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Preparation method of co-crosslinked double network hydrogel scaffold for promoting osteogenic growth

A co-crosslinking and double-network technology, applied in tissue regeneration, medical science, prosthesis, etc., can solve the problems of hydrogel strength decrease, drug release, etc., to promote bone density increase, increase bone formation, and enhance adsorption The effect of the aggregation function

Active Publication Date: 2017-09-29
THE FIRST AFFILIATED HOSPITAL OF SOOCHOW 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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  • Preparation method of co-crosslinked double network hydrogel scaffold for promoting osteogenic growth
  • Preparation method of co-crosslinked double network hydrogel scaffold for promoting osteogenic growth
  • Preparation method of co-crosslinked double network hydrogel scaffold for promoting osteogenic growth

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1 Preparation of Co-crosslinked Double Network Hydrogel Scaffold for Promoting Osteogenic Growth

[0045] The specific steps are:

[0046] 1) Preparation of methacrylic anhydride modified gelatin

[0047] figure 2 This is the flow chart for the preparation of methacrylic anhydride-modified gelatin. Add 200 mL of phosphate buffer to 20 g of gelatin, and keep stirring at 60°C for 2 hours; Add 1 mL of methacrylic anhydride to the gelatin mixture every 4 minutes while stirring, add 16 times in total, and keep stirring for 2 hours to form methacrylic anhydride-modified gelatin; 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

[0054] Example 2 Adhesion and Aggregation Observation of GelMA and GelMA-OGP

[0055] Experimental groups: control group, GelMA group, GelMA-OGP scaffold group.

[0056] Put the GelMA and GelMA-OGP scaffolds into 24-well plates first, then recover MC3T3-E1, centrifuge, count, and plant them on the scaffolds in two 24-well plates (2×10 per well) 4 Cells) (MC3T3-E1 in the control group were directly planted on the culture plate), three replicate wells in each group were added to the prepared α-MEM medium, and cultured in the incubator for 24 hours until the cells fully adhered to the wall. The α-MEM medium was changed every two days, and the control group only changed the α-MEM medium on the culture plate, and cultured for 1 day and 3 days respectively, and observed with live-death staining fluorescence microscope.

[0057] The procedure for live-dead staining is as follows: suck out the culture medium → wash twice with PBS → add 100 ul of the mixture to each well (1 μl of A +...

Embodiment 3

[0059] Example 3 Morphological observation of MC3T3-E1 implanted on the scaffold

[0060] Experimental groups: control group, GelMA group, GelMA-OGP scaffold group.

[0061] Put the GelMA and GelMA-OGP scaffolds into 96-well plates first, then resuscitate, centrifuge, count and plant MC3T3-E1 on the scaffolds in 96-well plates (2×10 per well 3 Cells) (control group MC3T3-E1 were directly planted on the culture plate), added α-MEM medium, and cultured in the incubator for 24 hours, until the cells fully adhered to the wall and grew. The α-MEM medium was replaced every other day (the control group only replaced the α-MEM medium on the culture plate). On the 3rd day, Dapi and phalloidin staining were performed for fluorescence microscope observation.

[0062] Dapi and phalloidin staining steps: Aspirate the culture medium → wash with PBS → fix with paraformaldehyde for 15 minutes → wash with PBS for 2 minutes*5 times → 1ul phalloidin + 300ul PBS (1:300) dilution → 96-well plat...

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Abstract

The invention belongs to the field of biomedical materials and in particular relates to a preparation method of a co-crosslinked double network hydrogel scaffold for promoting osteogenic growth. The preparation method comprises the following steps: (1) preparation of GelMA; (2) preparation of photo-crosslinkable OGP; and (3) preparation of the hydrogel scaffold: dissolving 50mg of the GelMA prepared in the step (1) and 20mg of the photo-crosslinkable OGP prepared in the step (2) in 1mL of PBS, uniformly mixing the two, adding 10mg of a light initiator, uniformly mixing the mixture, and irradiating the mixture by ultraviolet rays to prepare the hydrogel scaffold. The hydrogel scaffold prepared by the invention plays obvious roles of promoting increase of bone density and union of bone defects, and can regulate proliferation and differentiation of osteoblast and mineralization of a matrix and improve the alkaline phosphatase activities. The hydrogel scaffold is of a porous structure, so that OGP is co-crosslinked to the surface of the porous scaffold to induce OGP to slowly release, and therefore, actions of the biological scaffold in the fields of orthopedics department such as bone defects and bone graft fusion are achieved.

Description

technical field [0001] The invention belongs to the technical field of biomedical materials, and in particular relates to a preparation method of a co-crosslinked double network hydrogel scaffold for promoting osteogenic growth. Background technique [0002] According to literature reports, with the improvement of the economic level and the acceleration of industrialization, the number of cars has increased year after year, and more than 50,000 bone defects caused by high-energy injuries require surgical treatment every year. In some complex situations, such as large bone defects caused by tumor resection, osteoporosis, infection and trauma, the regeneration and homeostasis of bone is facing great challenges. Common methods used clinically for bone defect repair include allogeneic bone grafting, autologous bone grafting, and the use of BMP-2 factors. However, these three methods have various disadvantages, such as limited source of autologous bone, prolonged operation time,...

Claims

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Application Information

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IPC IPC(8): A61L27/48A61L27/52A61L27/54A61L27/56A61L27/50
CPCA61L27/48A61L27/50A61L27/52A61L27/54A61L27/56A61L2300/252A61L2300/412A61L2300/602A61L2430/02C08L89/00
Inventor 施勤崔文国潘国庆刘星志乔渝森孙智勇朱雪松
Owner THE FIRST AFFILIATED HOSPITAL OF SOOCHOW UNIV
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