Method for preparing fibrin ferment-chitosan self-assembly nano particle and use thereof

A nanoparticle and thrombin technology, applied in the field of biomedicine, can solve problems such as poor stability, and achieve the effects of high mechanical strength, rapid hemostasis, and improved hemostasis effect.

Active Publication Date: 2009-07-22
GUANGZHOU BIOSEAL BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] In order to solve the above problems, the object of the present invention is to overcome the relatively poor stability of thrombin as a local hemostatic drug, and bring into play the multiple functions of hemostasis, film formation, drug carrier, biocompatibility and biodegradability of chitosan, Thereby providing a preparation method of thrombin-chitosan self-assembled nanoparticles with rapid hemostasis, adhesion, tissue sealing, quick and convenient use

Method used

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  • Method for preparing fibrin ferment-chitosan self-assembly nano particle and use thereof
  • Method for preparing fibrin ferment-chitosan self-assembly nano particle and use thereof
  • Method for preparing fibrin ferment-chitosan self-assembly nano particle and use thereof

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Embodiment 1

[0038] Get 20g of chitosan powder, put it in a three-necked flask, add 1000ml of 2% acetic acid aqueous solution, stir to make it dissolve completely, and form a 2.0% chitosan acetic acid aqueous solution; take 5g of thrombin (equivalent to 30000u) , add phosphate buffer (pH7.4) 100ml, stir and dissolve, form the buffer solution of thrombin; Slowly stir at room temperature, the buffer solution of thrombin is added dropwise in the above-mentioned chitosan acetic acid aqueous solution, and the time for adding is 30min, heat up to 40°C, add 30ml of 0.1% glutaraldehyde solution dropwise, stir continuously at a speed of 200 rpm for 30min, stop stirring, pour it into 1500°C liquid paraffin, stir for 25min, add 150ml of Tween 80 , stirred at a high speed of 500 rpm for 4 hours for emulsification; took the emulsion and added 180ml of 25% glutaraldehyde solution, stirred for 30min to make it evenly dispersed, and placed it in a water bath at 40°C for 8h of cross-linking and self-assembl...

Embodiment 2

[0040] Take 40g of chitosan powder, put it in a three-necked flask, add 1500ml of 2.5% acetic acid aqueous solution, stir to make it dissolve completely, and form a 2.6% chitosan acetic acid aqueous solution; take another 12g of thrombin (equivalent to 72000u) , add phosphate buffer (pH7.2) 200ml, stir and dissolve, form the buffer solution of thrombin; Slowly stir at room temperature, the buffer solution of thrombin is added dropwise in the above-mentioned chitosan acetic acid aqueous solution, and the time of dropping is 50min, heat up to 45°C, add 100ml of 0.5% polyethylene glycol solution dropwise, stir continuously at a speed of 150 rpm for 40min, stop stirring, pour it into 2500°C liquid paraffin, stir for 35min, add 200ml of Span 80, stir at a high speed of 800 rpm for 3 hours for emulsification; take the emulsion and add 300ml of 15% polyethylene glycol solution, stir for 40min to make it evenly dispersed, and place it in a water bath at 45°C for cross-linking and self-...

Embodiment 3

[0042] Get 30g of chitosan powder, put it in a three-necked flask, add 1500ml of 1% acetic acid aqueous solution, stir to make it dissolve completely, and form a chitosan acetic acid aqueous solution with a concentration of 2.0%; in addition, take 6g of thrombin (equivalent to 36000u) , add 150ml of oxalate buffer solution (pH7.5), stir and dissolve to form a buffer solution of thrombin; slowly stir at room temperature, add the buffer solution of thrombin dropwise to the above-mentioned chitosan acetic acid aqueous solution, and the time for adding For 40 minutes, heat up to 50°C, add dropwise 30ml of 0.5% sodium trethalate solution, stir continuously at a speed of 250 rpm for 20min, stop stirring, pour it into 2500°C liquid paraffin, stir for 25min, add 150g of paraffin For Loxamer, stir at a high speed of 800 rpm for 3 hours for emulsification; take the emulsion and add 250ml of 15% sodium thalassic acid solution, stir for 50min to disperse evenly, and place it in a water bat...

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Abstract

The invention relates to a method for preparing thrombin-chitosan self-assembly nano particles and application thereof. The method comprises: adding chitosan into an acetic acid aqueous solution to form a chitosan solution; adding thrombin into a buffer solution to form a thrombin solution; uniformly mixing the thrombin solution and the chitosan solution, adding a crosslinking agent for stirring, pouring the mixture into liquid paraffin for stirring, adding a dispersant, and performing emulsification; and adding the crosslinking agent for crosslinking reaction through water bath, performing filtration and acetone washing, using ethanol for washing and dehydration, and finally performing vacuum drying to obtain the thrombin-chitosan self-assembly nano particles. The method feeds the thrombin and the chitosan according to proportion to prepare thrombin-chitosan which is self-assembled into nano particles for the first time, maintains the stability of the thrombin of the thrombin-chitosan self-assembly nano particles, and gives better play to the comprehensive hemostatic effect of the thrombin and the chitosan of the thrombin-chitosan self-assembly nano particles. The method can be widely applied to tissue hemostasis, tissue sealing and tissue adhesion in the surgical fields such as general surgery, orthopaedics, heat and chest surgery, neurosurgery and gynaecology and obstetrics.

Description

technical field [0001] The invention belongs to the field of biomedicine, and relates to a preparation method and application of thrombin-chitosan self-assembled nanoparticles; specifically, it relates to a drug compound that can be used quickly and has the effects of rapidly stopping bleeding, bonding and sealing wounds , is a biological hemostatic material. Background technique [0002] Excessive blood loss is the main cause of death from accidents and war injuries, and it is also one of the main causes of death during surgical treatment. Control of blood loss is a key link in surgery, first aid and field trauma care. Unfortunately, bleeding (bleeding) from various causes is very common, accidental injury bleeding (bleeding) in daily life and work, traumatic bleeding (bleeding) in surgery, traumatic bleeding (bleeding) from war, etc., If it cannot be controlled in time, it may cause huge losses or even death. Therefore, hemostasis has increasingly become a research hotsp...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L15/38A61L15/28A61L24/10A61L24/08
Inventor 陈彥文李吉来肖尚志黄星江彩霞潘小宁
Owner GUANGZHOU BIOSEAL BIOTECH
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