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Active extracellular matrix, composition containing active extracellular matrix, 3D tissue repair scaffold, preparation method and application

A tissue repair and extramatrix technology, applied in tissue regeneration, medical science, prosthesis, etc., can solve problems such as lack of processing capacity, insufficient content of tissue repair materials, and transmission of large-scale diseases with quality variability.

Active Publication Date: 2020-12-29
SHANGHAI EAST HOSPITAL EAST HOSPITAL TONGJI UNIV SCHOOL OF MEDICINE +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This type of ECM has the disadvantage of high quality variability and the risk of disease transmission due to the difficulty in controlling the quality variation of the donor source
Also, the machinability of ECM is a serious issue
Tissue-derived ECMs lack processing capabilities, they cannot be processed into the desired shape, size and porosity without losing their overall structural integrity
[0009] In addition, although the extracellular matrix itself contains some active factors, its content is not enough as a tissue repair material

Method used

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  • Active extracellular matrix, composition containing active extracellular matrix, 3D tissue repair scaffold, preparation method and application
  • Active extracellular matrix, composition containing active extracellular matrix, 3D tissue repair scaffold, preparation method and application
  • Active extracellular matrix, composition containing active extracellular matrix, 3D tissue repair scaffold, preparation method and application

Examples

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Effect test

Embodiment 1

[0066] The preparation method of active extracellular matrix, the method adopts the method of gene modification to introduce gene fragments expressing active factors into cells, make it secrete extracellular matrix and active factors by way of in vitro culture, and then remove cells to obtain active extracellular matrix The extracellular matrix of factors; wherein, the cells are at least one of human cells or other mammalian cells. Preferably, the cells are at least one of human mesenchymal stem cells, human iPS stem cells, and stem cells of other mammals. Using the combined application of the Tet-on expression regulation system and the CRISPR / Cas9 system, the gene fragments of the active factors are introduced into the stem cells during the cell culture process, and the stem cells are expanded in vitro, and the specific overexpression is conditionally overexpressed during the directed differentiation stage of the stem cells. Cell growth factor, promotes the directional differ...

Embodiment 2

[0073] The lentiviral Tet-on vector of bone morphogenic protein BMP2 / BMP4 was constructed in vitro, and the target gene sequence was introduced into the cells through the lentivirus and other gene vectors, and the cells were amplified in large quantities, and then planted on the 3D porous scaffold for culture and amplification. In the stage of inducing cartilage differentiation and bone differentiation, by adding tetracycline (Tet) or doxorubicin (Dox), the overexpression of BMP2 / BMP4 is induced to promote cartilage formation and osteogenesis differentiation. The decellularized ECM will contain a higher content of BMP2 / BMP4, so it can be used in clinical cartilage repair and bone repair materials. Using the Tet-on expression regulation system, the cell expansion can be unaffected, and the specific cell growth factor BMP2 / BMP4 can be conditionally overexpressed in the stage of cell directed differentiation.

[0074] Meanwhile, this example provides a method for decellularizatio...

Embodiment 3

[0076]Human fibroblasts and umbilical vein-derived endothelial cells (EC) were used for co-culture, and the formation of a 3D tubular network structure with a lumen of EC cells up to 400 μm was observed. The tissue-like structures produced by this co-culture technique have the potential to promote vascularization of ECM-coated 3D constructs after implantation. It is well known that vascularization of implants for tissue repair is one of the major challenges in the field of regenerative medicine. In this case, the Tet-on expression regulation system can be used for human fibroblasts, and tetracycline (Tet) or doxorubicin (Dox) can be added at the time of co-culture, so that human fibroblasts can overexpress endothelial cell growth Factor (EGF), which further promotes EC cells to form vascular lumen tissue.

[0077] This active ECM or 3D tissue culture scaffold with active ECM can be mass-produced in advance, then dried by freeze-drying and refrigerated storage, so the product ...

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Abstract

The invention discloses an active extracellular matrix, a composition containing the active extracellular matrix, a 3D tissue repair scaffold, and a preparation method and application. The method comprises the following steps of introducing a gene segment for expressing an active factor into a cell in advance, and carrying out in-vitro cell culture to obtain the extracellular matrix containing theactive factor, so that the reliability of cell origin is ensured while imparting stability and modifiability or processability to the quality of extracellular matrix products. According to the method, stem cell culture and directional differentiation are carried out on a 3D printed porous scaffold to prepare a bionic scaffold capable of promoting damage repair, a material design principle and a stem cell differentiation technology based on stem cell combined directional differentiation are established, and a new field of stem cell and material combination in in-situ tissue engineering repairapplication is developed. According to the method, the bionic scaffold is further optimized, induced adsorption, growth and differentiation of cells after implantation are achieved, and tissue (bone,cartilage, skin, myocardium and blood vessels) regeneration is achieved.

Description

technical field [0001] The invention belongs to the technical field of tissue engineering, and relates to a method for preparing extracellular matrix by in vitro cell culture, and related implants prepared by taking extracellular matrix as a core component, specifically active extracellular matrix, containing active extracellular matrix Composition, 3D tissue repair scaffold, preparation method and application of the invention. Background technique [0002] Organ / tissue damage is one of the most difficult and costly problems in medicine. Due to disease, trauma or congenital problems, there is a large and urgent need for the repair of tissues and organs. Every year, tens of millions of patients suffer from various organ and tissue-related diseases, including skin, bone, cartilage, nerve, heart and kidney injuries, due to trauma, degenerative diseases and burns. Millions of surgeries are performed each year to treat these patients, and the cost of treatment is enormous and c...

Claims

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Application Information

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IPC IPC(8): A61L27/36A61L27/54A61L27/56A61L27/60A61L27/50
CPCA61L27/3633A61L27/54A61L27/56A61L27/60A61L27/507A61L2430/02A61L2430/06A61L2430/22A61L2430/20A61L2430/40A61L2300/412A61L2300/414
Inventor 刘青刘中民乐文俊付艳宾王祎龙孟潇
Owner SHANGHAI EAST HOSPITAL EAST HOSPITAL TONGJI UNIV SCHOOL OF MEDICINE
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