| Summary: | 碩士 === 義守大學 === 生物醫學工程學系 === 101 === Curcumin is the main component of traditional medicine turmeric, an orange powder extracted from plant. Curcumin is able to directly capture free radicals, inhibit the formation of free radicals, and promote apoptosis effect. In recent years, studies have shown that curcumin was able to inhibit colon cancer, prostate cancer, lung cancer and induce apoptosis in various tumor cells. However, curcumin is insoluble in water, low bioavailability, short half-life and thereby limits its therapeutic application. It is well known that natural polymer has excellent biocompatibility, biodegradability and is widely used in biomedical materials and clinical treatment. Particles have good surface area/volume ratio, diffusion resistance, as well as unrestricted format of application. In this study, natural polymer hyaluronic acid, gelatin, chitosan were used in the preparation of nano encapsulation of curcumin and to study the combination effects of nanoparticles and curcumin on human lung adenocarcinoma A549 cells could reduce the amount of drug, duration of treatment and promote the apoptotic effect of curcumin. The parallel electric field was used to prepare hyaluronic acid, gelatin, chitosan nanoparticles and their particle size were 2.77 ± 0.89 nm, 3.21 ± 1.19 nm, and 4.02 ± 0.70 nm, respectively. But the particle size of nanoparticles after encapsulating curcumin(2.5 μg / mL)as 24.34 ± 4.24 nm, 26.97 ± 4.77 nm, 29.18 ± 4.28 nm, the particle size of nanoparticles after encapsulating curcumin(5.0 μg / mL)as 31.23 ± 4.69 nm, 28.17 ± 4.16 nm, 54.66 ± 4.67 nm.
The encapsulation efficiency of curcumin(2.5 μg / mL) in hyaluronic acid was 63.13 ± 3.18 (%); curcumin in gelatin was 73.64 ± 2.53 (%);curcumin in chitosan was 45.19 ± 6.28 (%);The encapsulation efficiency of curcumin(5.0 μg / mL) in hyaluronic acid was 62.63 ± 1.74 (%); curcumin in gelatin was 49.47 ± 0.97 (%); curcumin in chitosan was 81.26 ± 2.22 (%). MTT assay showed that nanoparticles with curcumin have low cells viability after 24 hours of treatment and even lower after 48 hours of treatment, when compared to control group, indicating that it has the effect of inhibiting proliferation or cytotoxicity on A549 cells. To confirm the result of apoptosis, flow cytometry was performed and the results showed that the percentage of Annexin V-FITC of gelatin-coated curcumin group, hyaluronic acid coated curcumin group, chitosan-coated curcumin group were 42.3%, 31.0%, 25.1% after 48 hours of treatment, compared to the simple drug curcumin group which has only 12.5% in early apoptosis. The result of JC-1 after 48 hours of treatment showed similar outcome. Knowing that curcumin concentration at 2.5 μg / mL would have apoptotic effect require 48 hours of treatment, a higher concentration of curcumin (5.0 μg / mL) was tested. The results of Annexin V after 6 hours of treatment showed that gelatin-coated curcumin group, hyaluronic acid-coated curcumin group, chitosan-coated curcumin group has apoptotic rates of 41.7%, 30.9%, and 44.7%, respectively, whereas curcumin alone group has apoptosis rate of 18.1% of apoptosis. The results of JC-1 showed similar outcome. In addition, the endocytosis of nanoparticles into cells was observed by mixing Did oil with gelatin or hyaluronic acid, producing nanoparticles and culture into A549 cells. Red fluorescence of Did oil with gelatin nanoparticles can be observed after 10 minutes culture, 2 hours culture for Did oil with chitosan nanoparticles; indicating that different materials affect the endocytosis of nanoparticles into A549 cells.
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