Chitosan Inhibits Helicobacter pylori Growth and Urease Production and Prevents Its Infection of Human Gastric Carcinoma Cells
Abstract
:1. Introduction
2. Results and Discussion
2.1. Effects of DD95 Chitosan on Growth and Urease Production of H. pylori
2.2. Combination of DD95 Chitosan and Antibiotics against H. pylori
2.3. Adhesion of H. pylori to Human Gastric Carcinoma Cells (TSGH9201) in the Presence of DD95 Chitosan
3. Material and Methods
3.1. Bacterial Strains, Cell Line, and Chemicals
3.2. DD95 Chitosan Preparation
3.3. Culture Conditions
3.4. Antibacterial Test
3.5. Combination Effects of Antibiotic and DD95 Chitosan against H. pylori
3.6. Urease Activity Assay
3.7. Adherence of Fluorescein-Labeled H. pylori to Human Gastric Carcinoma Cells in the Presence of DD95 Chitosan
3.8. Statistical Analysis
4. Conclusions
Author Contributions
Funding
Conflicts of Interest
References
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pH | BCRC 10726 | No.123 | No.125 | ||||||
---|---|---|---|---|---|---|---|---|---|
Bacterial Count 2 (log CFU/mL) | Survival (%) | Bacterial Count (log CFU/mL) | Survival (%) | Bacterial Count (log CFU/mL) | Survival (%) | ||||
Control | Chitosan | Control | Chitosan | Control | Chitosan | ||||
2 | 5.49 ± 0.01 D | 3.22 ± 0.06 | 0.53 D,3 | 5.41 ± 0.02 D | 3.23 ± 0.03 | 0.65 D | 5.20 ± 0.03 E | 3.27 ± 0.03 | 1.17 D |
3 | 6.62 ± 0.01 C | 4.47 ± 0.01 | 0.70 D | 6.66 ± 0.05 C | 4.95 ± 0.06 | 1.94 D | 6.59 ± 0.02 D | 4.99 ± 0.05 | 3.51 D |
4 | 6.83 ± 0.03 B | 5.50 ± 0.00 | 4.71 C | 6.85 ± 0.05 B | 5.36 ± 0.05 | 3.22 D | 6.79 ± 0.05 C | 5.44 ± 0.03 | 4.51 D |
5 | 6.92 ± 0.03 B | 5.76 ± 0.01 | 6.93 B | 6.90 ± 0.06 B | 5.77 ± 0.03 | 7.46 C | 6.92 ± 0.04 B | 5.85 ± 0.03 | 8.62 C |
6 | 6.97 ± 0.02 A | 5.89 ± 0.00 | 8.29 B | 7.10 ± 0.07 A | 6.04 ± 0.08 | 8.74 C | 7.09 ± 0.02 A | 6.09 ± 0.03 | 10.02 C |
7 | 6.97 ± 0.01 A | 6.93 ± 0.02 | 89.90 A | 7.06 ± 0.06 A | 7.02 ± 0.06 | 90.72 B | 7.06 ± 0.02 A | 7.02 ± 0.04 | 89.89 B |
8 | 6.97 ± 0.00 A | 6.93 ± 0.01 | 91.63 A | 7.03 ± 0.01 A | 7.02 ± 0.01 | 96.63 A | 7.06 ± 0.02 A | 7.04 ± 0.03 | 94.30 A |
pH | BCRC 10726 | No. 123 | No. 125 | ||||||
---|---|---|---|---|---|---|---|---|---|
OD550 | Relative Urease Activity (%) | OD550 | Relative Urease Activity (%) | OD550 | Relative Urease Activity (%) | ||||
Control | Chitosan | Control | Chitosan | Control | Chitosan | ||||
2 | 0.40 ± 0.00 C | 0.15 ± 0.00 2 | 37.84 ± 0.35 D,3 | 0.39 ± 0.01 C | 0.18 ± 0.00 | 45.58 ± 1.52 F | 0.41 ± 0.00 C | 0.19 ± 0.01 | 46.58 ± 0.37 F |
3 | 0.41 ± 0.00 C | 0.18 ± 0.01 | 43.72 ± 0.35 C | 0.40 ± 0.01 C | 0.19 ± 0.01 | 48.63 ± 1.93 E | 0.42 ± 0.00 C | 0.22 ± 0.00 | 51.93 ± 0.64 E |
4 | 0.44 ± 0.01 A | 0.21 ± 0.00 | 47.82 ± 0.98 B | 0.44 ± 0.01 A | 0.22 ± 0.00 | 49.25 ± 0.58 D,E | 0.45 ± 0.00 A | 0.25 ± 0.01 | 54.84 ± 0.54 D |
5 | 0.42 ± 0.00 B | 0.20 ± 0.01 | 48.61 ± 1.08 B | 0.43 ± 0.01 A,B | 0.22 ± 0.00 | 50.51 ± 0.69 C,D | 0.44 ± 0.00 B | 0.24 ± 0.00 | 56.02 ± 0.41 C,D |
6 | 0.41 ± 0.00 C | 0.20 ± 0.01 | 49.51 ± 0.25 B | 0.42 ± 0.00 B | 0.22 ± 0.01 | 51.34 ± 0.51 C | 0.43 ± 0.01 B | 0.25 ± 0.00 | 47.39 ± 1.09 C |
7 | 0.20 ± 0.00 D | 0.20 ± 0.00 | 97.54 ± 0.50 A | 0.21 ± 0.01 D | 0.20 ± 0.01 | 93.45 ± 1.06 B | 0.21 ± 0.01 D | 0.20 ± 0.01 | 94.85 ± 0.61 B |
8 | 0.20 ± 0.01 D | 0.20 ± 0.01 | 97.35 ± 0.71 A | 0.21 ± 0.01 D | 0.20 ± 0.01 | 97.89 ± 0.24 A | 0.21 ± 0.01 D | 0.20 ± 0.01 | 97.89 ± 0.63 A |
Antibiotic (μg/mL) | BCRC 10726 | No. 123 | No. 125 | ||||
---|---|---|---|---|---|---|---|
Bacterial Count (Log CFU/mL) | Bacterial Count (Log CFU/mL) | Bacterial Count (Log CFU/mL) | |||||
Chitosan conc. (μg/mL) | Chitosan conc. (μg/mL) | Chitosan conc. (μg/mL) | |||||
0 | 4000 | 0 | 4000 | 0 | 4000 | ||
Control | 7.20 ± 0.03 1 | 5.32 ± 0.10 | 7.61 ± 0.02 | 5.13 ±0.05 | 7.59 ± 0.01 | 5.39 ± 0.07 | |
Amoxicillin | 50 | 4.36 ± 0.01 | 0.00 ± 0.00 | 4.42 ± 0.01 | 0.00 ± 0.00 | 4.56 ± 0.01 | 0.00 ± 0.00 |
100 | 3.26 ± 0.01 | 0.00 ± 0.00 | 4.31 ± 0.01 | 0.00 ± 0.00 | 4.33 ± 0.01 | 0.00 ± 0.00 | |
200 | 2.25 ± 0.05 | 0.00 ± 0.00 | 3.74 ± 0.04 | 0.00 ± 0.00 | 4.04 ± 0.02 | 0.00 ± 0.00 | |
4000 | 0.00 ± 0.00 | - | 0.00 ± 0.00 | - | 0.00 ± 0.00 | - | |
Tetracycline | 50 | 5.69 ± 0.02 | 3.24 ± 0.02 | 5.37 ± 0.05 | 3.84 ± 0.03 | 5.39 ± 0.01 | 3.92 ± 0.02 |
100 | 5.02 ± 0.03 | 0.00 ± 0.00 | 4.75 ± 0.01 | 0.00 ± 0.00 | 4.78 ± 0.01 | 0.00 ± 0.00 | |
200 | 3.98 ± 0.02 | 0.00 ± 0.00 | 2.80 ± 0.04 | 0.00 ± 0.00 | 3.22 ± 0.01 | 0.00 ± 0.00 | |
4000 | 0.00 ± 0.00 | - | 0.00 ± 0.00 | - | 0.00 ± 0.00 | - | |
Metronidazole | 50 | 4.69 ± 0.01 | 2.46 ± 0.02 | 5.07 ± 0.01 | 3.38 ± 0.02 | 5.11 ± 0.02 | 3.64 ± 0.02 |
100 | 4.55 ± 0.02 | 0.00 ± 0.00 | 3.89 ± 0.00 | 0.00 ± 0.00 | 3.95 ± 0.00 | 0.00 ± 0.00 | |
200 | 4.28 ± 0.02 | 0.00 ± 0.00 | 3.10 ± 0.02 | 0.00 ± 0.00 | 3.34 ± 0.02 | 0.00 ± 0.00 | |
4000 | 0.00 ± 0.00 | - | 0.00 ± 0.00 | - | 0.00 ± 0.00 | - |
Antibiotic (μg/mL) | BCRC 10726 | No. 123 | No. 125 | ||||
---|---|---|---|---|---|---|---|
Bacterial Count (Log CFU/mL) | Bacterial Count (Log CFU/mL) | Bacterial Count (Log CFU/mL) | |||||
Chitosan (μg/mL) | Chitosan (μg/mL) | Chitosan (μg/mL) | |||||
0 | 4000 | 0 | 4000 | 0 | 4000 | ||
Control | 6.70 ± 0.03 1 | 3.97 ± 0.06 | 6.61 ± 0.02 | 4.28 ± 0.02 | 6.66 ± 0.04 | 4.52 ± 0.01 | |
Amoxicillin | 50 | 4.08 ± 0.03 | 0.00 ± 0.00 | 3.83 ± 0.01 | 0.00 ± 0.00 | 3.84 ± 0.00 | 0.00 ± 0.00 |
100 | 3.44 ± 0.02 | 0.00 ± 0.00 | 3.60 ± 0.01 | 0.00 ± 0.00 | 3.63 ± 0.01 | 0.00 ± 0.00 | |
200 | 3.08 ± 0.03 | 0.00 ± 0.00 | 3.30 ± 0.01 | 0.00 ± 0.00 | 3.40 ± 0.00 | 0.00 ± 0.00 | |
4000 | 0.00 ± 0.00 | - | 0.00 ± 0.00 | - | 0.00 ± 0.00 | - | |
Tetracycline | 50 | 5.23 ± 0.03 | 0.00 ± 0.00 | 4.39 ± 0.02 | 0.00 ± 0.00 | 4.18 ± 0.02 | 0.00 ± 0.00 |
100 | 4.58 ± 0.01 | 0.00 ± 0.00 | 3.75 ± 0.01 | 0.00 ± 0.00 | 3.02 ± 0.02 | 0.00 ± 0.00 | |
200 | 3.39 ± 0.01 | 0.00 ± 0.00 | 1.79 ± 0.01 | 0.00 ± 0.00 | 2.47 ± 0.02 | 0.00 ± 0.00 | |
4000 | 0.00 ± 0.00 | - | 0.00 ± 0.00 | - | 0.00 ± 0.00 | - | |
Metronidazole | 50 | 4.23 ± 0.00 | 0.00 ± 0.00 | 4.08 ± 0.01 | 0.00 ± 0.00 | 4.46 ± 0.00 | 0.00 ± 0.00 |
100 | 4.10 ± 0.05 | 0.00 ± 0.00 | 2.91 ± 0.00 | 0.00 ± 0.00 | 3.87 ± 0.03 | 0.00 ± 0.00 | |
200 | 3.84 ± 0.02 | 0.00 ± 0.00 | 2.10 ± 0.02 | 0.00 ± 0.00 | 2.49 ± 0.02 | 0.00 ± 0.00 | |
4000 | 0.00 ± 0.00 | - | 0.00 ± 0.00 | - | 0.00 ± 0.00 | - |
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Chang, S.-H.; Hsieh, P.-L.; Tsai, G.-J. Chitosan Inhibits Helicobacter pylori Growth and Urease Production and Prevents Its Infection of Human Gastric Carcinoma Cells. Mar. Drugs 2020, 18, 542. https://doi.org/10.3390/md18110542
Chang S-H, Hsieh P-L, Tsai G-J. Chitosan Inhibits Helicobacter pylori Growth and Urease Production and Prevents Its Infection of Human Gastric Carcinoma Cells. Marine Drugs. 2020; 18(11):542. https://doi.org/10.3390/md18110542
Chicago/Turabian StyleChang, Shun-Hsien, Pei-Ling Hsieh, and Guo-Jane Tsai. 2020. "Chitosan Inhibits Helicobacter pylori Growth and Urease Production and Prevents Its Infection of Human Gastric Carcinoma Cells" Marine Drugs 18, no. 11: 542. https://doi.org/10.3390/md18110542