An injectable in situ hydrogel imitating extracellular matrix and its preparation method and application

A technology of hydrogel and extracellular matrix, which is applied in the field of in situ hydrogel which imitates extracellular matrix and can be injected and its preparation and application, can solve the problem of single release rate, complicated steps, no controllable degradation, controllable Issues such as release and/or regulation of cell behavior, to achieve controllable release and/or regulation of cell behavior, to meet the requirements of different release rates, and to have broad clinical application prospects.

A technology of hydrogel and extracellular matrix, which is applied in the field of in situ hydrogel which imitates extracellular matrix and can be injected and its preparation and application, can solve the problem of single release rate, complicated steps, no controllable degradation, controllable Issues such as release and/or regulation of cell behavior, to achieve controllable release and/or regulation of cell behavior, to meet the requirements of different release rates, and to have broad clinical application prospects.

CN104307049BActive Publication Date: 2016-04-13WEST CHINA HOSPITAL SICHUAN UNIV
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  • An injectable in situ hydrogel imitating extracellular matrix and its preparation method and application
  • An injectable in situ hydrogel imitating extracellular matrix and its preparation method and application
  • An injectable in situ hydrogel imitating extracellular matrix and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0072] Example 1 Preparation of injectable in situ hydrogel imitating extracellular matrix

[0073] (1) Preparation of thiolated gelatin

[0074] Add 1 g of gelatin into 100 ml of ultrapure water, heat to 40°C, stir to dissolve, and then cool to room temperature. Cystamine dihydrochloride, EDC (1-(3-dimethylaminopropyl)-3-ethylcarbodiethylene amine hydrochloride) and NHS (N-hydroxysuccinimide), adjust the pH of the reaction solution to 4.75, start dialysis after 4 hours of reaction at room temperature, change the dialysate every 24 hours, add 0.5gDTT (dithiothreose) after 3 days of dialysis Alcohol), adjust the pH of the reaction solution to 8.5, adjust the pH to 4.0 after 2 hours of reaction, dialyze under nitrogen protection for 3 days, and change the dialysate once a day. After the dialysis is completed, filter sterilization and freeze-dry to obtain thiolated gelatin. The schematic diagram of its preparation is as follows: figure 1 .

[0075] The thiol content was deter...

Embodiment 2

[0084] Example 2: Preparation of injectable in situ hydrogel imitating extracellular matrix

[0085] (1) Preparation of thiolated gelatin

[0086] Add 1 g of gelatin into 100 ml of ultrapure water, heat to 40°C, stir to dissolve, and then cool to room temperature. Add cystamine dihydrochloride, EDC and NHS according to the molar ratio of 1:1:2:2 (the molar ratio of carboxyl groups in gelatin is 1), adjust the pH of the reaction solution to 4.75, and start dialysis after 4 hours of reaction at room temperature, every 24 hours Change the dialysate once, add 0.5g DTT after 3 days of dialysis, adjust the pH of the reaction solution to 8.5, react for 2 hours, adjust the pH to 4.0, dialyze for 3 days under nitrogen protection, and change the dialysate once a day. After the dialysis is completed, filter sterilization and freeze-drying to obtain thiolated gelatin.

[0087]The mercapto group content was determined to be 0.42 mmol / g by the Ellman method.

[0088] (2) Preparation of t...

Embodiment 3

[0096] Example 3: Preparation of injectable in situ hydrogel imitating extracellular matrix

[0097] (1) Preparation of thiolated gelatin

[0098] Add 1 g of gelatin into 100 ml of ultrapure water, heat to 40°C, stir to dissolve, and then cool to room temperature. Add cystamine dihydrochloride, EDC and NHS according to the molar ratio of 1:3:3:3 (the carboxyl molar ratio in gelatin is 1), adjust the pH of the reaction solution to 4.75, and start dialysis after 4 hours of reaction at room temperature, every 24 hours Change the dialysate once, add 0.5g DTT after 3 days of dialysis, adjust the pH of the reaction solution to 8.5, react for 2 hours, adjust the pH to 4.0, dialyze for 3 days under nitrogen protection, and change the dialysate once a day. After the dialysis is completed, filter sterilization and freeze-drying to obtain thiolated gelatin.

[0099] The mercapto group content was determined to be 0.6 mmol / g by the Ellman method.

[0100] (2) Preparation of mercaptohep...

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Abstract

The invention discloses imitated extracellular matrix injectable in-situ hydrogel and a preparation method and application thereof. The hydrogel is prepared by carrying out a cross-linking reaction between sulfhydrylation gelatin, sulfhydrylation polysaccharides or RGD-containing cell adhesion peptides and polyethylene glycol diacrylate. The composition of the hydrogel is similar to that of the extracellular matrix, so that the hydrogel has high biocompatibility, has the effects of controlled degradation, controlled release and / or cell behavior regulation in performance and can conveniently meet the requirements on different release rates of growth factors or medicines. Meanwhile, the clinical injectable and in-situ forming operation requirements can be met, and the hydrogel has good application prospects in tissue repair and regeneration.

Description

technical field [0001] The invention relates to an injectable in-situ hydrogel imitating extracellular matrix, a preparation method and application thereof. Background technique [0002] For a long time, human beings have been exploring the repair and regeneration of body tissues. In recent years, with the development of biomaterials, tissue engineering and stem cell technology, breakthroughs have been made in the field of tissue repair and regeneration. Current research mainly focuses on two aspects: one is to induce endogenous cells to perform tissue reconstruction in the diseased site by administering drug treatment such as growth factors; the other is to implant cells to play a therapeutic role in the diseased site. In both strategies, the design and development of carrier materials undoubtedly plays an increasingly important role. [0003] First, the degradation of the carrier material should match the tissue growth. Secondly, the carrier used for drug delivery must ...

Claims

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Application Information

Patent Timeline
13 Apr 2016
Publication
CN104307049B
IPC
A61K47/42; A61K47/36; A61L27/52; A61L27/26; A61L27/54; A61K47/34; A61K38/18; A61K9/08; C12N5/07; A61L27/38; A61K35/12
CPC
A61L27/26; A61L27/52; A61L27/54; A61L2300/252; A61L2300/412; A61L2430/02; C08L5/04; C08L5/08
Inventors
田猛; 游潮