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A kind of hydrogel with spleen extracellular matrix as raw material and preparation method thereof

A technology of spleen cells and extracellular matrix, which is applied in the fields of medical formula, medical science, prosthesis, etc., can solve the problem of far-flung extracellular matrix, improve the immune microenvironment, have a good application prospect, and promote the formation of lymphatic vessels Effect

Active Publication Date: 2020-01-03
ARMY MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these modified traditional hydrogels are still far from the natural extracellular matrix in terms of ultrastructure and biological function.

Method used

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  • A kind of hydrogel with spleen extracellular matrix as raw material and preparation method thereof
  • A kind of hydrogel with spleen extracellular matrix as raw material and preparation method thereof
  • A kind of hydrogel with spleen extracellular matrix as raw material and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1 Preparation of spleen-specific hydrogel of the present invention

[0022] (1) Decellularization: rewarm the spleen tissue of the same or heterogeneous source (such as pig) stored in the refrigerator slowly for 5-8 hours, remove the fatty tissue such as the hilum of the spleen, and cut it into 1-3mm 3 Soak the small pieces of spleen tissue in saline and rinse, then soak the small pieces of spleen tissue in decellularization solution (0.05%-0.3% SDS+1.5%-4% EGTA, and adjust the pH to neutral), and place at 37°C , shaken at 80r / min for 20-40 hours, replaced the decellularized solution, placed at 25°C, shaken at 60r / min for 30-50 hours, and obtained decellularized spleen extracellular matrix.

[0023] (2) Enzyme digestion: rinse the decellularized spleen extracellular matrix in step (1) with deionized water at 25°C and 80 rpm (change the water every 40 to 60 minutes, 15 to 25 times in total), remove The remaining decellularized liquid was vacuum freeze-dried (the...

Embodiment 2

[0026] Example 2 Detection of the properties of the spleen-specific hydrogel of the present invention

[0027] 1. Detection of decellularized spleen ECM tissue

[0028] 1.1 Tissue Embedding Section

[0029] (1) The extracellular matrix of the spleen treated with decellularization in Example 1 and the untreated spleen tissue were taken, fixed with 4% paraformaldehyde for 48 hours, and washed with running water overnight.

[0030] (2) Dehydration was carried out with graded alcohol, and the ETC was transparent overnight.

[0031] (3) Use Tissue Tek O.C.T. for frozen embedding, and cut into thin slices of about 10 μm.

[0032] 1.2 H&E staining

[0033] (1) Bake the above-mentioned slices at 37°C for a constant length, and bake at 65°C for more than 2 hours before staining.

[0034] (2) ETC dewaxing for 10 minutes, rehydration with gradient alcohol, and washing with running water.

[0035] (3) Hematoxylin staining for 15 minutes, washing with running water for 3 times; color ...

Embodiment 3

[0061] Example 3 The cultivation of macrophages seeded in spleen hydrogel

[0062] 1. Isolation, culture and differentiation of macrophages from peripheral blood

[0063] (1) Take the peripheral blood of the rat, and separate the macrophages of the rat by gradient centrifugation.

[0064] (2) Macrophages were inoculated into medium (10% FBS, 1% penicillin or streptomycin, 2mM L-glutamine and 1mM sodium pyruvate), placed in a humid environment at 37°C, And feed 5% CO2 culture for 7 days, replace the medium every 48 hours.

[0065] (3) After culturing for 7 days, the culture medium was removed by centrifugation, and culture was continued with basal culture medium (10% FBS, 100 ug / ml streptomycin, 100 U / ml penicillin).

[0066] 2. Scanning Electron Microscopy of Spleen-Specific Hydrogels

[0067] (1) The spleen-specific hydrogel prepared in the above-mentioned Example 1 was incubated at 37° C. for 1 h.

[0068] (2) The above isolated and cultured macrophages were centrifuged ...

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Abstract

The invention discloses hydrogel prepared from a spleen extracellular matrix as a raw material and a preparation method thereof. The hydrogel is prepared from the spleens from same or different resources as raw materials by decellularization, digestion and incubation gelling. The spleen-specific hydrogel has the characteristic of self-assembling, effectively forms a stem cell nest, promotes stem cell migration to the diseased region, promotes stem cell survival, promotes angiogenesis and plays a function. The spleen-specific hydrogel can effectively improve the immunological microenvironment of the diseased region, promote the growth of immune cells and formation of lymphatic vessels, effectively repair the diseased region and has a good application prospect in clinical treatment on multiple serious diseases.

Description

technical field [0001] The invention belongs to the technical field of biomaterials, and in particular relates to a hydrogel using spleen extracellular matrix as a raw material and a preparation method thereof. Background technique [0002] Hydrogel is a polymer material with a three-dimensional network structure that uses water as the dispersion medium and is cross-linked through covalent bonds, hydrogen bonds, or van der Waals forces. Because of its unique viscoelasticity, high water content, and environmental responsiveness, it has broad application prospects in biomedical fields such as tissue engineering, drug sustained release, and biosensors. [0003] Traditional hydrogels can be divided into synthetic polymer hydrogels and natural polymer hydrogels. Synthetic hydrogels are mostly homopolymers and copolymers of acrylamide (AAM) and its derivatives, and homopolymers and copolymers of acrylic acid (AA) and its derivatives. Secondly, there are polyvinyl alcohol (PPA), ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61L27/52A61L27/36A61L27/50A61L26/00
CPCA61L26/0057A61L26/0061A61L26/008A61L27/3633A61L27/3641A61L27/3675A61L27/3679A61L27/3687A61L27/50A61L27/52A61L2400/06A61L2430/20A61L2430/22A61L2430/32
Inventor 朱楚洪刘歌
Owner ARMY MEDICAL UNIV
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