Tissue engineering patch and preparation method thereof

A tissue engineering and patch technology, used in tissue regeneration, medical science, prosthesis, etc., can solve problems such as uneven distribution, cell damage, and sticking to the surface of scaffold materials, achieve high target cell load, increase tissue Compatibility, the effect of facilitating immigration

Active Publication Date: 2021-03-30
JINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The advantage of this method is that the use of gel to encapsulate the seed cells provides a three-dimensional void structure for cell growth and proliferation while providing protection for the cells, but the disadvantage is also obvious that the gel can only be attached to the surface of the scaffold material, and cannot be evenly distributed. throughout the scaffold material, and the gel forms a resistance for cell migration into the interior of the scaffold
The dynamic culture system method is to immerse the scaffold material in a high-density cell suspension, and use rotation, centrifugation, and shaking culture operations to inoculate the cells on the scaffold material. Compared with the immersion method and precipitation method, the dynamic culture system can make the cells more uniform. And more adsorbed on the scaffold material and more conducive to the migration of cells to the interior of the scaffold material, but at the same time as the physical factors imposed by the dynamic culture system, rotation, centrifugation and shaking will also cause loosely adhered cells to fall off the scaffold, Or cause damage to the cells, affecting the inoculation rate of the seed cells and the activity of the seed cells
Although the above cell construction techniques can achieve the purpose of compounding seed cells and scaffold materials, the cell load of the scaffold material after inoculation is low, the cells are unevenly distributed in the scaffold material, and how to ensure the quality of seed cells during the inoculation process is still a problem. issues that need resolving
[0004] Although tissue engineering patch technology has a wide range of application scenarios and bright prospects, the insufficiency of existing construction technologies limits the application and therapeutic effect of tissue engineering patch

Method used

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  • Tissue engineering patch and preparation method thereof
  • Tissue engineering patch and preparation method thereof
  • Tissue engineering patch and preparation method thereof

Examples

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preparation example Construction

[0041] This embodiment provides a method for preparing a tissue engineering patch, comprising the following steps:

[0042] (1) Inoculate the target cells on one side of the decellularized scaffold, place pyroptotic products on the other side of the decellularized scaffold, and separate the pyroptotic products from the surface of the other side of the decellularized scaffold; The decellularized scaffold inoculated with the target cells and the pyroptosis product are cultured together in the culture medium;

[0043] (2) When the target cells migrate from one side of the decellularized scaffold to the other side, remove the non-migrated target cells on the surface of the decellularized scaffold to obtain a tissue engineering patch;

[0044] The pyroptotic product is at least one of pyroptotic secretion, pyroptotic extract and pyroptotic cells produced after pyroptosis.

[0045] The pyroptosis of the present invention refers to programmed cell inflammatory necrosis, which is cha...

Embodiment 1

[0060] Example 1 Construction of tissue engineered adipose tissue in vitro

[0061] (1) Pre-cultivate non-target cells to obtain an inflammatory environment

[0062] Bone marrow mesenchymal stem cells were used as non-target cells at 1*106cell / cm 2 The density was inoculated on a large dish with a diameter of 10 cm, and cultured in a carbon dioxide incubator for 24 hours. The medium was a complete medium: low-sugar DMEM+10% fetal bovine serum+1% glutamine+1% penicillin-streptomycin ( 100X). After 24 hours, replace the medium with a complete medium containing 2 mol / ml lipopolysaccharide, and culture for 36 hours. After bone marrow mesenchymal stem cells form obvious pyroptotic bodies (such as figure 2 As shown, the spherical object framed by the red box is the pyroptotic body, and the black arrow points to the pyroptotic cell), indicating that the culture of bone marrow mesenchymal stem cells induced pyroptosis is completed. The medium was removed, and the bottom of the dis...

Embodiment 2

[0077] Example 2: Construction of tissue engineered liver tissue in vitro

[0078] (1) Perform pyroptotic pre-stimulation on non-target cells to obtain an inflammatory environment.

[0079] Isolate cardiosphere stem cells (CDCs), use cardiosphere stem cells as non-target cells, and use 5*106cell / cm 2 The density was inoculated on OPC (hydroxypropyl cellulose octanoate)-coated large dishes and cultured for 3 days. Three days later, the secretion of pyroptotic inflammatory factors increased, pyroptotic bodies were formed, and the pyroptotic marker caspase-1 was stained (such as image 3 ), indicating that the pyroptotic induction culture of cardiomyocyte stem cells was completed, and a large dish containing pyroptotic cardiosphere stem cells was obtained.

[0080] (2) Obtain a single-layer or multi-layer cell sheet of the target cells, and inoculate the obtained cell sheet on one side of the decellularized scaffold.

[0081] After suspending bone marrow mesenchymal stem cells...

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Abstract

The invention relates to a tissue engineering patch and a preparation method thereof, and the preparation method comprises the following steps: (1) inoculating target cells on the surface of one sideof an acellular scaffold, placing a pyroptosis product on the other side of the acellular scaffold, and separating the pyroptosis product from the surface of the other side of the acellular scaffold;culturing the decellularized scaffold inoculated with the target cells and a pyroptosis product in a culture solution together; (2) when the target cells migrate from one side of the decellularized scaffold to the other side of the decellularized scaffold, removing the target cells which are not migrated on the surface of one side of the decellularized scaffold to obtain a tissue engineering patch; wherein the cell pyroptosis product is at least one of cell pyroptosis secreta, a cell pyroptosis extract and pyroptosis cells generated after cell pyroptosis. The tissue engineering patch preparedby the method has high cell loading capacity, target cells are uniformly distributed in the scaffold, and the tissue engineering patch has good biological activity.

Description

technical field [0001] The invention relates to the field of tissue engineering, in particular to a tissue engineering patch and a preparation method thereof. Background technique [0002] Stem cell therapy has achieved good therapeutic effects both experimentally and clinically, showing bright application prospects. At present, the methods of using stem cells or delivering stem cells in the field of stem cell therapy are mainly divided into three categories: 1. Direct injection of stem cells; 2. Stem cell sheet technology; 3. Tissue engineering patch technology. These three types of treatment methods have their own advantages and disadvantages. The use of stem cell injection is simple and easy to repeat, causing little damage to patients, but the effective use rate of stem cells is low, especially when intravenous injection is used as the delivery method of stem cells, most of the stem cells cannot be in the damaged tissue. Play a therapeutic role, and due to the inflammat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/36A61L27/38A61L27/52A61L27/22A61L27/24
CPCA61L27/22A61L27/225A61L27/24A61L27/3604A61L27/3834A61L27/52A61L2430/20A61L2430/28A61L2430/40
Inventor 武征张中夏梁锦超林熙张建华
Owner JINAN UNIVERSITY
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