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A kind of gel scaffold transplantation system and its application of composite bone marrow mesenchymal stem cells

A bone marrow mesenchymal and stem cell technology, applied in medical science, prosthesis, coating, etc., to achieve the effect of improving adhesion, promoting neural differentiation, and enhancing adhesion

Active Publication Date: 2016-04-06
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] At present, there is no report on the joint construction of transplantation system of RGD polypeptide, hyaluronic acid and bone marrow mesenchymal stem cells for nerve tissue repair

Method used

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  • A kind of gel scaffold transplantation system and its application of composite bone marrow mesenchymal stem cells
  • A kind of gel scaffold transplantation system and its application of composite bone marrow mesenchymal stem cells
  • A kind of gel scaffold transplantation system and its application of composite bone marrow mesenchymal stem cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Embodiment 1 prepares stent skeleton

[0050] The stent skeleton of this embodiment is prepared through the following steps:

[0051] (1) Add 0.5g sodium hyaluronate (molecular weight 1.5×10 6 Da), stir fully, prepare hyaluronic acid aqueous solution, adjust pH to be 4.5 after observing that dissolves evenly;

[0052] (2) Add ADH to the hyaluronic acid aqueous solution according to the molar ratio of the reactive functional groups (carboxyl group in hyaluronic acid, primary amino group in adipic dihydrazide) HA:ADH=1:4, fully stir to mix evenly, Adjust the pH to 4.5;

[0053] (3) Add EDC to the solution obtained in step (2) according to HA:EDC=1:10, stir to mix quickly and evenly, adjust the pH to 4.5, stir quickly, and immediately inject it into the mold until it forms a gel, adjust pH=7 terminate the reaction;

[0054] (4) The resulting gel was dialyzed with phosphate buffered saline (PBS) with a pH of 7.4, frozen overnight at -80° C. after 48 hours, and freeze-dr...

Embodiment 2

[0056] Same as Example 1, but the ratio of HA to ADH is changed to HA:ADH=1:2, and the ratio of HA to EDC is changed to HA:EDC=1:15.

Embodiment 3

[0058] (1)-(4) are the same as in Example 1, but the ratio of HA to EDC is changed to HA:EDC=1:15.

[0059] (5) Swell the scaffold skeleton in 100mM NaHCO 3 In the solution (pH8.4), after the swelling is sufficient, the acetone gradient is used for dehydration. The volume ratios of acetone / water in each concentration gradient are: 20 / 80, 40 / 60, 60 / 40, 80 / 20, 90 / 10, 95 / 5 and pure acetone;

[0060] (6) Place the dehydrated scaffold skeleton in 100 mg / mL CDI solution for activation for 15 minutes, wash off the acetone with NaHCO3 solution and re-swell the scaffold skeleton;

[0061] (7) Place the stent skeleton after swelling in NaHCO containing 200mM RGD peptide 3 The peptide modification reaction was carried out in the solution (100mM, pH8.4), and the scaffold skeleton was taken out after 48h of reaction, dialyzed for 48h, frozen at -80°C overnight, and freeze-dried to obtain a gel scaffold;

[0062] (8) Swell the gel scaffold in phosphate buffered saline (PBS) with pH = 7.4...

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Abstract

The invention discloses a gel scaffold transplantation system of composite bone marrow mesenchymal stem cells and an application of the gel scaffold transplantation system. The transplantation system comprises a gel scaffold and bone marrow mesenchymal stem cells adhered on the gel scaffold. The preparation method of the gel scaffold is as follows: in the presence of a cross-linking agent, carrying out a cross-linking reaction between HA and ADH, and carrying out freeze drying to obtain a scaffold skeleton, wherein the molar ratio of HA to ADH to the cross-linking agent is 1:(2-4):(7-15) counted by reaction functional groups; dewatering, activating and swelling the scaffold skeleton, modifying the swelled scaffold skeleton by using RGD peptide, and freeze drying to obtain the gel scaffold. According to the invention, the gel scaffold is prepared under a certain raw material ratio, such that the gel scaffold has a high porosity and has a relatively regular three-dimensional reticulate structure; the scaffold skeleton is modified by the RGD peptide to greatly perfect the adhesivity of the stem cells on the gel scaffold, so that the transplantation system has better compatibility with the central nervous tissue.

Description

technical field [0001] The invention belongs to the fields of preparation material preparation and regenerative medicine, and in particular relates to a gel scaffold transplantation system compounded with bone marrow mesenchymal stem cells and an application thereof. Background technique [0002] Central nervous system (CNS) diseases such as chronic degenerative diseases and various injuries such as spinal cord injuries often lead to the loss of neurons, axons and glial cells, resulting in sensory or motor dysfunction or even paralysis. Due to the poor regenerative ability of the CNS after degeneration or injury, it is difficult to generate new neurons to complete self-repair, so stem cell transplantation has become the main trend in the research of nerve tissue repair and regeneration. The first clinical trial using embryonic stem cell transplantation to treat human spinal cord injury was approved by the US FDA in 2009. However, the research and application of embryonic st...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61L27/20A61L27/38A61L27/34A61L27/52A61L27/54
Inventor 高建青李黎明韩旻陈芝兰皇甫铭一刘惠娜
Owner ZHEJIANG UNIV
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