A gene-activated antibacterial bioactive bone repair material and its preparation method and application
A biologically active and bone repairing technology, applied in the direction of antibacterial drugs, gene therapy, drug combination, etc., can solve the problems that non-viral vectors have not been reported, to improve cell compatibility and transfection rate, the preparation method is simple, The effect of easy operation
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[0035] A method for preparing a gene-activated antibacterial bioactive bone repair material of the present invention comprises the following steps:
[0036] 1) Add glycerin to chloroform first, stir and dissolve at room temperature, then add triethylamine, then drop acryloyl chloride into the mixture, and the molar ratio of the four is 1:(3~5):(3~5):( 3~5). Then the reaction temperature was increased to 45-55°C and stirred for 40-60 hours. After the reaction is complete, the mixture is washed with different reagents in sequence. Finally, the GTA solution was subjected to rotary evaporation and stored in a vacuum desiccator for further use;
[0037] 2) Add the polypeptide ε-polylysine to 45-55°C deionized water to dissolve and stir. After 30 minutes, add GTA modified small molecules and keep stirring for 40-60 hours, wherein the polypeptide ε-polylysine and The molar ratio of GTA is (3-5):1. Finally, the resulting GEPL polymer was purified using dialysis tubing (mwco1000) f...
Embodiment 1
[0046] 1) The preparation method of GTA-modified small molecules: firstly add 10mmol glycerol into 40mL anhydrous chloroform, stir and dissolve at room temperature, and then add 45mmol triethylamine. Then 45 mmol of acryloyl chloride was added dropwise to the mixture under ice-cooling conditions. The reaction temperature was increased to 50°C and stirred for 48 hours. After the reaction is complete, use HCl, NaHCO 3 、H 2 O washes the mixture. Finally, the GTA solution was subjected to rotary evaporation and stored in a vacuum desiccator for further use;
[0047] 2) Preparation method of GEPL polymer: add 10mmolε-polylysine to 50°C deionized water to dissolve and stir, after 30 minutes, add 3mmol GTA modified small molecule and keep stirring for 48 hours. Finally, the resulting GEPL polymer was purified using dialysis tubing (mwco1000) for 3 days. After lyophilization, the final GEPL polymers were collected and stored for further use;
[0048] 3) Gene-activated antibacter...
Embodiment 2
[0057] 1) The preparation method of GTA-modified small molecules: 10 mmol of glycerin was first added into 35 mL of anhydrous chloroform, stirred and dissolved at room temperature, and then 50 mmol of triethylamine was added. Then 45 mmol of acryloyl chloride was added dropwise to the mixture under ice-cooling conditions. The reaction temperature was increased to 45°C and stirred for 60 hours. After the reaction is complete, use HCl, NaHCO 3 、H 2 O washes the mixture. Finally, the GTA solution was subjected to rotary evaporation and stored in a vacuum desiccator for further use;
[0058] 2) Preparation method of GEPL polymer: add 10mmolε-polylysine to 50°C deionized water for dissolving and stirring, after 30 minutes, add 2.8mmol GTA modified small molecule and keep stirring for 48 hours. Finally, the resulting GEPL polymer was purified using dialysis tubing (mwco1000) for 3 days. After lyophilization, the final GEPL polymers were collected and stored for further use;
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