Bionic bleeding stopping biological glue

A hemostatic material and protein technology, applied in the field of bionic hemostatic biological glue, can solve problems such as difficult dressing changes, wound adhesion, and inadaptability to emergency bleeding emergencies

Active Publication Date: 2017-12-15
OCEAN UNIV OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Existing hemostatic gauze, hemostatic bandages and other hemostatic materials have certain limitations in use; for example, they are easy to adhere to the wound, it is not easy to change the dressing, and they are powerless against infection and purulent wounds
There are also some hemostatic materials mainly including collagen, oxidized regenerated cellulose, alginate, etc., but most of them have slow onset of action and are not suitable for emergency bleeding.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1 protein EGFL sbp9 Methods for in vitro expression of repetitive sequences.

[0024] By analyzing the scallop foot transcriptome, the present invention discovered the protein Sbp9 with cell adhesion function, and found a protein with EGF repeat sequence, named EGFL sbp9 , its amino acid sequence is as follows:

[0025] APCGGSCKANEHCDMHSQECVCNTGYKLYKKACVLPCGGPCKQYERCDEGSNKCVCMTGYSLFKGSCVVPCGGPCGPNAYCDKNKNQCNCNKGYFTYHGVCALPCGGPCKQNANCDKNSNQCVCNKGYK EIGGVCAV;

[0026] 1. EGFL sbp9 expression

[0027] The gene fragment was connected to pET28a by restriction endonuclease, and transformed into Escherichia coli BL21(DE3) to obtain a recombinant expression system. Grow it to OD using LB medium 600 When the concentration is 0.6-0.8, add IPTG to make the final concentration 0.2mM, and induce overnight. The cells were collected by centrifugation and stored at -80°C.

[0028] 2. EGFL sbp9 separation and purification of

[0029] The cells were resuspended in PBS an...

Embodiment 2

[0030] The chemical synthesis of embodiment 2 sodium alginate graft Dopa (Dopa-SA)

[0031] A Dopa-modified sodium alginate derivative was synthesized and named as Dopa-SA.

[0032] Weigh 0.2 g of sodium alginate and dissolve it in 15 mL of 0.1 M MES buffer solution with pH 6.3, stir at room temperature until dissolved, it takes about 1 h, and the final concentration is 1% (w / v). After the sodium alginate is completely dissolved, add 0.1162g NHS and 0.2903g EDC, keep the molar ratio of EDC to sodium alginate at 1.5-2.5:1, and keep the molar ratio of NHS to sodium alginate at 1.5-2.5:1 , stirred well at room temperature for 1 h.

[0033] Add 5ml of MES buffer solution with pH 6, add 0.2872g of dopamine hydrochloride, keep the molar ratio of dopamine to sodium alginate at 1.5:1, stir and mix at room temperature. After the dopamine hydrochloride was dissolved in the sodium alginate solution, the measured pH of the solution was 6.0 and stirred slowly at room temperature for 24 h...

Embodiment 3

[0034] Embodiment 3: the preparation of gel

[0035] The modified Dopa-SA was prepared into a solution with a concentration of 2% (wt) in PBS solution, stirred and dissolved at 4°C for 24 hours. Then centrifuge at 6000g for 10min to remove air bubbles. Take 200μl sample and drop it on the cover glass, add the prepared EGFL in proportion (molar ratio 5-10:1) sbp9 Recombinant protein preparation gel. It was found that after adding EGFL sbp9 After 5min, a water-insoluble bioglue with certain adhesion properties was obtained ( figure 2 ).

[0036] 1. Ultrastructural observation of bioglue

[0037] Take 1ml of DopA-SA and put it on the glass slide, add EGFL according to the ratio (molar ratio 5-10:1) sbp9 , stir well. After gel formation, the hydrogel was transferred into a 24-well plate and placed at -80°C for 24 hours, and then the frozen sample was subjected to vacuum freeze-drying. The freeze-dried hydrogel is white and fluffy, and the freeze-dried sample is quickly cu...

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Abstract

The invention provides bionic bleeding stopping biological glue with an amino acid sequence of SEQ ID NO:1. The invention also provides a bleeding stopping material which contains a protein with an amino acid sequence of SEQ ID NO:1 and sodium alginate modified by Dopa. The invention provides a product with bleeding stopping performance, and the product is prepared from polypeptide with an amino acid sequence SEQ ID NO:1 and a sodium alginate derivative; and application results show that the protein with the amino acid sequence of SEQ ID NO:1 can be used for improving ultramicrostructures, and in vitro experiments prove that the composite has good bleeding stopping effect.

Description

technical field [0001] The invention belongs to the technical field of biological materials, and in particular relates to a bionic hemostatic biological glue. Background technique [0002] With the diversification of human activities and lifestyles, there are more and more accidents, and the death caused by excessive blood loss has attracted more and more attention. Minor trauma is not enough to be fatal, and if the blood loss reaches 30% or more of the whole body's blood in a short period of time, it will be life-threatening. In order to save the lives of the injured in time, high-efficiency hemostatic materials with fast hemostatic effect and excellent biocompatibility are essential. [0003] Existing hemostatic gauzes, hemostatic bandages and other hemostatic materials have certain limitations in use; for example, they are easy to adhere to the wound, it is not easy to change the dressing, and they are powerless to infection and suppurative wounds. There are also some h...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/435A61L24/10A61L24/08A61L24/00
CPCA61L24/001A61L24/08A61L24/108A61L2400/04C07K14/43504C08L89/00C08L5/04
Inventor 刘伟治律倩倩汪丹丹徐平平
Owner OCEAN UNIV OF CHINA
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