Study on Optimizing Novel Antimicrobial Peptides with Bifunctional Activity to Prevent and Treat Peri-Implant Disease
Abstract
:1. Introduction
2. Results
2.1. Properties of Antimicrobial Peptides
2.2. Antimicrobial Activity against S. gordonii and F. nucleatum
2.3. Biofilm Inhibition
2.4. Scanning Electron Microscope (SEM)
2.5. Biocompatibility of Antimicrobial Peptides
2.6. Effect of Peptides on the Expression of Pro-Inflammatory and Anti-Inflammatory Genes of RAW 264.7
2.7. Effects of Antimicrobial Peptides on Polarized Morphology of RAW264.7
2.8. Antimicrobial Peptides Pair Regulation of NF-κB-p65 Signal Pathway during RAW 264.7 Polarization
2.9. Regulation of Conditioned Medium on Osteogenic Ability of Osteoblasts MC3T3-E1
3. Discussion
4. Materials and Methods
4.1. Synthesis and Characterization of Antimicrobial Peptides
4.2. Behaviors of S. gordonii and F. nucleatum Cultured with the Different Antimicrobial Peptides
4.2.1. Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC)
4.2.2. Biofilm Susceptibility Assay
4.2.3. Confocal Laser Scanning Microscopy (CLSM)
4.2.4. Scanning Electron Microscopy (SEM)
4.3. Behaviors of Macrophages RAW 264.7 Cultured with the Different Antimicrobial Peptides
4.3.1. Cell Culture
4.3.2. Cell Proliferation Assay
4.3.3. Gene Expression of Cell Polarization
4.3.4. Cell Morphology and p65 Immunofluorescence Staining
4.3.5. Flow Cytometry
4.4. Behavior of MC3T3-E1 Cells in Different Antimicrobial Peptides Conditioned Media (CM)
4.4.1. Preparation of Conditioned Medium
4.4.2. Cell Culture
4.4.3. Osteogenic-Related Gene Expression
4.4.4. Alkaline Phosphatase Staining Assay
4.4.5. Alizarin Red S (ARS) Staining
4.4.6. Western Blot Analysis
4.5. Statistical Analysis
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Bacteria | MIC (μg/mL) | MBC (μg/mL) | ||
---|---|---|---|---|
KR−1 | KR−2 | KR−1 | KR−2 | |
S. gordonii | 125 | 50 | 250 | 150 |
F. nucleatum | 100 | 50 | 200 | 100 |
Gene (Mouse) | Forward Primer Sequence (5′-3′) | Reverse Primer Sequence (5′-3′) |
---|---|---|
CD80 | CCTCAAGTTTCCATGTCCAAGGC | GAGGAGAGTTGTAACGGCAAGG |
iNOS | GAGACAGGGAAGTCTGAAGCAC | CCAGCAGTAGTTGCTCCTCTTC |
IL1β | TGGACCTTCCAGGATGAGGACA | GTTCATCTCGGAGCCTGTAGTG |
TNF-α | GGTGCCTATGTCTCAGCCTCTT | GCCATAGAACTGATGAGAGGGAG |
CD206 | GTTCACCTGGAGTGATGGTTCTC | AGGACATGCCAGGGTCACCTTT |
CD163 | GGCTAGACGAAGTCATCTGCAC | CTTCGTTGGTCAGCCTCAGAGA |
Arg-1 | CATTGGCTTGCGAGACGTAGAC | GCTGAAGGTCTCTTCCATCACC |
ALP | CCAGAAAGACACCTTGACTGTGG | TCTTGTCCGTGTCGCTCACCAT |
Runx-2 | CCTGAACTCTGCACCAAGTCCT | TCATCTGGCTCAGATAGGAGGG |
COL1α1 | CCTCAGGGTATTGCTGGACAAC | CAGAAGGACCTTGTTTGCCAGG |
OCN | GCAATAAGGTAGTGAACAGACTCC | CCATAGATGCGTTTGTAGGCGG |
GAPDH | CATCACTGCCACCCAGAAGACTG | ATGCCAGTGAGCTTCCCGTTCAG |
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Yu, S.; Zhang, Q.; Hu, M.; Zhao, B.; Liu, Z.; Li, C.; Zhang, X. Study on Optimizing Novel Antimicrobial Peptides with Bifunctional Activity to Prevent and Treat Peri-Implant Disease. Antibiotics 2022, 11, 1482. https://doi.org/10.3390/antibiotics11111482
Yu S, Zhang Q, Hu M, Zhao B, Liu Z, Li C, Zhang X. Study on Optimizing Novel Antimicrobial Peptides with Bifunctional Activity to Prevent and Treat Peri-Implant Disease. Antibiotics. 2022; 11(11):1482. https://doi.org/10.3390/antibiotics11111482
Chicago/Turabian StyleYu, Shuipeng, Qian Zhang, Meilin Hu, Borui Zhao, Zhiyang Liu, Changyi Li, and Xi Zhang. 2022. "Study on Optimizing Novel Antimicrobial Peptides with Bifunctional Activity to Prevent and Treat Peri-Implant Disease" Antibiotics 11, no. 11: 1482. https://doi.org/10.3390/antibiotics11111482