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Vascular endothelial growth factor chitosan microsphere and application thereof

A technology of chitosan microspheres and vascular endothelium, applied in the field of biomedicine, can solve problems such as protein cytokine damage, and achieve the effects of promoting fracture healing, mild reaction conditions, and speeding up the osteogenesis process.

Inactive Publication Date: 2011-06-22
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the use of chemical cross-linking agents such as glutaraldehyde or formaldehyde, it has a certain destructive effect on protein cytokines, so its application has certain limitations.

Method used

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  • Vascular endothelial growth factor chitosan microsphere and application thereof
  • Vascular endothelial growth factor chitosan microsphere and application thereof
  • Vascular endothelial growth factor chitosan microsphere and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] The modified Berthold method was used to prepare chitosan microspheres. The method was as follows: Dissolve 200 mg chitosan in 80 ml 2% glacial acetic acid, add 1 ml Tween-80 magnetically stirred for 2 hours, and ultrasonically oscillate until the solution was clear. Under magnetic stirring at room temperature, 20 ml of cross-linking agent sodium polyphosphate with a concentration of 2.5 mg / ml was slowly added dropwise, and stirred magnetically for 2 hours. 10000rpm / min, centrifuged for 15 minutes, and the resulting precipitated pure water was repeatedly centrifuged and washed 3 times, collected and sent to freeze-drying for storage to obtain blank chitosan microspheres. The concentration of chitosan and cross-linking agent sodium polyphosphate is 2.5g / L, and the volume ratio is 4:1.

[0017] Preparation of VEGF chitosan microspheres: Accurately weigh 20 mg of chitosan freeze-dried microspheres into an EP tube, add 0.9ml 2% glacial acetic acid and 0.1ml VEGF, and oscill...

Embodiment 2

[0018] Example 2 Encapsulation Efficiency and Drug Loading Research

[0019] Get the obtained centrifugate (1ml) of Example 1, each washing supernatant (1ml), dilute 50 times with PBS, get 1ml in the centrifuge tube as the sample to be tested, operate according to the ELISA kit instructions, measure its absorbance (A450 )value. Calculate the concentration of VEGF in the sample according to the standard equation of the standard curve, thereby deduce the total amount of VEGF in the centrifugate, washing supernatant and combined solution.

[0020] Table 1 Determination of VEGF concentration in samples

[0021]

[0022] Table 2 Determination of VEGF concentration in standard solution

[0023] According to Table 2, the linear equation of the standard solution concentration is obtained: y = 0.0014x + 0.1477 (x is the concentration, y is the absorbance value), calculate the corresponding concentration (see Table 1), parallelize three groups of experiments, and combine the ce...

Embodiment 3

[0028] Example 3 In vitro drug release research of VEGF chitosan microspheres

[0029] Weigh 20 mg of VEGF-chitosan microspheres into a centrifuge tube, add 1ml PBS, mix well at 37°C, and centrifuge at 15,000 r / min at 1, 5, 10 h and 1, 2, 3, 5, and 7 d respectively After 10 min, take 150ul of the supernatant and add 150ul of PBS to continue mixing. The collected supernatant samples were stored at -20°C. Each sample was used as the sample to be tested, and the A450 value was determined according to the kit instructions, and the corresponding VEGF concentration was calculated to calculate the VEGF content in the sample.

[0030] Table 3 Determination of VEGF concentration at different times

[0031] .

[0032] Table 4 Determination of VEGF concentration in standard solution

[0033]

[0034] According to Table 4, the standard solution concentration linear equation is obtained: y = 0.0014x + 0.1131 (x is the concentration, y is the absorbance value), calculate the corr...

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Abstract

The invention provides a vascular endothelial growth factor chitosan microsphere. The matrix of the microsphere is chitosan with deacetylation degree of more than or equal to 90.0 percent and viscosity of 100mPa.s, the coated active protein is a vascular endothelial growth factor, and the microsphere has the grain size of 200 to 300 nanometers. The chitosan microsphere serves as a carrier, the vascular endothelial growth factor is coated, and experiments prove that: the chitosan microsphere can well encapsulate and envelop the vascular endothelial growth factor and can well control the release of the vascular endothelial growth factor in a longer time. Moreover, the carrier is not needed to be taken out after administration and can be completely degraded and metabolized in vivo, decomposition products and metabolic products of the carrier are harmless to a human body, and the chitosan microsphere can be applied in preparing medicaments for promoting vascular endothelial cell proliferation and promoting growth of new blood vessels.

Description

technical field [0001] The invention belongs to the field of biomedicine, and specifically relates to a vascular endothelial growth factor chitosan microsphere, which can carry vascular endothelial growth factor and control its release, and can be used in the preparation of vascular endothelial cell proliferation and Application of drugs for promoting neovascularization. Background technique [0002] Vascular endothelial growth factor (VEGF) is a glycoprotein that widely exists in tissue cells and has various biological functions. VEGF has a strong function of promoting the proliferation of vascular endothelial cells and the growth of new blood vessels. By enhancing the expression of VEGF, VEGF and its gene therapy can improve myocardial ischemia, resist atherosclerosis, prevent and treat arteriovenous vessel injury and vascular restenosis, promote cardiovascular transplantation, treat orthopedic ischemic diseases and fractures, etc. have an important role. If the express...

Claims

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Application Information

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IPC IPC(8): A61K9/16A61K38/19A61K47/36A61P17/02A61P19/08A61P43/00
Inventor 谈伟强杨虎宣贵达陈强黄建新
Owner ZHEJIANG UNIV
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