Molecular Diagnosis and Therapy of Dental Caries by Oral Microbiome-Selective Aggregation-Induced Photosensitivity

Abstract

Dental caries represents one of the most widespread oral bacterial infections, affecting billions of individuals worldwide and presenting significant public health challenges. Effective diagnosis and treatment are hindered by the limitations of traditional clinical treatment methodologies, which often involve laborious examinations and invasive procedures. In this study, we employ an aggregation-induced emission probe, MeOTpy, for instant molecular detection and photodynamic treatment of dental caries. MeOTpy interacts specifically with cariogenic bacteria, exhibiting bright fluorescence upon binding to bacteria and generating reactive oxygen species under white light irradiation. This aggregation-induced photosensitivity enables rapid assessment of carious disease through fluorescent detection in dental plaque samples, facilitating easy identification of lesion sites in decayed teeth treated with MeOTpy. Furthermore, photodynamic inhibition of cariogenic biofilms is achieved by culturing ex vivo biofilms isolated from children with severe early childhood caries. MeOTpy also effectively prevents dental caries while preserving oral microbial diversity in a cariogenic infection model on rat pups. This study presents an integrated strategy for the precise diagnosis and treatment of dental caries at the point of care.

Keywords

aggregation-induced emission / biofilm / caries / molecular diagnosis / oral microbiome / photodynamic therapy

References

[1] N. B. Pitts, D. T. Zero, P. D. Marsh, et al., “Dental Caries,” Nature reviews Disease primers 3 (2017): 1-16.

[2] R. H. Selwitz, A. I. Ismail, and N. B. Pitts, “Dental Caries,” The Lancet 369, no. 9555 (2007): 51-59.

[3] T. Biswas, “India Launches Programme for Child-health Screening,” The Lancet 381, no. 9869 (2013): e8.

[4] D. Crosser, “Oral Health: Oft Overlooked,” Lancet Child Adolesc Health 3, no. 10 (2019): 663.

[5] H. Cho, Z. Ren, K. Divaris, et al., “Selenomonas sputigena Acts as a Pathobiont Mediating Spatial Structure and Biofilm Virulence in Early Childhood Caries,” Nature Communications 14, no. 1 (2023): 2919.

[6] J. Zou, M. Meng, C. S. Law, Y. Rao, and X. Zhou, “Common Dental Diseases in Children and Malocclusion,” International Journal of Oral Science 10 (2018): 7.

[7] L. Cheng, L. Zhang, L. Yue, et al., “Expert Consensus on Dental Caries Management,” International Journal of Oral Science 14, no. 1 (2022): 17.

[8] X. Zhou, Dental Caries: Principles and Management (2016): 1-183.

[9] T. Walsh, J. M. Oliveira-Neto, and D. Moore, “Chlorhexidine Treatment for the Prevention of Dental Caries in Children and Adolescents,” Cochrane Database of Systematic Reviews 13, no. 4 (2015): CD008457.

[10] J. M. Ten Cate, “Novel Anticaries and Remineralizing Agents: Prospects for the Future,” Journal of Dental Research 91, no. 9 (2012): 813-815.

[11] J. L. Baker, X. He, and W. Shi, “Precision Reengineering of the Oral Microbiome for Caries Management,” Advances in Dental Research 30, no. 2 (2019): 34-39.

[12] B. Cao, Y. Ma, J. Zhang, et al., “Oxygen Self-Sufficient Nanodroplet Composed of Fluorinated Polymer for High-Efficiently PDT Eradicating Oral Biofilm,” Materials Today Bio 26 (2024): 101091.

[13] X. Li, W. Wang, Q. Gao, et al., “Intelligent Bacteria-targeting ZIF-8 Composite for Fluorescence Imaging-guided Photodynamic Therapy of Drug-resistant Superbug Infections and Burn Wound Healing,” Exploration 4 (2024): 20230113.

[14] S. Guo, H. Tang, Y. Zhang, Z. Wang, and S. C. Tan, “Recent Advances in Biomimetic Aggregation-induced Emission Photosensitizers for Photodynamic Therapy and Immunotherapy,” Biomedical Engineering Materials 2 (2024): e12076.

[15] Y. Gao, Y. Li, Z. Xu, S. Yu, J. Liu, and H. Sun, “Multiporphyrinic Architectures: Advances in Structural Design for Photodynamic Therapy,” Aggregate 5, no. 1 (2024): e420.

[16] X. Hu, H. Zhang, Y. Wang, et al., “Synergistic Antibacterial Strategy Based on Photodynamic Therapy: Progress and Perspectives,” Journal of Chemical Engineering 450 (2022): 138129.

[17] Y. Zhou, C. Wu, X. Wang, et al., “Exploring the Changes of Peroxisomal Polarity in the Liver of Mice With Nonalcoholic Fatty Liver Disease,” Analytical Chemistry 93, no. 27 (2021): 9609-9620.

[18] T. Li, Y. Wu, W. Cai, et al., “Vision Defense: Efficient Antibacterial AIEgens Induced Early Immune Response for Bacterial Endophthalmitis,” Advanced Science 9, no. 25 (2022): 2202485.

[19] J. Li, Z. Meng, Z. Zhuang, et al., “Effective Therapy of Drug-resistant Bacterial Infection by Killing Planktonic Bacteria and Destructing Biofilms With Cationic Photosensitizer Based on Phosphindole Oxide,” Small 18, no. 17 (2022): 2200743.

[20] M. C. Giel and Y. Hong, “Click Chemistry in the Design of AIEgens for Biosensing and Bioimaging,” Aggregate 4, no. 4 (2023): e336.

[21] W. H. Bowen, R. A. Burne, H. Wu, and H. Koo, “Oral Biofilms: Pathogens, Matrix, and Polymicrobial Interactions in Microenvironments,” Trends in Microbiology 26, no. 3 (2018): 229-242.

[22] Y. Liu, L. Shi, L. Su, et al., “Nanotechnology-based Antimicrobials and Delivery Systems for Biofilm-infection Control,” Chemical Society Reviews 48, no. 2 (2019): 428-446.

[23] S. M. Alaki, B. A. Burt, and S. L. Garetz, “The Association Between Antibiotics Usage in Early Childhood and Early Childhood Caries,” Pediatric Dentistry 31, no. 1 (2009): 31.

[24] M. Sanz, D. Beighton, M. A. Curtis, et al., “Role of Microbial Biofilms in the Maintenance of Oral Health and in the Development of Dental Caries and Periodontal Diseases. Consensus Report of Group 1 of the Joint EFP/ORCA Workshop on the Boundaries Between Caries and Periodontal Disease,” Journal of Clinical Periodontology 44 (2017): S5-S11.

[25] A. S. Hemadi, R. Huang, Y. Zhou, and J. Zou, “Salivary Proteins and Microbiota as Biomarkers for Early Childhood Caries Risk Assessment,” International Journal of Oral Science 9, no. 11 (2017): e1.

[26] H. Wang, Q. Li, P. Alam, et al., “Aggregation-Induced Emission (AIE), Life and Health,” ACS Nano 17, no. 15 (2023): 14347-14405.

[27] J. Guan, R. Wei, A. Prlj, et al., “Direct Observation of Aggregation-Induced Emission Mechanism,” Angewandte Chemie International Edition 59, no. 35 (2020): 14903-14909.

[28] X. Cai and B. Liu, “Aggregation-induced Emission: Recent Advances in Materials and Biomedical Applications,” Angewandte Chemie International Edition 59, no. 25 (2020): 9868-9886.

[29] M. M. S. Lee, D. Yan, J. H. C. Chau, et al., “Highly Efficient Phototheranostics of Macrophage-Engulfed Gram-positive Bacteria Using a NIR Luminogen with Aggregation-induced Emission Characteristics,” Biomaterials (2020): 261.

[30] M. Kang, C. Zhou, S. Wu, et al., “Evaluation of Structure-function Relationships of Aggregation-induced Emission Luminogens for Simultaneous Dual Applications of Specific Discrimination and Efficient Photodynamic Killing of Gram-positive Bacteria,” Journal of the American Chemical Society 141, no. 42 (2019): 16781-16789.

[31] L. Wang, R. Chen, G. Q. Han, et al., “Super-Resolution Analyzing Spatial Organization of Lysosomes With an Organic Fluorescent Probe,” Exploration 2, no. 3 (2022): 20210215.

[32] J. Mei, Y. Huang, and H. Tian, “Progress and Trends in AIE-Based Bioprobes: A Brief Overview,” ACS Applied Matererial Interfaces 10, no. 15 (2018): 12217-12261.

[33] Z. Zhao, H. Zhang, J. W. Y. Lam, and B. Z. Tang, “Aggregation-induced Emission: New Vistas at the Aggregate Level,” Angewandte Chemie International Edition 59, no. 25 (2020): 9888-9907.

[34] Q. Peng and Z. Shuai, “Molecular Mechanism of Aggregation-induced Emission,” Aggregate 2, no. 5 (2021): e91.

[35] C. Wu, Y. Mao, X. Qi, et al., “Tracking Interactions Between TAMs and CAFs Mediated by Arginase-induced Proline Production During Immune Evasion of HCC,” Aggregate 5 (2024): e530.

[36] T. Zhang, X. Chen, C. Yuan, et al., “Near-infrared Aggregation-induced Emission Luminogens for in Vivo Theranostics of alzheimer's Disease,” Angewandte Chemie International Edition 62, no. 2 (2023): e202211550.

[37] F. Sun, W. Zhao, H. Shen, et al., “Design of Smart Aggregates: Toward Rapid Clinical Diagnosis of Hyperlipidemia in human Blood,” Advanced Materials 34, no. 49 (2022): 2207671.

[38] P. Cen, J. Huang, C. Jin, et al., “Aggregation-induced Emission Luminogens for in Vivo Molecular Imaging and Theranostics in Cancer,” Aggregate 4, no. 5 (2023): e352.

[39] H. Li, B. Q. Jin, Y. Wang, B. Deng, D. Wang, and B. Z. Tang, “As fiber Meets With AIE: Opening a Wonderland for Smart Flexible Materials,” Advanced Materials 35 (2022): 2210085.

[40] E. P. Parrott, N. Y. Tan, R. Hu, J. A. Zeitler, B. Z. Tang, and E. Pickwell-MacPherson, “Direct Evidence to Support the Restriction of Intramolecular Rotation Hypothesis for the Mechanism of Aggregation-induced Emission: Temperature Resolved Terahertz Spectra of Tetraphenylethene,” Materials Horizons 1, no. 2 (2014): 251-258.

[41] X. Hu, D. Wang, Y. Wang, Y. Wang, and S. Zhang, “Ultrafast Restricted Intramolecular Rotation in Molecules With Aggregation Induced Emission,” Journal of Chemical Physics 160, no. 15 (2024): 154302.

[42] H. Q. Yin, X. Y. Wang, and X. B. Yin, “Rotation Restricted Emission and Antenna Effect in Single Metal-organic Frameworks,” Journal of the American Chemical Society 141, no. 38 (2019): 15166-15173.

[43] S. Benedetti, S. Catalani, S. De Stefani, et al., “A Microplate-based DCFH-DA Assay for the Evaluation of Oxidative Stress in Whole Semen,” Heliyon 8, no. 9 (2022): e10642.

[44] C. Flors, M. J. Fryer, J. Waring, et al., “Imaging the Production of Singlet Oxygen in Vivo Using a New Fluorescent Sensor, Singlet Oxygen Sensor Green (R),” Journal of Experimental Botany 57, no. 8 (2006): 1725-1734.

[45] X. Liu, L. Shen, W. Xu, et al., “Low Frequency Hydromechanics-driven Generation of Superoxide Radicals via Optimized Piezotronic Effect for Water Disinfection,” Nano Energy 88 (2021): 106290.

[46] J. H. Chenand and C. T. Ho, “Antioxidant Activities of Caffeic Acid and Its Related Hydroxycinnamic Acid Compounds,” Journal of Agricultural and Food Chemistry 45, no. 7 (1997): 2374-2378.

[47] X. Zhou and K. Mopper, “Determination of Photochemically Produced Hydroxyl Radicals in Seawater and Freshwater,” Marine Chemistry 30 (1990): 71-88.

[48] H. Bai, H. Yuan, C. Nie, et al., “A Supramolecular Antibiotic Switch for Antibacterial Regulation,” Angewandte Chemie International Edition 54, no. 45 (2015): 13208-13213.

[49] R. J. Lamont, H. Koo, and G. Hajishengallis, “The Oral Microbiota: Dynamic Communities and Host Interactions,” Nature Reviews Microbiology 16, no. 12 (2018): 745-759.

[50] N. Takahashi and B. Nyvad, “The Role of Bacteria in the Caries Process: Ecological Perspectives,” Journal of Dental Research 90, no. 3 (2011): 294-303.

[51] A. Priya, C. B. M. Kumar, A. Valliammai, A. Selvaraj, and S. K. Pandian, “Usnic Acid Deteriorates Acidogenicity, Acidurance and Glucose Metabolism of Streptococcus Mutans Through Downregulation of Two-component Signal Transduction Systems,” Scientific reports 11, no. 1 (2021): 1374.

[52] Y. Liu, P. C. Naha, G. Hwang, et al., “Topical Ferumoxytol Nanoparticles Disrupt Biofilms and Prevent Tooth Decay in Vivo via Intrinsic Catalytic Activity,” Nature Communications 9 (2018): 2920.

[53] D. Kim, J. P. Barraza, R. A. Arthur, et al., “Spatial Mapping of Polymicrobial Communities Reveals a Precise Biogeography Associated With human Dental Caries,” Proceedings of the National Academy of Sciences of the United States of America 117, no. 22 (2020): 12375-12386.

[54] N. N. Fan, Y. Q. Zhou, L. J. Wu, et al., “Probing Iron-Mediated Synergistic Change of Cl⁻ and HClO in Liver Cancer Cells With a Dual-Color Fluorescence Reporter,” Analytical Chemistry 94, no. 30 (2022): 10659-10668.

[55] L. Qiao, Y. Shen, S. Zhang, et al., “H2O2-responsive Multifunctional Nanocomposite for the Inhibition of Amyloid-β and Tau Aggregation in Alzheimer's Disease,” Biomedical Engineering Materials 1, no. 1 (2023): e12011.

[56] W. Shao, F. Zhao, J. Xue, and L. Huang, “NIR-II Absorbing Organic Nanoagents for Photoacoustic Imaging and Photothermal Therapy,” Biomedical Engineering Materials 1, no. 1 (2023): e12009.

[57] P. C. Naha, Y. Liu, G. Hwang, et al., “Dextran-coated Iron Oxide Nanoparticles as Biomimetic Catalysts for Localized and pH-activated Biofilm Disruption,” ACS Nano 13, no. 5 (2019): 4960-4971.

[58] X. Yang, J. Shao, Y. Zhang, T. Wang, S. Ge, and J. Li, “Microenvironment-driven Fenton Nanoreactor Enabled by Metal-phenolic Encapsulation of Calcium Peroxide for Effective Control of Dental Caries,” Advanced Healthcare Materials 13, no. 10 (2024): 2303466.

[59] S. Hamada and H. D. Slade, “Biology, Immunology, and Cariogenicity of Streptococcus Mutans,” Microbiological Reviews 44, no. 2 (1980): 331-384.

[60] A. C. R. Tanner, J. M. J. Mathney, R. L. Kent, et al., “Cultivable anaerobic Microbiota of Severe Early Childhood Caries,” Journal of Clinical Microbiology 49, no. 4 (2011): 1464-1474.

RIGHTS & PERMISSIONS

2025 The Author(s). Aggregate published by SCUT, AIEI, and John Wiley & Sons Australia, Ltd.