Hyaluronic acid (HA)-polyethyleneimine (PEI) bonded copolymer as well as preparation method and application thereof
A technology of polyethyleneimine and hyaluronic acid, which is applied in the field of biomedical materials, can solve problems such as limited application, and achieve the effects of prolonging the action time, improving safety, and enhancing adhesion
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Embodiment 1b
[0053] The synthesis of embodiment 1bPEI-HA complex
[0054] a. Dissolve 10 mL of HA with a concentration of 2 mg / mL in 10 mL of bPEI with a concentration of 40 mg / mL, and adjust the pH value of the reactant to 6.5 with 1.0 mol / L HCl solution.
[0055] b. Dissolve 40.4 mg EDC·HCl and 28.5 mg HOBt in 1 ml of 50% by volume DMSO solution.
[0056] c. Add the solution obtained in step b to the solution obtained in step a. After stirring and mixing at room temperature for 24 hours, adjust the pH to 7.0 with NaOH with a concentration of 1.0 mol / L.
[0057] d. Pour the solution obtained in step c into a cleaned 10kDa dialysis bag, dialyze in 100mmol / L NaCl solution for 2 days, dialyze in 25wt% ethanol for one day, and dialyze in purified water for one day to obtain 1 μg / μL of bPEI-HA complex The solution was freeze-dried to obtain a white powdery solid. reuse 1 H-NMR analysis detected the degree of bPEI modification of HA.
[0058] use 1 H-NMR analysis to detect the degree of bP...
Embodiment 2b
[0059] The effect detection experiment of embodiment 2bPEI-HA complex
[0060] (1) Preparation of siRNA / bPEI-HA complex
[0061] Take the anti-PGL3-Luc siRNA with a concentration of 0.75 μg / μL and the bPEI-HA complex with a concentration of 0.75 μg / μL and mix them in volume ratios of 1:1, 1:3 and 1:5, respectively. As shown in Table 1, incubate at room temperature for 15 minutes to make siRNA / bPEI-HA complex. Finally, 3 μL of 5 mol / L NaCl solution was added to make the final concentration of siRNA / bPEI-HA complex reach 150 mmol / L.
[0062] Mix 0.75 μg / μL anti-PGL3-Luc siRNA with 0.75 μg / μL bPEI at a volume ratio of 1:1, 1:3, and 1:5 respectively. The mixing method is shown in Table 2, and incubated at room temperature 15 minutes to make siRNA / bPEI complexes. Finally, 3 μL of 5 mol / L NaCl solution was added to make the final concentration of the siRNA / bPEI complex reach 150 mmol / L.
[0063] Table 1. Mixing ratio of siRNA and bPEI-HA complex
[0064]
[0065] Table 2. Mi...
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