A preparation method of hyaluronic acid-melanin nanoparticles
A technology of hyaluronic acid and nanoparticles, which is applied in the field of preparation of hyaluronic acid-melanin nanoparticles, can solve the problems of long-term toxicity, toxic reactions, and poor biological metabolism, and achieve the goal of improving cell uptake efficiency and excellent biocompatibility Effect
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Embodiment 1
[0040] This embodiment provides a method for preparing hyaluronic acid-melanin nanoparticles, which specifically includes the following preparation steps:
[0041] Step 1: Separation and extraction of natural melanin nanoparticles
[0042] Thaw the frozen North Pacific squid (Ommastrephes bartrami) ink sac, squeeze out the epidermal membrane and internal reticular membrane, filter the squid ink, soak it in an equal volume of ice water overnight, discard the supernatant by centrifugation, and add 2 to 3 times the volume of the precipitated part Then centrifuge again, repeating 4-6 times, until the color of the supernatant after centrifugation is basically no black; the obtained precipitate is vacuum freeze-dried, and the melanin nanoparticle sample is obtained;
[0043] Step 2: Amination of Hyaluronic Acid (HA)
[0044] Hyaluronic acid (100mg, 0.1mmol) with a molecular weight of 6KDa was dissolved in 25ml of PBS buffer solution (0.01M, pH=7.4) to make a 4mg / mL solution. After st...
Embodiment 2
[0048] This embodiment provides a method for preparing hyaluronic acid-melanin nanoparticles. The difference between this embodiment and Example 1 is that in step 3, the melanin nanoparticles prepared in step 1 are weighed and dissolved in Tris- A melanin nanoparticle solution with a concentration of 1 mg / mL was made in HCL buffer solution; 12 mg of aminated hyaluronic acid prepared in step 2 was weighed and dissolved in 8 mL of Tris-HCL buffer solution, and 2 mL of 1 mg / mL was added thereto The melanin nanoparticle solution was stirred at room temperature in the dark for 24 hours; after the reaction, the reaction solution was dialyzed in deionized water with a (MWCO 3500) dialysis bag for 48 hours, and the reaction solution was lyophilized and stored at 4°C for use.
Embodiment 3
[0050] This embodiment provides a method for preparing hyaluronic acid-melanin nanoparticles. The difference between this embodiment and Example 1 is that in step 3, the melanin nanoparticles prepared in step 1 are weighed and dissolved in Tris- A solution of melanin nanoparticles with a concentration of 1 mg / mL was prepared in HCL buffer solution; 10 mg of hyaluronic acid with an amination ratio of 40% was weighed and dissolved in 8 mL of Tris-HCL buffer solution, and 2 mL of 1 mg / mL melanin nanoparticles were added to it. The particle solution was stirred at room temperature in the dark for 24 hours; after the reaction, the reaction solution was dialyzed in deionized water for 48 hours with a dialysis bag with a molecular weight of 3500 (MWCO 3500), and the reaction solution was lyophilized and stored at 4°C for use.
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