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Injectable hydrogel and preparation method thereof

A technology for injecting water and gel, which is applied in the fields of biomedical materials and tissue engineering, can solve the problems of uncontrollable degradation rate, complicated operation, and destruction of three-dimensional matrix cross-linked network, etc., and achieve the effect of controllable degradation rate

Active Publication Date: 2020-02-21
易小玉
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, during the dissolution process, the three-dimensional matrix cross-linked network of ECM proteins will be destroyed, and there are various active enzyme components that can degrade ECM protein components in the implanted body, such as matrix metalloproteinases and cathepsins, etc. Therefore, injection The pure extracellular matrix implanted in the body will be degraded rapidly in a short period of time, and its degradation rate is uncontrollable, so its function as a bioscaffold in vivo will be limited
[0005] To overcome these limitations, new methods have been developed to fabricate internal vascular structures using solid free-form synthetic materials such as polymers. Polyethylene glycol (PEG) is a well-biosoluble polymeric compound widely used in In the fields of medicine, food and industry, PEG has good hydrophilicity. In the field of medicine, PEGylation of proteins can increase the water solubility of antibodies or recombinant proteins, and at the same time reduce the clearance rate of these protein drugs in the blood, and the synthesis is linear. In addition, multi-arm branched chain structures can also be synthesized, and different functional groups can be modified on the multi-arm branched chain structure to make PEG form various self-crosslinking hydrogels. The gel lacks good biological induction and cannot effectively induce tissue regeneration in the body
[0006] A series of collagen scaffold-based tissue engineering products are currently being developed, that is, collagen-based biomaterials form dispersions of fibers, films, sheets, sponges, and fibrils, and these forms of biomaterials can potentially be used as scaffolds for tissue engineering, Used to repair or enhance body tissues, but in many practical tissue engineering applications, the structure and proteolysis susceptibility of reconstituted protein hydrogels in bioscaffold systems still have many shortcomings compared with native tissues, besides In addition, the method of improving the physical properties of collagen gels based on covalent cross-linking in the prior art is relatively complicated, requiring thiolation of natural proteins first, and UV irradiation or additional initiators and photoinitiators to induce

Method used

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  • Injectable hydrogel and preparation method thereof
  • Injectable hydrogel and preparation method thereof
  • Injectable hydrogel and preparation method thereof

Examples

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Embodiment 1

[0033] A preparation method of injectable hydrogel, comprising the following steps:

[0034] (1) The adipose tissue was repeatedly frozen and thawed 3 times, then soaked in 0.4mol / L NaCl hypertonic liquid for 4 hours, and then soaked in 0.9mol / L NaCl for 4 hours to break the fat cells; The trypsin solution was digested at 35°C for 3 hours, washed with deionized water for 3 hours, treated with isopropanol for 12 hours to remove fat, and then treated with 0.9 wt% sodium lauryl sulfate for 2 hours, and then washed with 0.9% polyethylene glycol octylphenyl ether was treated for 48 hours to perform decellularization to obtain an acellular matrix, which was freeze-dried at minus 80°C and 0.038mbar for 36 hours for use;

[0035] (2) Enzymolyze the lyophilized acellular matrix with 0.5 mg / mL pepsin in an acidic environment for 12 hours, then inactivate the pepsin to obtain a 6 mg / mL extracellular matrix protein solution for use;

[0036] (3) The extracellular matrix protein solution ...

Embodiment 2

[0038] Such as figure 1 Shown, a kind of preparation method of injectable hydrogel comprises the following steps:

[0039] (1) The adipose tissue was repeatedly frozen and thawed 4 times, then soaked in 0.5mol / L NaCl hypertonic liquid for 4 hours, and then soaked in 1mol / L NaCl for 4 hours to break the fat cells; The protease solution was digested at 37°C for 6 hours, washed with deionized water for 4 hours, treated with isopropanol for 18 hours to remove fat, and then treated with 1wt% sodium lauryl sulfate for 18 hours, followed by 1% The polyethylene glycol octyl phenyl ether was treated for 48 hours to perform decellularization to obtain an acellular matrix, which was freeze-dried at minus 80°C and 0.04mbar for 36 hours for use;

[0040] (2) enzymatically hydrolyzing the lyophilized acellular matrix with 3 mg / mL pepsin in an acidic environment for 18 hours, and then inactivating the pepsin to obtain a 20 mg / mL extracellular matrix protein solution for use;

[0041] (3) C...

Embodiment 3

[0043] A preparation method of injectable hydrogel, comprising the following steps:

[0044] (1) The adipose tissue was repeatedly frozen and thawed for 6 times, then soaked in 0.6mol / L NaCl hypertonic liquid for more than 4 hours, and then soaked in 1.1mol / L NaCl for 5 hours to break the fat cells; then use 0.26wt% The trypsin solution was digested at 38°C for 10 hours, washed with deionized water for 4 hours, treated with isopropanol for 24 hours to remove fat, and then treated with 1.1wt% sodium lauryl sulfate for 24 hours. Treat with 1.1% polyethylene glycol octylphenyl ether for 50 hours to perform decellularization treatment to obtain an acellular matrix, and freeze-dry the acellular matrix at minus 80°C and 0.042mbar for 38 hours for use;

[0045] (2) enzymatically hydrolyzing the lyophilized acellular matrix with 5 mg / mL pepsin in an acidic environment for 24 hours, and then inactivating the pepsin to obtain a 40 mg / mL extracellular matrix protein solution for use;

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Abstract

The invention discloses an injectable hydrogel and a preparation method thereof. The preparation method comprises the following steps: sequentially performing cell crushing, primary proteolysis, cleaning, degreasing treatment and decellularization treatment on biological tissue so as to obtain a decellularized matrix, and performing freeze-drying on the decellularized matrix for later use; performing secondary proteolysis on the freeze-dried decellularized matrix so as to obtain a 6-40mg / mL water-soluble extracellular matrix protein solution; and crosslinking the extracellular matrix protein solution with 10-100mg / ml multi-arm modified polyethylene glycol containing a succinimide group, and / or a sulfydryl group, and / or a maleimide group and / or an amino group, so as to obtain the njectablehydrogel. The njectable hydrogel is prepared by using the method. The preparation method disclosed by the invention is simple, and the prepared hydrogel not only has the biological inductivity of collagen, but also has the property of controllable degradation rates of high polymer materials.

Description

technical field [0001] The invention belongs to the technical field of biomedical materials and tissue engineering, and in particular relates to an injectable hydrogel used as an in vivo biological scaffold and a preparation method thereof. Background technique [0002] Tissue engineering is an emerging discipline that combines cell biology and material science to construct tissues or organs in vitro or in vivo. Classical tissue engineering approaches require the use of living cells and basic scaffolds for cell culture to mimic and replace the natural structure of tissue while providing temporary functional support to the cells. [0003] As we all know, there is a complex extracellular matrix (ECM) between most mammalian cells, which is a macromolecule synthesized by animal cells and secreted extracellularly, distributed on the cell surface or between cells. The composition can be divided into three categories: ① glycosaminoglycans (glycosaminoglycans), proteoglycans (prote...

Claims

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Application Information

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IPC IPC(8): A61L27/18A61L27/22A61L27/50A61L27/52A61L27/58C08J3/075C08J3/24
CPCA61L27/227A61L27/18A61L27/50A61L27/58A61L27/52C08J3/075C08J3/246A61L2400/06C08J2389/00C08J2371/02C08L89/00C08L71/02
Inventor 易小玉
Owner 易小玉
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