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Injectable photo-crosslinking acellular matrix composite hydrogel as well as preparation method and application thereof

A technology of acellular matrix and composite hydrogel, applied in the field of biomaterials, can solve the problems of poor mechanical properties of hydrogel, slow gelation, and difficulty in supporting the shape of surrounding tissues, so as to promote rapid repair and regeneration, and improve mechanical properties , the effect of good operability

Pending Publication Date: 2022-07-08
SUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

After dissolving the separated acellular matrix, the protein components in the solution can be spontaneously recombined into a homogenized hydrogel by adjusting the temperature and pH, but this cross-linking method is slow to form a gel, and the prepared hydrogel Poor mechanical properties, easy to degrade, and difficult to support the shape of surrounding tissues after implantation in the human body, which seriously limits its application in the field of biomedical materials
[0003] In order to improve the mechanical properties of the acellular matrix composite hydrogel, CN105879119A discloses a composite acellular matrix hydrogel and a preparation method thereof, in which a biocompatible three-dimensional fiber network material is used as a skeleton, and is soaked, coated or sprayed The composite hydrogel was prepared by compounding with the acellular matrix precursor solution by the above method. Under the action of the skeleton, the mechanical properties of the composite hydrogel were improved, but there was a distribution of acellular matrix components in the composite hydrogel prepared by the above method. Problems such as unevenness and layered structure
In addition, CN111184914A discloses a functionalized decellularized matrix biomaterial, in which the decellularized animal tissue is sequentially immersed in methacrylic anhydride solution and 3-propyl methacrylate solution for functional modification , to endow the acellular matrix biomaterial with specific functionality, and to form an interpenetrating network structure through free radical polymerization and cross-linking with the acellular matrix biomaterial, which improves the mechanical properties of the acellular matrix material, but this method relies on animal tissue prototypes , and repeated immersion in organic solutions will reduce the biological activity of the acellular matrix, which limits the application of this type of acellular matrix biomaterials

Method used

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  • Injectable photo-crosslinking acellular matrix composite hydrogel as well as preparation method and application thereof
  • Injectable photo-crosslinking acellular matrix composite hydrogel as well as preparation method and application thereof
  • Injectable photo-crosslinking acellular matrix composite hydrogel as well as preparation method and application thereof

Examples

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Embodiment 1

[0045] The present embodiment relates to the preparation of acellular matrix powder, and the specific operations are as follows:

[0046](1) The annulus fibrosus was isolated and extracted from the intervertebral disc of the fresh porcine spine, the nucleus pulposus tissue inside the annulus fibrosus and the fascial adipose tissue outside the annulus fibrosus were removed, and rinsed three times with phosphate buffered saline (PBS). Cut the rinsed annulus fibrosus tissue into about 1mm in size 3 Then, the minced tissue pieces were placed in 0.25% trypsin solution at 37°C under constant stirring for 24 hours, and the solution was changed every 4 hours.

[0047] (2) The tissue block after the above treatment was rinsed 3 times with PBS for 3 minutes each time. Afterwards, the tissue samples were placed in a nuclease solution (50 U / ml DNase and 1 U / ml RNase in PBS) at 37°C with constant stirring for 12 hours.

[0048] (3) The sample was then placed in 1% Triton X-100 solution a...

Embodiment 2

[0053] The present embodiment relates to the preparation of an injectable, high-strength, photocrosslinked acellular matrix composite hydrogel, and the specific operations are as follows:

[0054] Preparation of pregelling solution: 30 mg of pepsin was weighed and dissolved in 30 mL of 0.01 M hydrochloric acid solution, and filtered with a 0.22 μm sterile filter. 300 mg of the acellular matrix powder prepared in Example 1 was added to the pepsin-acetic acid solution, stirred continuously for 48 hours at room temperature, and centrifuged at 8000 rpm for 10 minutes to remove undigested matrix particles to obtain acellular matrix preform. glue solution.

[0055] Preparation of acellular matrix composite hydrogel: The above pregelling solution was placed in a 4°C refrigerator for pre-cooling treatment, and after 5 hours, it was taken out and placed on ice for continuous stirring. Sodium oxide solution, adjust pH to 7.4. Dissolve PEGDA and photoinitiator LAP in pre-cooled PBS, st...

Embodiment 3

[0057] The present embodiment relates to the preparation of an injectable, photocrosslinked acellular matrix composite hydrogel, and the specific operations are as follows:

[0058] Preparation of pregelling solution: Weigh 40 mg of pepsin, dissolve it in 40 mL of 0.01 M hydrochloric acid solution, and filter with a 0.22 μm sterile filter. 400 mg of the acellular matrix powder prepared in Example 1 was added to the pepsin acetic acid solution, stirred continuously for 72 hours at room temperature, and centrifuged at 6000 rpm for 15 minutes to remove undigested matrix particles to obtain acellular matrix preform. glue solution.

[0059] Preparation of acellular matrix composite hydrogel: The above pregelling solution was placed in a 4°C refrigerator for pre-cooling treatment, and after 6 hours, it was taken out and placed on ice for continuous stirring. Sodium oxide solution, adjust pH to 7.3. Dissolve PEGDA and photoinitiator LAP in pre-cooled PBS, stir and mix with the abov...

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Abstract

The invention discloses injectable and photo-crosslinked acellular matrix composite hydrogel as well as a preparation method and application thereof. The preparation method comprises the following steps: (1) mixing acellular matrix powder with an acidic pepsin solution, stirring, and removing undigested matrix particles to obtain an acellular matrix pre-gelatinizing solution; (2) carrying out precooling treatment on the decellularized matrix pregelatinizing solution, adding an alkaline solution, and adjusting the pH value to 7.0-7.4; and (3) dissolving polyethylene glycol diacrylate and a photoinitiator in a pre-cooled buffer solution, uniformly stirring with the pre-gelatinizing solution in the step (2) at 0-8 DEG C, and carrying out photo-crosslinking curing to obtain the acellular matrix composite hydrogel. The preparation method is simple and high in operability, and the prepared composite hydrogel is high in mechanical strength, long in degradation time and good in biocompatibility; the hydrogel can be used for loading growth factors, and illumination crosslinking is performed after the hydrogel is injected into a living body, so that sustainable release of the growth factors in the living body is realized, and tissue repair and regeneration are promoted.

Description

technical field [0001] The invention relates to the technical field of biological materials, in particular to an injectable, photocrosslinked acellular matrix composite hydrogel and a preparation method and application thereof. Background technique [0002] Extracellular matrix (ECM) is a complex network composed of macromolecular substances synthesized and secreted by various tissue cells such as fibroblasts, mesenchymal cells, and epithelial cells, which are distributed and aggregated on the cell surface and between cells. The ECM forms the skeleton of tissues and organs, supports and connects tissue structures, regulates tissue development, and forms the microenvironment required for cell growth. The ECM plays an important role in tissue development, homeostasis, and disease development. The acellular matrix obtained by removing cells from tissues by physical, chemical and biological methods can effectively retain the unique components of the natural ECM, such as collage...

Claims

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Application Information

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IPC IPC(8): A61L27/36A61L27/18A61L27/50A61L27/52A61L27/54A61L27/56
CPCA61L27/3633A61L27/3687A61L27/3691A61L27/18A61L27/50A61L27/52A61L27/54A61L27/56A61L2400/06A61L2430/40A61L2300/414A61L2300/602A61L2300/412C08L71/02
Inventor 魏强韩凤选蔡燕李斌杨惠林
Owner SUZHOU UNIV
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