adjusted. The concentration used is 10‑100 μg / mL; sodium alginate, hyaluronic acid, and PEG‑600 are dissolved in deionized water to make a mixed prepolymer; exosome Nidogen‑1 is added to prepare the composite exosome Nidogen‑ 1 tissue engineering scaffold. Implanting the tissue engineering scaffold into the wound site or the biocompatible tissue engineering material center can promote the regeneration of blood vessels in the defect site and improve the metabolic microenvironment of the material center, which has important clinical application value.

Description

technical field [0001] The invention belongs to the field of medical biomaterials, and in particular relates to a tissue engineering scaffold of composite exosome Nidogen-1 and a preparation method thereof. Background technique [0002] Large-scale tissue and organ defects caused by trauma and disease have the characteristics of high incidence and high disability rate. Continuous breakthroughs in regenerative medicine and tissue engineering technology provide strong support for the use of exogenous materials to transplant and repair defective tissues. However, early vascularization is still the key factor restricting its repair effect. Tissue engineering scaffold products are generally constructed in vitro and applied in vivo. Studies have confirmed that tissue fluid can only penetrate to 300 μm below the surface of the scaffold material. The cells inside the scaffold cannot obtain sufficient oxygen and nutrient supply, and their proliferation, differentiation and secretion...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61L27/10A61L27/18A61L27/20A61L27/22A61L27/52A61L27/54A61L27/56A61L27/58A61L27/60A61K38/17A61P17/02A61P19/08
CPCA61K38/1709A61L27/10A61L27/18A61L27/20A61L27/227A61L27/52A61L27/54A61L27/56A61L27/58A61L27/60A61L2300/252A61L2300/412A61L2400/06A61L2430/02A61L2430/34A61P17/02A61P19/08C08L5/08C08L5/04C08L71/02C08L89/00
Inventor 程朋真刘斌杨柳曹天庆宁芬茹
Owner FOURTH MILITARY MEDICAL UNIVERSITY
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