Composite hydrogel based on zwitterions and keratin and preparation method thereof
A composite hydrogel and amphoteric ion technology, applied in medical science, bandages, etc., can solve the problems of limited development and application, difficult dressing, surface separation of newborn tissues, etc.
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Embodiment 1
[0023] 1. Synthesis of the crosslinker BACy
[0024] The cross-linking agent BACy is synthesized by acrylated two amino groups of cystamine dihydrochloride, and the synthetic route is as follows:
[0025]
[0026] Cystamine hydrochloride (5.8 g, 0.025 mol) was added to a 100 mL three-necked flask equipped with a thermometer and two 10 mL constant pressure dropping funnels, and 25 mL of deionized water was added. After the mixture was cooled with ice water, a solution of acryloyl chloride (4.65 g, 0.05 mol) in dichloromethane (10 mL) and NaOH solution (4.0 g, 0.1 mol; 10 mL) were slowly added dropwise under stirring, and the dropping time was controlled at 0.5 More than h, the temperature is always kept at 0-5°C during the dropping process. After the dropwise addition was complete, the reaction mixture was stirred at room temperature for more than 2 h. The organic phase was separated and extracted several times with dichloromethane (5 x 50 mL). The collected organic phase...
Embodiment 2
[0034] Anti-protein adsorption performance test:
[0035] Fluorescein isothiocyanate (FITC)-labeled bovine serum albumin (BSA) was prepared by conjugating FITC to BSA to prepare FITC-BSA. First with 100mM NaHCO 3 (pH=9) was used as a solvent to prepare a 10 mg / mL BSA solution, and FITC was dissolved in DMSO to prepare a 1 mg / mL solution for later use. FITC was slowly added dropwise to BSA under the protection from light, and reacted for 2h. After the end, with 100mM NaHCO 3 (pH=9) Dialyze in the dark until the dialysate is colorless, and then dialyze three times with deionized water to obtain a FITC-BSA solution for later use.
[0036] The flaky PDMAPS and PDMAPS / Keratin hydrogel with a cross-linking degree of 5% were placed in a 6-well cell culture plate, and an equal amount of FITC-BSA solution was added to the sample wells to completely immerse the gel. Incubate at 4°C in the dark for 12 hours, discard the FITC-BSA solution, wash with PBS buffer (pH=7.4) three times, an...
Embodiment 3
[0039] Hydrogel interaction with L929 cells:
[0040]Before cell culture, the prepared hydrogel was detoxified in the specified medium for 5 days. During this process, the unreacted cross-linker was effectively removed from the hydrogel, and the essential nutrients in the hydrogel were enriched. In order to study the interaction between hydrogel and cells, the round hydrogel was sterilized by ultraviolet irradiation in a clean bench and placed in a 24-well cell culture plate. Digest L-929 cells with trypsin and make 3×10 4 cells / mL of cell suspension. Add 1 mL of cell suspension to each well, 37°C, 5% CO 2 Cultured in the incubator for 3d. After the end, add 100 μL MTT solution with a concentration of 0.5 mg / mL to each well, continue to cultivate in the incubator for 4 hours, discard the MTT solution and cell culture medium, add 500 μL DMSO after washing with PBS solution, shake for 30 minutes in the dark, and transfer the solution to 96 In the well plate, use a microplat...
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