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Preparation method and application of gelatin microspheres

A technology of gelatin and microspheres, which is applied in the field of regenerative medicine, can solve the problems of aggravated microsphere adhesion, easy adhesion, and low sphericity, and achieve the effect of simple operation, low cost, and reduced adhesion

Inactive Publication Date: 2019-05-24
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the cross-linking process of genipin is slower than that of glutaraldehyde, which usually takes 12-72 hours, while glutaraldehyde only takes 1-2 hours. The surface is not fully cross-linked and sticks to each other, which leads to lower sphericity and poorer dispersion of genipin cross-linked microspheres than glutaraldehyde cross-linked microspheres
The current conventional preparation method of genipin cross-linked microspheres is to immerse uncross-linked microspheres in genipin solution for cross-linking after lyophilization. However, uncross-linked gelatin microspheres are prone to adhesion during freeze-drying, and The cross-linking reaction intensified the adhesion between microspheres under static conditions, which resulted in a lot of cohesive microspheres after cross-linking
Although some researchers added genipin solution for cross-linking when stirring to form water-in-oil droplets, the droplets collided with each other under stirring conditions, and the slower cross-linking speed of genipin still produced more cohesive microparticles. ball

Method used

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  • Preparation method and application of gelatin microspheres
  • Preparation method and application of gelatin microspheres
  • Preparation method and application of gelatin microspheres

Examples

Experimental program
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Embodiment 1

[0037] Prepare 20 g of 25% (w / v, g / ml) alkaline gelatin (120 Bloom g) aqueous solution and preheat it in a water bath at 53° C. for 1.5 h. In addition, prepare liquid paraffin containing span80 (the density of liquid paraffin is 0.9, and the volume ratio of liquid paraffin to span80 is 50:1), mix well, and preheat in a water bath at 53°C for 1.5h. After that, the gelatin solution was slowly added dropwise to the liquid paraffin under the stirring condition of 1200rpm, the volume ratio of the aqueous gelatin solution to the liquid paraffin containing span80 was 1:10, and the stirring time was 15min to prepare stable water-in-oil droplets. Then, under the condition of constant stirring speed, add an ice bath to cool down, and stir for 30 minutes to form solidified microspheres. Afterwards, 5% (w / v, g / ml) genipin solution was added, the final concentration of which was 0.3% (w / v, g / ml) when cross-linked, and the reaction was stirred for 72 hours. Then, slowly add isopropanol wit...

Embodiment 2

[0039] Prepare 20 g of an aqueous solution of 10% (w / v, g / ml) acid gelatin (180 Bloom g), and preheat it in a water bath at 53° C. for 2 h. In addition, prepare liquid paraffin containing span80 (density specific gravity of liquid paraffin is 0.89, volume ratio of liquid paraffin and span80 is 30:1), mix well, and preheat in 53°C water bath for 2h. After that, the gelatin solution was slowly added dropwise to the liquid paraffin under the stirring condition of 700rpm, the volume ratio of the aqueous gelatin solution to the liquid paraffin containing span80 was 1:10, and the stirring time was 9min to prepare stable water-in-oil droplets. Then, under the condition of constant stirring speed, add an ice bath to cool down, and stir for 15 minutes to form solidified microspheres. Afterwards, 2% (w / v, g / ml) genipin solution was added, the final concentration of which was 0.03% (w / v, g / ml) when cross-linked, and the reaction was stirred for 12 hours. Then, slowly add isopropanol wit...

Embodiment 3

[0041]Prepare 30 g of an aqueous solution of 18% (w / v, g / ml) alkaline gelatin (200 Bloom g), and preheat it in a water bath at 53° C. for 2.5 h. In addition, prepare the liquid paraffin containing span80 (the density of liquid paraffin is 0.9, the volume ratio of liquid paraffin and span80 is 40:1), mix well, the volume ratio of gelatin aqueous solution and the liquid paraffin containing span80 is 1:10, and in Preheat in a water bath at 53°C for 2.5h. After that, the gelatin solution was slowly added dropwise into the liquid paraffin under the stirring condition of 1000rpm, and the stirring time was 12min to prepare stable water-in-oil droplets. Then, under the condition of constant stirring speed, add an ice bath to cool down, and stir for 20 minutes to form solidified microspheres. Afterwards, add 5% (w / v, g / ml) genipin solution, its final concentration is respectively 0.02% (w / v, g / ml), 0.03% (w / v, g / ml) when cross-linking , 0.3% (w / v, g / ml) and 0.4% (w / v, g / ml), stirred ...

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Abstract

The invention provides a preparation method and application of gelatin microspheres. The preparation method includes steps: (1), dropwise adding a preheated gelatin solution into preheated liquid paraffin containing span 80 while stirring to prepare stable water-in-oil liquid drops; (2), cooling the water-in-oil liquid drops in ice bath to form cured microspheres; (3), adding in a genipin solutionfor crosslinking; (4), adding in isopropanol for demulsification, applying isopropanol and purified water for centrifugal washing, and freeze-drying to finally obtain dried gelatin microspheres. Thegelatin microspheres are high in sphericity and dispersity, low in cytotoxicity, of a netty structure and capable of adsorbing and slowly releasing growth factor and can be used for in-vivo injectiontherapy.

Description

technical field [0001] The invention relates to the field of regenerative medicine, in particular to a preparation method of slow-release growth factor gelatin microspheres. Background technique [0002] Gelatin microspheres are an injection carrier. Since gelatin molecules are charged, they can adsorb growth factors through electrostatic interaction. Therefore, gelatin microspheres can carry growth factors and inject them into damaged parts of the body, and release growth factors in situ slowly. application prospects. Uncrosslinked gelatin microspheres have low strength and degrade too quickly in vivo, while crosslinked microspheres have good strength and long degradation time, which can meet the needs of long-term in situ growth factor release in vivo. Glutaraldehyde is a widely used cross-linking agent. Glutaraldehyde cross-linked gelatin microspheres have short preparation time, high sphericity and good dispersibility, but glutaraldehyde is cytotoxic, although glycine c...

Claims

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Application Information

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IPC IPC(8): A61K9/19A61K38/18
Inventor 孙广炜刘洋张英赵姗
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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