Preparation method and application of chemotherapy-phototherapy synergetic anti-tumor microsphere
An anti-tumor and microsphere technology, used in anti-tumor drugs, photodynamic therapy, medical preparations with inactive ingredients, etc., can solve the problems of low feasibility, excessive use, difficult industrial production, etc., and achieve strong compliance , enhance drug resistance, and have the effect of industrial production feasibility
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Embodiment 1
[0031] Take two 50mL dry screw-top bottles and add the prepared 2% sodium alginate solution, and then add 40 μg / mL chemotherapeutic drug doxorubicin (DOX) and 10% cellulose nanocrystals (CNC) to one of the bottles, Add 2.5 μg / mL photosensitizer indocyanine green (ICG) to the other bottle, and mix the two bottles of solutions evenly, as the internal and external phases of chemotherapy-phototherapy synergistic anti-tumor core-shell microspheres. Take another receiving dish without a cover, add an aqueous solution containing 2% calcium chloride and 0.15% chitosan, and the solution depth exceeds 1 cm, as a receiving bath for the microspheres. Using gas-assisted method, add sodium alginate solution containing DOX and CNC to the internal phase channel, add sodium alginate solution containing ICG to the external phase channel, and then pass nitrogen gas into the outermost layer at a flow rate of 6L / min to form core-shell droplets, and solidified by a receiving bath to finally form an...
Embodiment 2
[0033] Both the free ICG and the microspheres were placed in a cuvette, and the full-wavelength absorbance was measured with a UV-visible spectrophotometer, as shown in Figure 4 As shown in (A), both the free ICG and the ICG encapsulated in microspheres have the maximum absorption peak at 780nm.
[0034] Both free ICG and microspheres were incubated in PBS at 37°C for 5 days. The absorbance at 780 nm was measured with a UV-vis spectrophotometer every day, such as Figure 4 As shown in (B), the free ICG was obviously degraded, but its stability was significantly improved after being encapsulated by microspheres.
[0035] Both free ICG and microspheres were placed in 25 μg / mL DPBF solution, and the DPBF solution without any other substances was used as a blank control. Use 0.8W / cm 2 808nm near-infrared laser irradiation for 5 minutes, and measure the absorbance at 410nm with a UV-visible spectrophotometer every minute, as Figure 4 (C) shown. Both the free ICG and the ICG ...
Embodiment 3
[0039] HepG-2 cells were placed in DMEM medium containing 10% fetal bovine serum and cultured at 37°C in a cell culture incubator with 95% sterile air and 5% carbon dioxide environment. Change the cell culture medium every two days.
[0040] 2',7'-dichlorodihydrofluorescein diacetate (DCFH-DA) was used as a fluorescent probe to measure the level of intracellular ROS. HepG-2 cells were seeded in 12-well plates at a cell density of 1×10 5 One per well, cultured for 24 hours, then replaced the previous medium with free ICG or microspheres and cultured for 8 hours, and then added 10 μg / mL DCFH-DA. After incubating for 20 minutes, remove the medium, wash the cells three times with PBS buffer, and use 808nm laser (0.8W / cm2 ) for 3 minutes, observed cell morphology, and imaged with a fluorescent microscope (IX 53, OLYMPUS, Japan), such as Figure 7 shown.
[0041] The killing effect of chemotherapy-phototherapy combined with anti-tumor core-shell microspheres on cancer cell HepG-2...
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