Cyclic gamma-polyglutamic acid modified hydrogel loaded nano sponge detoxification system and preparation method thereof
A polyglutamic acid and nano-sponge technology, applied in medical science, bandages, etc., can solve problems such as unreported, and achieve the effect of good biocompatibility
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Embodiment 1
[0046] Embodiment 1: the preparation of NS
[0047] ①Preparation of PLGA nano-core: 0.67dl / g carboxyl-containing 50:50 PLGA monomer polymer was prepared into 60nm PLGA nanoparticles by nano-precipitation method. First, dissolve PLGA monomer in acetone to prepare PLGA acetone solution (5mg / ml); then, add 1ml PLGA acetone solution to 3ml deionized water, stir at room temperature for 2h; finally, centrifuge with AmiconUltra-4 with a molecular weight cut-off of 10kDa The filter was filtered to obtain PLGA nanoparticles ( figure 1 ). In addition, to prepare fluorescently labeled NSs for subsequent tracking, 0.1 wt% Did fluorescent dye was added to PLGA acetone solution before polymerization, and Did-labeled PLGA nanoparticles were similarly prepared ( figure 1 ). Observed under the transmission electron microscope, it can be seen that the typical nano-scale particle structure ( figure 2 ). The hydrated particle size and zeta potential ( image 3 , 4).
[0048] ②Preparation ...
Embodiment 2
[0050] Example 2: Preparation of cyclo-γ-PGA modified PNIPAM hydrogel loaded NS detoxification system
[0051]①cyclo-γ-PGA modified PNIPAM hydrogel: mix the two substances in sequence according to PNIPAM / cyclo-γ-PGA ratio of 100:0, 75:25, 50:50, and 25:75, and the total concentration of the solution is 20% ( w / v), mix well at room temperature for 3 hours, remove air bubbles overnight at 4°C, and gel in a water bath at 37°C. The dehydration phenomenon of PNIPAM hydrogel can be observed when PNIPAM / cyclo-γ-PGA is 75:25 or 50:50. effectively improved ( Figure 7 ,8). Under the scanning electron microscope, it was observed that when the ratio of PNIPAM / cyclo-γ-PGA was 75:25, the hydrogel showed a typical porous structure ( Figure 9 ).
[0052] ②Mix the final concentration of 2mg / ml NS, 150mg / mL PNIPAM and 50mg / mLcyclo-γ-PGA, mix thoroughly at room temperature for 3 hours, remove air bubbles overnight at 4°C, and gel in a water bath at 37°C. The structure of hydrogel-loaded NS...
Embodiment 3
[0053] Example 3: Biocompatibility of cyclo-γ-PGA modified PNIPAM hydrogel loaded NS detoxification system
[0054] To detect the cytotoxic effect of cyclo-γ-PGA modified PNIPAM hydrogel loading NS detoxification system. First, prepare cyclo-γ-PGA modified PNIPAM hydrogel-loaded NS detoxification system extract, immerse the prepared detoxification system in serum-free DMEM medium at a ratio of 0.1 g / mL, and extract at 37°C for 24 hours , collect the culture medium, and pass through a 0.22 μm filter membrane to sterilize for later use. After cell counting, L929 cells were divided into 1 × 10 4 Inoculate each well into a 96-well plate, cultivate overnight until it adheres to the wall, suck away the medium, then dilute the extraction solution according to the concentration gradient of 100%, 50%, and 25%, add 100 μL of different concentration extraction solutions to each well, and cultivate for 24 hours or 48h, after taking it out, add 10μL CCK-8 reagent to each well, incubate f...
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