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A preparation method of co-crosslinked double network hydrogel scaffold for promoting cartilage damage repair

A cartilage injury and co-crosslinking technology, applied in the field of biomedical materials, can solve the problems of decreased hydrogel strength and fast drug release, and achieve the effects of promoting adhesion, promoting cartilage defect healing, and enhancing adsorption and aggregation functions.

Active Publication Date: 2020-06-23
SUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this type of hydrogel material has problems such as fast drug release and decreased hydrogel strength in body fluids.

Method used

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  • A preparation method of co-crosslinked double network hydrogel scaffold for promoting cartilage damage repair
  • A preparation method of co-crosslinked double network hydrogel scaffold for promoting cartilage damage repair
  • A preparation method of co-crosslinked double network hydrogel scaffold for promoting cartilage damage repair

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Example 1 Preparation of Co-crosslinked Double Network Hydrogel Scaffold for Cartilage Damage Repair

[0050] The specific steps are:

[0051] 1) Preparation of methacrylate-modified gelatin

[0052] figure 2 This is the flow chart for the preparation of methacrylate-modified gelatin. Add 200 mL of phosphate buffer to 20 g of gelatin, and keep stirring at 60°C for 2 hours; Add 1mL methacrylic anhydride to the gelatin mixture every 4 minutes while stirring, add 16 times in total, and keep stirring for 2 hours to form methacrylate-modified gelatin; the prepared methacrylate-modified Place the gelatin solution in 800mL preheated phosphate buffer solution for dilution, and continue to stir slowly for 15 minutes, place the diluted solution in a dialysis bag, the molecular weight cut-off of the dialysis bag is 10KD, soak in pure aqueous solution, Change the dialysate twice a day to remove unreacted methacrylic anhydride, and continue dialysis for one week; after dialysis,...

Embodiment 2

[0059] Example 2 GelMA, GelMA-HSNGLPL biocompatibility detection and observation

[0060] Experimental grouping: GelMA scaffold group, GelMA-HSNGLPL scaffold group.

[0061] Put the GelMA and GelMA-HSNGLPL scaffolds into 96-well plates first, then resuspend hMSCs, count them, and plant them on the scaffolds in two sets of 96-well plates (2×10 per well 3 Cells), five replicate wells in each group, add the prepared DMEM-L medium, at 37°C, 5% CO 2 They were cultured in an incubator, and the DMEM-L medium was replaced every two days, and the cells were cultured for 5 days to stain live and dead cells and observe under a fluorescent microscope.

[0062] The staining steps for living and dead cells are: suck out the culture medium → wash twice with PBS → add 50 ul of the mixture to each well (1μL A + 1μL B + 2mL PBS to form the mixture) → place in the incubator for 30 minutes → under the fluorescence microscope Observed.

[0063] After the cells were stained in the dark for 30 mi...

Embodiment 3

[0064] Example 3 Detection of hMSCs proliferation activity

[0065] Experimental groups: control group, GelMA stent group, GelMA-HSNGLPL stent group.

[0066] Resuspend hMSCs and seed into 96-well plates (3 plates, 2×10 per well 3 Cells), add DMEM-L medium or GelMA group or GelMA-HSNGLPL group leaching solution, and culture in an incubator until the cells fully adhere to the wall and grow. The 3 plates were tested for CCK-8 on the 1st, 3rd, 5th, and 7th day after cell attachment.

[0067] CCK-8 detection steps: Add 10ul CCK-8 reaction solution to the wells of the 96-well plate (3 groups, 3 replicate wells in each group) (avoid air bubbles in the wells) → incubate in an incubator for 2 hours → use a microplate reader Measure the absorbance (450nm).

[0068] CCK-8 detected the effect of GelMA and GelMA-HSNGLPL materials on the proliferation of hMSCs. The results show( Figure 7 ), GelMA, GelMA-HSNGLPL material leachate did not affect the growth of cells, and the material h...

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Abstract

The invention belongs to the field of biomedical materials and particularly relates to a preparation method of a co-crosslinked double-network hydrogel support for promoting cartilage repair and growth. The method comprises the following steps of (1) preparation of GelMA; (2) preparation of a photo-crosslinked polypeptide; and (3) preparation of a hydrogel support, namely preparing the hydrogel support from the GelMA and the photo-crosslinked HSNGLPL under the action of a photoinitiator. The prepared hydrogel support has an obvious effect of promoting new cartilage generation and cartilage defect healing, and is capable of regulating cartilage matrix secretion of normal chondrocytes around the injured tissue, mesenchymal stem cell migration to the injured part and cartilage differentiation; by virtue of a porous structure of the hydrogel support, the HSNGLPL can be crosslinked to the surface of a porous support, endogenous TFG-beta is effectively adsorbed, the concentration of local TFG-beta beta1 is improved and the effects of the biological support in the orthopedics fields, such as a cartilage defect and a cartilage injury are achieved.

Description

technical field [0001] The invention belongs to the technical field of biomedical materials, and in particular relates to a method for preparing a co-crosslinked double network hydrogel scaffold for promoting the repair of cartilage damage. Background technique [0002] According to literature reports, with the development of the economy and the improvement of the awareness of maintaining health through exercise, the number of cartilage defect operations caused by damage to articular cartilage tissue and other cartilage tissue diseases has increased sharply, such as microfractures, osteochondral transplantation, chondrocyte transplantation, and infection and The cartilage defect caused by trauma, the lack of blood vessels, nerves and other physiological characteristics of cartilage itself lead to great challenges in the balance of cartilage regeneration and self-repair after cartilage injury. Common methods used clinically for cartilage defect repair include joint replacemen...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K38/09A61L27/26A61L27/50A61L27/52A61L27/56A61L27/54C08H1/00C08J3/24C08L89/06
CPCA61L27/26A61L27/50A61L27/52A61L27/54A61L27/56A61L2300/252A61L2300/412A61L2300/602A61L2430/06C08H1/00C08J3/24C08J2389/06C08L89/00
Inventor 施勤潘国庆崔文国鞠晓晶顾巧丽
Owner SUZHOU UNIV
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