Application of antibacterial peptide compound DM80Bu20 in preparation of antibacterial toothpaste
A technology of dm80bu20 and dm80bu200.001 is applied in the field of oral care to achieve the effects of long-term storage, reducing gum bleeding and reducing dental calculus
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Embodiment 1
[0042] Example 1 DM80Bu20 in vitro antibacterial activity test
[0043] 1. Experimental method
[0044] (1) Strain culture
[0045] Candida albicans ATCC SC5314 was cultured in RPMI 1640 medium at 37°C with 80% humidity and 5% CO 2 Incubation under the conditions;
[0046] Staphylococcus aureus ATCC 6538 in TSB medium at 37°C, 5% CO 2 Incubation under the conditions;
[0047] Methicillin-resistant Staphylococcusaureus 18908 in TSB medium, 37°C, 5% CO 2 Incubation under the conditions;
[0048] Porphyromonas gingivalis ATCC 33277 was cultured in BHI medium with hemin (haemin, 5 μg / mL) and vitamin K (menadione, 1 μg / mL) at 37°C under strict anaerobic conditions;
[0049] Streptococcus mutans Clarke Streptococcus mutansClarkeStreptococcus mutans ATCC UA159 in medium Brain Heart Infusion Agar / Broth, 37°C, 5% CO 2 cultivated under conditions;
[0050] Streptococcus sanguis ATCC 10556 in medium Brain HeartInfusionAgar / Broth, 37°C, 5% CO 2 cultivated under conditions.
[00...
Embodiment 2
[0063] Embodiment 2 compound DM80Bu20 hemolytic activity test
[0064] 1. Experimental method
[0065] Fresh human blood was washed three times with Tris buffer (TBS), and the collected human red blood cells (hRBCs) were diluted to 5% (v / v) with TBS. On a 96-well plate, compound DM80Bu20 was diluted to 3.13-400 μg / mL by two-fold serial dilution. After mixing equal volumes of hRBCs suspension and compound DM80Bu20 solution, they were incubated at 37°C for 1 hour. TBS was used as the blank control, and Triton X-100 (TBS) 3.2 μg / ml and hRBCs equal volume mixed solution were used as the positive control. After centrifugation, transfer 80 μl of the supernatant in each well to another 96-well plate, read the OD value of the solution to be tested in the 96-well plate at 405 nm with a microplate reader, and calculate the hemolysis percentage by the following formula:
[0066]
[0067] 2. Experimental results
[0068] Hemolytic activity of DM80Bu20 (HC 50 ) was 200 μg / mL, indic...
Embodiment 3
[0069] Cytotoxic activity test of embodiment 3 compound DM80Bu20
[0070] 1. Experimental method
[0071] Prepare a single cell suspension with DMEM medium containing 10% fetal bovine serum, and the cell density is 1×10 6 CFU / mL, and 1×10 per well 4 Cells were seeded into a 96-well plate with a volume of 100 μL per well. Cells were incubated at 37°C for 24 hours. After the old medium was removed, medium containing different concentrations of compound DM80Bu20 was added, and three replicate wells were set for each concentration. After the cells were cultured at 37° C. for 24 hours, 10 μL of MTT solution (5 mg / mL, prepared in PBS) was added to each well, and the incubation was continued for 4 hours to terminate the culture. Carefully suck off the culture supernatant in the wells, add 150 μL DMSO to each well, and shake on the shaker for 10 minutes to fully dissolve the crystals. On the same 96-well plate, cells treated without any compound DM80Bu20 were included as a contro...
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