S. mutans Antisense vicK RNA Over-Expression Plus Antibacterial Dimethylaminohexadecyl Methacrylate Suppresses Oral Biofilms and Protects Enamel Hardness in Extracted Human Teeth
Abstract
:1. Introduction
2. Materials and Methods
2.1. Synthesis of DMAHDM
2.2. Strain Verification and Biofilm Formation
Primers | Sequence 5′-3′ (Forward/Reverse) | Source or Reference |
---|---|---|
gyrA | 5′-ATTGTTGCTCGGGCTCTTCCAG-3′/ 5′-ATGCGGCTTGTCAGGAGTAACC-3′ | [35] |
ASvicK | 5′-GTTGATTGCTGACTTTGAATTTGA-3′/ 5′-GCGTATGATTACTGATTTACTTAGC-3′ | [13] |
vicK | 5′-CACTTTACGCATTCGTTTTGCC-3′/ 5′-CGTTCTTCTTTTTCCTGTTCGGTC-3′ | [35] |
gtfB | 5′-ACACTTTCGGGTGGCTTG-3′/ 5′-GCTTAGATGTCACTTCGGTTG-3′ | [35] |
gtfC | 5′-CCAAAATGGTATTATGGCTGTCG-3′/ 5′-TGAGTCTCTATCAAAGTAACGCAG-3′ | [35] |
gtfD | 5′-AATGAAATTCGCAGCGGACTTGAG-3′/ 5′-TTAGCCTGACGCATGTCTTCATTGTA-3′ | [35] |
ftf | 5′-ATTGGCGAACGGCGACTTACTC-3′/ 5′-CCTGCGACTTCATTACGATTGGTC-3′ | [35] |
2.3. Determining MIC, MBC-P, and MBC-B of DMAHDM
2.4. Growth Curves
2.5. Live/Dead Staining Assay
2.6. Biofilm Viability Using the MTT Assay
2.7. Colony-Forming Unit (CFU) Counts
2.8. Crystal Violet (CV) Staining
2.9. Lactic Acid Secretion
2.10. Biofilm Imaging
2.11. Polysaccharide Synthesis
2.12. Quantitative Real-Time-Polymerase-Chain Reaction (qRT-PCR)
2.13. Human Tooth Enamel Hardness Measurement
2.14. Statistical Analysis
3. Results
3.1. Synthesis of DMAHDM and Strain Verification
3.2. Susceptibility Testing of DMAHDM
3.3. Alteration of the Biofilm Formation and Lactic Acid Secretion
3.4. Inhibition of EPS Production and Biofilms Architecture
3.5. Regulation of Gene Expression Related to EPS Metabolism
3.6. Protection of Human Tooth Enamel Hardness
4. Discussion
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Strains or Plasmids | Description | Source or Reference |
---|---|---|
Parent S. mutans | Streptococcus mutans UA159 | aATCC 7000610 |
ASvicK S. mutans | ASvicK over expression strain | [13] |
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Yu, S.; Xu, M.; Wang, Z.; Deng, Y.; Xu, H.H.K.; Weir, M.D.; Homayounfar, N.; Fay, G.G.; Chen, H.; Yang, D. S. mutans Antisense vicK RNA Over-Expression Plus Antibacterial Dimethylaminohexadecyl Methacrylate Suppresses Oral Biofilms and Protects Enamel Hardness in Extracted Human Teeth. Pathogens 2024, 13, 707. https://doi.org/10.3390/pathogens13080707
Yu S, Xu M, Wang Z, Deng Y, Xu HHK, Weir MD, Homayounfar N, Fay GG, Chen H, Yang D. S. mutans Antisense vicK RNA Over-Expression Plus Antibacterial Dimethylaminohexadecyl Methacrylate Suppresses Oral Biofilms and Protects Enamel Hardness in Extracted Human Teeth. Pathogens. 2024; 13(8):707. https://doi.org/10.3390/pathogens13080707
Chicago/Turabian StyleYu, Shuang, Mengmeng Xu, Zheng Wang, Yang Deng, Hockin H. K. Xu, Michael D. Weir, Negar Homayounfar, Guadalupe Garcia Fay, Hong Chen, and Deqin Yang. 2024. "S. mutans Antisense vicK RNA Over-Expression Plus Antibacterial Dimethylaminohexadecyl Methacrylate Suppresses Oral Biofilms and Protects Enamel Hardness in Extracted Human Teeth" Pathogens 13, no. 8: 707. https://doi.org/10.3390/pathogens13080707