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Fabrication method of porous micro-scaffold and its application in tissue regeneration and repair

A technology of tissue regeneration and production method, applied in the field of regenerative medicine, can solve the problems of undeveloped stem cell survival, expansion, directed differentiation, injectable three-dimensional porous micro-scaffold, etc., to promote stem cell proliferation, improve adherence performance, small physical effect of size

Active Publication Date: 2019-09-27
PEKING UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] However, injectable three-dimensional porous microscaffolds that can effectively promote stem cell survival, expansion, and directed differentiation have not yet been developed.

Method used

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  • Fabrication method of porous micro-scaffold and its application in tissue regeneration and repair
  • Fabrication method of porous micro-scaffold and its application in tissue regeneration and repair
  • Fabrication method of porous micro-scaffold and its application in tissue regeneration and repair

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0106] Example 1 Preparation of Sodium Alginate Three-Dimensional Porous Microscaffold

[0107] 1.1. Preparation of sodium alginate three-dimensional porous scaffold (AS)

[0108] Sodium alginate (alginate) used in this study was RGD polypeptide (GGGGRGDASSP, purchased from Shanghai Qiangyao).

[0109] Modification method of RGD-alginate (RA):

[0110] Add EDC / NHS to sodium alginate solution, stir at room temperature for 4 hours, add RGD polypeptide and stir well, and then stand at 4°C overnight; then transfer the solution to a dialysis bag (3.5kDa) for dialysis for 3 days, then freeze-dry for 24 hours to obtain RA ; Sealed and stored at -20°C.

[0111] Dissolve 2.0g RA in 100mL deionized water, transfer it to a 24-well plate after fully dissolved, 0.5ML / well, then place the plate in a 4°C refrigerator for 2h, and a -20°C refrigerator for 8h, then freeze-dry for 24h . After freeze-drying was completed, calcium chloride (2%, w / v) solution was added into the well plate, 1 ...

Embodiment 2

[0115] Example 2 Characterization of Sodium Alginate Three-dimensional Porous Microscaffold

[0116] Scanning electron microscopy (SEM; S-4800; Hitachi, Japan) was used to characterize the microstructure (pore size, porous morphology, etc.) To measure the elastic modulus of each scaffold.

[0117] Soak the freeze-dried micro-scaffold in PBS, after it fully absorbs water and swells, filter off the PBS, add a little low-concentration sodium alginate solution to the micro-scaffold and mix well, then transfer to a syringe (1mL size) for freeze-drying . For in vivo experiments, just add the cell suspension to the micro-scaffold and mix well before injection. Injectability was verified with dense tissue.

[0118] Experimental results:

[0119] like figure 1 As shown in A, SEM results show that AMS has a three-dimensional porous structure with high connectivity, and its pore structure is not damaged; as figure 1 As shown in B-1C, the size of the scaffold is about 1 mm, and th...

Embodiment 3

[0122] Embodiment 3 injectable AMS preparation

[0123] The prepared AMS was soaked in PBS at 4°C for 48h; then the PBS was filtered off, the AMS was transferred to a 1ml syringe, the injection plunger was plugged, and sealed and stored at 4°C after freeze-drying.

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Abstract

The invention discloses application of a porous micro stent to regeneration and repair of tissues. The porous micro stent is a sodium alginate three-dimensional porous micro stent; sodium alginate ismodified sodium alginate; the sodium alginate three-dimensional porous micro stent is prepared from liquid nitrogen through a quick freezing and high-speed crushing method; a specific using method ofapplication of the sodium alginate three-dimensional porous micro stent to regeneration and repair of tissues comprises the following steps: adding PBS accounting for 50-80% of the total volume of themicro stent into the prepared micro stent to obtain an injectable micro stent, and injecting the injectable micro stent to injured tissue parts. The porous micro stent is capable of accelerating regeneration of the tissues and efficiently accelerating amplification and directional differentiation of stem cells, is injectable, and is high in clinical practicability. The invention also discloses application of the porous micro stent to acceleration of proliferation of the stem cells, acceleration of differentiation of the stem cells and acceleration of self-regeneration and repair of the tissues.

Description

technical field [0001] The invention relates to the application of a scaffold in tissue regeneration and repair, and belongs to the field of regenerative medicine, in particular to the application of a porous micro-stent in tissue regeneration and repair. Background technique [0002] By simulating the physical and chemical properties of the natural microenvironment of the human body, biological scaffolds can effectively promote the self-renewal, proliferation, and directed differentiation of stem cells, and have been widely used in the field of regenerative medicine in recent years. [0003] Generally, stem cell-based bioscaffolds are mainly divided into porous scaffolds and hydrogel scaffolds according to their state. Among them, the porous scaffold has a large number of interconnected three-dimensional porous structures, which are more conducive to the migration of cells in the scaffold and the supply of nutrients compared to hydrogels. It is generally believed that a th...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61L27/20A61L27/56A61L27/50
Inventor 魏世成罗祖源潘冀佳陈庆林
Owner PEKING UNIV
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