RhBMP-2-loaded chitosan microspheres, preparation method thereof and application thereof

A technology of chitosan microspheres and rhbmp-2, which is applied in the direction of pharmaceutical formulations, medical preparations containing no active ingredients, medical preparations containing active ingredients, etc., can solve the problems of short half-life, easy degradation by enzymes, and bioavailability Low-level problems, to achieve the effect of promoting new bone formation, simple operation, high adsorption capacity

Inactive Publication Date: 2011-08-03
EAST CHINA UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, pure recombinant human bone morphogenetic protein-2, like other cell growth factors, is easily degraded by enzymes and has a short half-life in vivo. It is quickly diluted and metabolized in local and systemic applications, and its bioavailability is very low. Therefore, recombinant human bone morphogenetic protein-2 2 requires a suitable carrier for effective play

Method used

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  • RhBMP-2-loaded chitosan microspheres, preparation method thereof and application thereof
  • RhBMP-2-loaded chitosan microspheres, preparation method thereof and application thereof
  • RhBMP-2-loaded chitosan microspheres, preparation method thereof and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] The molecular weight is 30KDa, the chitosan that deacetylation degree is 90%, is dissolved in 2% (v / v) acetic acid solution, is mixed with 4 (w / v)% chitosan solution, slowly adds into 5 Double the volume of oil phase liquid paraffin containing surfactant (the volume ratio of Span-80 and Tween-80 is 2:1, and the volume ratio of surfactant and oil phase is 1:100.), stir for 3h to form W / O emulsion, add an equal volume of 20% (w / w) sulfuric acid solution for cross-linking, continue stirring for 4 hours after cross-linking, add acetone for dehydration, stand to separate layers, and wash the precipitate with acetone and absolute ethanol distilled water respectively. Then the precipitate is dispersed in water, and a sodium hydroxide solution with a concentration of 4% (w / v) is added dropwise under magnetic stirring until the final pH value of the reaction system is greater than 7, then fully washed with acetone, absolute ethanol and water, and frozen Chitosan microspheres ar...

Embodiment 2

[0038] Be that the chitosan that molecular weight is 50.5KDa, degree of deacetylation is 90%, be dissolved in 3% (v / v) acetic acid solution, be mixed with 3% chitosan solution after defoaming, slowly add into 7 times volume containing In the oil phase liquid paraffin of surfactant Span-80 (the volume ratio of surfactant and oil phase is 1:100), stir for 4 hours to form a W / O emulsion, add 2 times the volume of 10% (w / w) sulfuric acid The solution was cross-linked, and after stirring and cross-linking for 6 hours, acetone was added for dehydration, the layers were left to stand, and the precipitates were washed with acetone and absolute ethanol distilled water respectively. Precipitation is dispersed in water then, under magnetic stirring, dropwise concentration is the sodium hydroxide solution of 2% (w / v), until the final pH value of reaction system is greater than 7; w) Soak in sulfuric acid aqueous solution for 1 minute, then fully wash with acetone, absolute ethanol and wat...

Embodiment 3

[0041] Dissolve chitosan with a molecular weight of 50KDa and a deacetylation degree of 95% in 2% (v / v) acetic acid solution, prepare a 3% chitosan solution for defoaming, and slowly add it to 3 times the volume of the surface-containing Active agent Tween-80 (the volume ratio of surfactant and oil phase is 2:100). In the oil phase liquid paraffin, stir for 2 hours to form a W / O emulsion, add an equal volume of 30% (w / w) sulfuric acid solution for cross-linking, continue stirring and cross-linking for 1 hour, add acetone for dehydration, and let stand to separate layers. The precipitate was washed with acetone and absolute ethanol with distilled water, respectively. Then the precipitate is dispersed in water, and a sodium hydroxide solution with a concentration of 4% (w / v) is added dropwise under magnetic stirring until the final pH value of the reaction system is greater than 7; then fully washed with acetone, absolute ethanol and water, frozen Chitosan microspheres are obta...

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Abstract

The invention discloses recombinant human bone morphogenetic protein-2 (rhBMP-2)-loaded chitosan microspheres, a preparation method thereof and application thereof. A substrate of the rhBMP-2-loaded chitosan microspheres is chitosan microspheres, and a loaded medicament is rhBMP-2. Chitosan reacts with an ionic cross-linking agent, and a reaction product is processed by solution of sodium hydroxide to prepare the chitosan microspheres; and the rhBMP-2 is loaded by a passive loading mode, so that the rhBMP-2 is adsorbed on the chitosan microspheres, wherein a weight ratio of the rhBMP-2 to thechitosan is (1-30):100. The preparation method is simple and easy, has a mild preparation condition and is favorable for maintaining the bioactivity of the rhBMP-2. The rhBMP-2-loaded chitosan microspheres can control and release the rhBMP-2 for a long term, promote the formation of new bones, and can be used for preparing medicaments for treating bone coloboma, nonunion and delayed union.

Description

technical field [0001] The invention relates to a protein bioactive factor carrier and a preparation method thereof, in particular to a chitosan microsphere loaded with recombinant human bone morphogenetic protein-2 (rhBMP-2) and a preparation method thereof. Background technique [0002] With the development of genetic engineering and biotechnology, more and more biologically active factors are used to treat and prevent diseases. Bone morphogenetic protein-2 (BMP-2), a member of the transforming growth factor β superfamily, has the ability to induce undifferentiated mesenchymal stem cells to differentiate and proliferate toward chondrocytes and osteoblasts. ability, plays an important role in repairing bone defects and promoting bone healing (Reddi, A.H., Bone morphogenetic proteins: from basic science to clinical applications. J Bone Joint Surg Am, 2001, 83-A Suppl 1 (Pt 1): S1- 6). However, pure recombinant human bone morphogenetic protein-2, like other cell growth fact...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K9/16A61K38/18A61K47/36A61P19/08
Inventor 刘昌胜钱秀珍王靖
Owner EAST CHINA UNIV OF SCI & TECH
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