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Agarose-gelatin graft temperature-sensitive hydrogel, and preparation method and application thereof

A temperature-sensitive hydrogel and agarose technology, applied in the medical field, can solve problems such as cell and DNA damage, and achieve the effects of improving biological activity and degradation performance, simple preparation method and low immunogenicity

Inactive Publication Date: 2019-09-10
JINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, some studies have also stated that the key problem of using UV photocuring to cross-link cell-loaded hydrogels is the damage to cells and DNA by UV or UV-induced free radicals (Mellati A, Fan C M, Tamayol A, et al. Microengineered 3D cell- ladenthermore responsive hydrogels for mimicking cell morphology and orientation incartilage tissue engineering.[J].Biotechnology&Bioengineering,2017,114(1):217-231.)

Method used

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  • Agarose-gelatin graft temperature-sensitive hydrogel, and preparation method and application thereof
  • Agarose-gelatin graft temperature-sensitive hydrogel, and preparation method and application thereof
  • Agarose-gelatin graft temperature-sensitive hydrogel, and preparation method and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0049] (1) Preparation of gelatin solution: 1.2 g of gelatin with a molecular weight of 60 kDa was added to 30 mL of dimethyl sulfoxide (DMSO) from which water had been removed, stirred and dissolved at 60°C in an oil bath to obtain a gelatin solution, which was cooled and set aside.

[0050] (2) Preparation of agarose solution: 0.6 g of agarose with a molecular weight of 50 kDa was added to 30 mL of DMSO from which water had been removed, stirred and dissolved in an oil bath at 80° C. to obtain an agarose solution, cooled, and set aside.

[0051] (3) Agarose activation: add 1.5 g of N,N carbonyldiimidazole (CDI) to the agarose solution in step (2), so that the concentration of CDI is 5% by mass to volume, activate at 25° C. for 1 hour, and obtain activated agarose solution.

[0052] (4) Agarose-grafted gelatin: mix the gelatin solution in step (1) with the activated agarose solution in step (3), react for 5 hours at 55°C in an oil bath, add deionized water to terminate the re...

Embodiment 2

[0058] Embodiment 2 skin tissue engineering scaffold

[0059] (1) Use PBS (0.01M, pH7.4) to dissolve the three kinds of agarose-gelatin graft temperature-sensitive hydrogels with grafting rates of 12.7%, 40.31% and 86.32% prepared in Example 1 at 100°C, so that The concentration is 3% by mass to volume, and it is sterilized and put into a thermos cup to keep warm. The fibroblast L929 in good growth condition was digested and prepared to a cell concentration of 2.4×10 7 cells / mL of cell suspension, take 50 μL of cell suspension and put it into a 8×8×2mm silica gel mold, cool the insulated agarose-gelatin graft temperature-sensitive hydrogel solution to 40°C and take 100 μL into the silica gel mold Stir quickly, take it out after it cools down to gel, put it into a 24-well plate, add 1mL DMEM complete medium for culture, and set fibroblast L929 as a control group added to agarose gel with a molecular weight of 50kDa. When cultured to 21 days, the survival rate of fibroblast L9...

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Abstract

The invention discloses an agarose-gelatin graft temperature-sensitive hydrogel, and a preparation method and application thereof. According to the invention, hydroxyl of degraded agarose is activatedby utilizing soluble N,N-carbonyldiimidazole, then the activated hydroxyl reacts with amino on gelatin to form an amide bond, and unreacted gelatin is removed by the characteristic that the gelatin is soluble at 37 DEG C. Biological activity and degradation performance of the agarose are improved by using the gelatin, and meanwhile, the gelatin is endowed with the gelatinization property by adopting agarose under a body temperature (37 DEG C), and immunogenicity of the material is reduced. The obtained agarose-gelatin graft temperature-sensitive hydrogel can be used for embedding cells and providing a three-dimensional hydrogel support suitable for a growth environment of the cells, and also can be used as a controlled-release system to load a gene protein drug for tissue repair or disease diagnosis and treatment. The preparation method of the agarose-gelatin graft temperature-sensitive hydrogel is simple, a cross-linking agent is not needed, and the prepared temperature-sensitive hydrogel has low immunogenicity, is easy to degrade and has good biological activity.

Description

technical field [0001] The invention relates to the field of medical technology, in particular to an agarose-gelatin graft temperature-sensitive hydrogel and a preparation method and application thereof. Background technique [0002] The hydrogel is easy to operate and can carry functional components such as cells and growth factors for tissue repair. Hydrogels have the following advantages: first, they provide a highly hydrated environment similar to soft tissue, and after implantation, their flexible properties reduce friction or mechanical irritation of surrounding tissues; second, they have a semipermeable membrane with interconnected pores that allow substances to transport, including providing nutrients to the loaded cells and transporting metabolic waste; 3. It can protect the packaged cells from the immune attack of the host immune system; 4. It can be easily functionalized or carry functional components to enhance the cells -Interaction of materials to enhance the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08J3/075C08G81/00A61K9/06A61K47/42A61K47/36A61L27/22A61L27/20A61L27/52A61L27/50C08L87/00
CPCA61K9/06A61K47/36A61K47/42A61L27/20A61L27/222A61L27/50A61L27/52C08G81/00C08J3/075C08J2387/00C08L5/12
Inventor 汤顺清黄秀英
Owner JINAN UNIVERSITY