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Orthopedics-department adhesive based on mussel adhesive protein

A mussel mucin and adhesive technology, which is applied in the fields of regenerative medicine and biomedical materials, can solve problems such as poor adhesion performance

Active Publication Date: 2019-10-25
ZHEJIANG RISING BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Mussel mucin currently the only bioadhesive commercially available (not cosmetic or wound covering fluid) is CELL-TAK produced by BD Bioscience, which can be used for tissue adhesion, while genetic engineering products lack the processing of mussel foot glands The adhesive performance is obviously not as good as that of natural extract products

Method used

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  • Orthopedics-department adhesive based on mussel adhesive protein
  • Orthopedics-department adhesive based on mussel adhesive protein
  • Orthopedics-department adhesive based on mussel adhesive protein

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Preparation of Example 1 Adhesive Kit and Adhesive The preparation of modified mussel mucin

[0038] Use double-distilled water to prepare CELL-TAK natural mussel mucin mussels as a suspension with a concentration of 0.5%, and add D-galactosamine and 20U / g mussel mucin mass to the suspension For transglutaminase, add NaOH1M to adjust the pH to 7.0 / direct reaction (the pH of some batches of subsuspension is close to 7), react in a water bath at 40°C for 4 hours; then inactivate transglutaminase in a water bath at 80°C for 5 minutes; use The 1000Da membrane was dialyzed against double distilled water for 24 hours to remove excess D-galactosamine (change the medium 3 times); freeze-dried to obtain modified mussel mucin.

[0039] Adhesive preparation

[0040] The modified mussel mucin needs to be refrigerated separately at 4°C before use. When in use, it can be prepared as an adhesive with Tween 80 and BMP-2, which are also refrigerated at 4°C and 5% acetic acid solution p...

Embodiment 2

[0044] The adhesion performance test of embodiment 2 adhesive and contrast adhesive

[0045]Select 12 male New Zealand big-eared white rabbits weighing about 2.5 kg. Immediately after execution, take the complete ulna of the left forelimb, cut off the middle section vertically with an electric saw, and then use 60 μL of the adhesive prepared in Example 1, the control adhesive 1 and the control adhesive Mixture 2 product bonding fracture, press for 10 minutes and let it stand for 30 minutes for testing.

[0046] Test the mechanical properties with a mechanical tester (three-point bending test, acceleration 1mm / min, fulcrum distance 2cm), and take the average value of the three samples. The results are shown in Table 1.

[0047] Table 1 Bending strength of adhesive bonded bone specimens

[0048]

[0049]

[0050] It can be seen from the above table that the adhesive performance of mussel mucin obviously exceeds that of the existing fibrin glue. Fibrin glue.

Embodiment 3

[0051] The in vitro release experiment of embodiment 3BMP2

[0052] Obtain 0.5 mL each of the adhesive prepared in Example 1 and the reference adhesive 1, manually press it into a thin sheet with a diameter of about 1 cm, and place it in a dialysis bag after solidification for 1 hour. The dialysis bag is placed in 15 ml of 0.2% azide Sodium azide, pH 7.0 in PBS buffer; stand at 37°C, take 1ml of samples at 12h, 24h, 120h, 240h, 480h, 720h, and 960h (then add 0.2% sodium azide, pH7 .0 PBS buffer solution 1ml), according to the instructions of the kit (human BMP-2 ELISA detection kit, produced by Tianjin Anuo Ruikang Biotechnology Co., Ltd.), the BMP-2 concentration was measured by ELISA method, and the cumulative release percentage was calculated by conversion. The results are shown in Table 2:

[0053]

[0054] It can be seen that the addition of tween 80 promotes the formation of microstructures such as mussel mucin spherical sheets and plays a slow-release effect on BMP-...

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Abstract

The invention provides an orthopedics-department adhesive containing D-galactosamine glycosylation modified mussel adhesive protein, Tween 80 and BMP-2, an adhesive kit with various components packaged separately and application. The adhesive can slowly release the BMP-2 while the basic adhesive performance is kept to achieve a good bone fracture recovery promoting effect.

Description

technical field [0001] The invention belongs to the field of biomedical materials containing active protein and the field of regenerative medicine. Specifically, the application provides an orthopedic mucin containing D-galactosamine glycosylated mussel mucin, Tween 80, and BMP-2 Mixture, and provide the adhesive kit that each component of this adhesive is packaged separately and the application of this adhesive. Background technique [0002] Bone morphogenetic protein-2 (bone morphogenetic protein-2, BMP-2) is a key factor in promoting bone formation and belongs to the TGF-α family member. BMP-2 is the main signaling molecule for cells to differentiate into mineral deposition osteoblasts, and plays a role in inducing osteoblast differentiation. Expressed in limb growth, endochondral ossification, and fractures, it plays an important role in bone growth and regeneration and repair. It is considered to be a good pharmaceutical active ingredient for the treatment of bone dise...

Claims

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Application Information

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IPC IPC(8): A61L24/10A61L24/00A61K38/18A61P19/08A61P19/02
CPCA61K38/1841A61L24/0015A61L24/108A61L2300/252A61L2300/412A61L2300/602A61L2400/04A61P19/02A61P19/08C08L89/00
Inventor 陈春曹建新
Owner ZHEJIANG RISING BIOTECH CO LTD
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