Lakshmyya Kesavalu

Background: Artificial intelligence (AI) has substantial potential to advance periodontal and peri- implant care through novel diagnostic, prognostic, and therapeutic opportunities. Nevertheless, the evidence is scattered over several fields. Objective: To provide a narrative synthesis of the recent evidence regarding applications of AI to periodontology and peri-implant medicine specifically diagnostic imaging, disease prediction, biomarker discovery, peri-implant disease assessment and clinical decision support. Methods: A systematic search of PubMed, MEDLINE, Scopus, and Embase for studies from August 2020 to August 2025 was performed. We included only original research articles on AI applications in periodontal or peri-implant settings. After screening the identified records and conducting full-text assessments, a total of 32 studies were included based on established inclusion criteria. Results: AI showed high diagnostic performance at imaging-based applications, with accuracy from 0.85 to 0.98 reported for the detection and quantification of alveolar bone loss using two- and three-dimensional radiographs. Using this data, machine learning (ML) models were developed for identifying potential molecular biomarkers that could support disease prediction and gain insight into biological processes. AI-aided immune profiling and risk stratification models demonstrated robust predictive capabilities in peri-implant maintenance. mHealth is one of the types of AI-based digital health devices and has succeeded in enhancing patient compliance and periodontal prognosis. Conclusions: AI has considerable promise to impact non-invasive periodontal and peri-implant diagnostics, risk-assessment and enhanced treatment-planning through advanced imaging analysis, molecular profiling as well as patient-centred digital tools. These need to be further validated, standardized, and integrated into clinical care before widespread uptake.

Periodontitis (PD) is a polymicrobial dysbiotic immuno-inflammatory disease. Toll-like receptors (TLRs) are present on gingival epithelial cells and recognize pathogen-associated molecular patterns on pathogenic bacteria, inducing the secretion of proinflammatory cytokines and initiating innate and adaptive antigen-specific immune responses to eradicate the invading microbes. Since PD is a chronic inflammatory disease, TLR2/TLR4 play a vital role in disease pathogenesis and in maintaining the periodontium during health. Many factors modulate the TLR-mediated signaling pathway, including specific microRNAs (miRNAs). The present study was designed to characterize the function of TLR2/4 signaling in the miRNA profile after polybacterial infection with Streptococcus gordonii, Fusobacterium nucleatum, Porphyromonas gingivalis, Treponema denticola, and Tannerella forsythia in C57BL6/J wild-type, global knockout strains of TLR2 ^-/- , and TLR4 ^-/- mice ( n = 16/group) using RT-qPCR. The selection of 15 dominant miRNAs for RT-qPCR analysis was based on prior NanoString global miRNA expression profiling in response to polymicrobial and monobacterial infection. Polybacterial infections established gingival colonization in wild-type, TLR2 ^-/- , and TLR4 ^-/- mice with the induction of bacterial-specific IgG. A significant reduction in alveolar bone resorption and gingival inflammation was observed in the mandibles of TLR2/4 ^-/- mice compared to C57BL6/J wild-type mice ( P < 0.0001). Periodontal bacteria disseminated from gingival tissue to multiple organs in wild-type and TLR2 ^-/- mice (heart, lungs, brain, and kidneys) and were limited to the heart ( F. nucleatum ), lungs ( P. gingivalis ), and kidneys ( T. forsythia ) in TLR4 ^-/- mice. The diagnostic potential of miRNAs was assessed by receiver operating characteristic curves. Among the 15 miRNAs, three were upregulated in C57BL6/J wild-type mice, two in TLR2 ^-/- , and seven in TLR4 ^-/- mice. Notably, the anti-inflammatory miR-146a-5p was consistently upregulated in wild-type and TLR2 ^-/- mice but not in TLR4 ^-/- mice. Additionally, miR-15a-5p was upregulated in wild-type and TLR2 ^-/- mice. let-7c-5p was upregulated in TLR4 ^-/- mice and downregulated in wild-type mice. The upregulated miRNAs (miR-146a, miR-15a-5p, and let-7c-5p) and downregulated miRNAs could be candidate biomarkers for oral bacteria and TLRs-mediated induction of periodontitis if they are confirmed in large-scale human studies. Multi-species oral bacterial infection alters the TLR2/4 signaling pathways by modulating the expression of several potential biomarker miRNAs in the periodontium. IMPORTANCE: Periodontitis is the most prevalent chronic immuno-infectious multispecies dysbiotic disease of the oral cavity. The Toll-like receptors (TLRs) provide the first line of defense, one of the best-characterized pathogen detection systems, and play a vital role in recognizing multiple microbial products. Multispecies infection with periodontal bacteria Streptococcus gordonii , Fusobacterium nucleatum , Porphyromonas gingivalis , Treponema denticola , and Tannerella forsythia induced gingival inflammation, alveolar bone resorption (ABR), and microRNA (miRNA) expression in the C57BL6/J wild-type mice, whereas infection did not significantly increase ABR in the TLR2/4 deficient mice. Among the 15 miRNAs investigated, miR-146a-5p and miR-15a-5p were upregulated in wild-type and TLR2 ^-/- mice, and miR-146a-5p, miR-30c-5p, and let-7c-5p were upregulated in the TLR4 ^-/- mice compared to sham-infected controls. Notably, the inflammatory miRNA miR-146a-5p was uniquely upregulated among the wild-type and TLR2 ^-/- mice but not in the TLR4 ^-/- mice. The upregulated miRNAs (miR-146a, miR-15-a-5p, let-7c-5p) and downregulated miRNAs could be candidate biomarkers for oral bacteria and TLRs-mediated induction of periodontitis if they are confirmed in large-scale human studies.

Fusobacterium nucleatum ( F. nucleatum ) is an oral commensal bacterium that can become pathogenic and is associated with periodontitis, adverse pregnancy outcomes, and colorectal cancer (CRC). MicroRNAs are conserved, non-coding RNA molecules that regulate gene expression and are detected in microbial infections. This study aims to characterize the microRNA expression kinetics in the mandibles of C57BL/6J mice infected with F. nucleatum for 8 and 16 weeks and to identify miRNAs as potential biomarkers using NanoString nCounter miRNA panels. Mice were divided into four groups: 8 weeks of infection, 16 weeks of infection, and their respective sham infection. F. nucleatum -infected mice showed 100% bacterial colonization on the gingival surface, along with a significant increase in alveolar bone resorption ( P < 0.0001) and intravascular dissemination to the heart, indicating its invasive potential. Out of 577 miRNAs analyzed, seven miRNAs were upregulated, and two miRNAs were downregulated in the 8 weeks of infection group. In the 16 weeks of infection group, seven miRNAs were upregulated while 13 miRNAs were downregulated. Notably, miR-205, miR-210, and miR-199a-3p were differentially expressed at 8 weeks as well as miR-28 at 16 weeks and have been previously reported in human periodontitis. The 13 miRNAs induced by F. nucleatum (e.g., miR-361-5p) are linked to 13 multiple malignancies. In addition, miR-126-5p has been identified as a potential biomarker for patients with PD and cardiovascular disease. These results indicate that F. nucleatum induces several PD-related miRNAs and links them to systemic comorbidities. Furthermore, this study revealed F. nucleatum induction of 14 oncogenic miRNAs, and specifically, 12 miRNAs were linked with CRC. IMPORTANCE: Our study investigated oral commensal Fusobacterium nucleatum , a critical bacterium associated with gum disease, adverse pregnancy outcomes, and enriched several tumors, including colorectal cancer (CRC). Recently, microRNAs have emerged as critical players in the interactions between host and microbes, and many host functions have been reported to be regulated by miRNAs during infection. F. nucleatum oral infection in mice induced gum disease, disseminated to the heart, lungs, and several miRNAs. Elevated miR-361 expression was linked to multiple cancers. In addition, miR-126-5p expression has been reported as a potential biomarker in patients with periodontitis and coronary artery disease, indicating F. nucleatum ’s virulence potential. The 13 miRNAs induced by F. nucleatum are linked to 13 multiple malignancies, including CRC. These results indicate that F. nucleatum acts as a potent cancer-causing bacterium. This study opens new avenues for exploring F. nucleatum ’s role in gum disease and its link with cancer.

Periodontitis (PD) is a polymicrobial dysbiotic immuno-inflammatory disease. Toll-like receptors (TLRs) are present on gingival epithelial cells and recognize pathogen-associated molecular patterns (PAMPs) on pathogenic bacteria, induce the secretion of proinflammatory cytokines, and initiate innate and adaptive antigen-specific immune responses to eradicate the invading microbes. Since PD is a chronic inflammatory disease, TLR2/TLR4 plays a vital role in disease pathogenesis and maintaining the periodontium during health. Many factors modulate the TLR-mediated signaling pathway, including specific miRNAs. The present study was designed to characterize the function of TLR2/4 signaling to the miRNA profile after polybacterial infection with Streptococcus gordonii, Fusobacterium nucleatum, Porphyromonas gingivalis, Treponema denticola, and Tannerella forsythia in C57BL6/J wild-type, TLR2 ^−/− , and TLR4 ^−/− mice (n=16/group) using RT-qPCR. The selection of 15 dominant miRNAs for RT-qPCR analysis was based on prior NanoString global miRNA expression profiling in response to polymicrobial and monobacterial infection. Polybacterial infections established gingival colonization in wild-type, TLR2 ^−/− and TLR4 ^−/− mice with induction of bacterial-specific IgG. A significant reduction in alveolar bone resorption (ABR) and gingival inflammation was observed in the mandibles of TLR2/4 ^−/− mice compared to C57BL6/J wild-type mice ( p −/− mice (heart, lungs, brain, kidney) and limited to heart ( F. nucleatum ), lungs ( P. gingivalis ), kidney ( T. forsythia ) in TLR4 ^−/− mice. The diagnostic potential of miRNAs was assessed by receiver operating characteristic (ROC) curves. Among 15 miRNAs, three were upregulated in C57BL6/J wild-type mice, two in TLR2 ^−/− , and seven in TLR4 ^−/− mice. Notably, the anti-inflammatory miR-146a-5p was consistently upregulated in all the mice. Additionally, miR-15a-5p was upregulated in wild-type and TLR2 ^−/− mice. let-7c-5p was upregulated in TLR4 ^−/− mice and downregulated in the wild-type mice. Multi-species oral bacterial infection alters the TLR2/4 signaling pathways by modulating the expression of several potential biomarker miRNAs in periodontium. IMPORTANCE: Periodontitis is the most prevalent chronic immuno-infectious multispecies dysbiotic disease of the oral cavity. The Toll-like receptors (TLR) provide the first line of defense, one of the best-characterized pathogens-detection systems and play a vital role in recognizing multiple microbial products. Multispecies infection with periodontal bacteria S. gordonii, F. nucleatum, P. gingivalis, T. denticola, and T. forsythia induced gingival inflammation, alveolar bone resorption (ABR) and miRNA expression in the C57BL6/J wild-type mice and whereas infection did not increase significant ABR in the TLR2/4 deficient mice. Among the 15 miRNAs investigated, miR-146a - 5p, miR-15a-5p were upregulated in wild-type and TLR2 ^−/− mice and miR-146a-5p, miR-30c-5p, let-7c-5p were upregulated in the TLR4 ^−/− mice compared to sham-infected controls. Notably, inflammatory miRNA miR-146a-5p was upregulated uniquely among the three different infection groups. The upregulated miRNAs (miR-146a, miR-15-a-5p, let-7c-5p) and downregulated miRNAs could be markers for TLRs-mediated induction of periodontitis.

Fusobacterium nucleatum (F. nucleatum, Fn), an oral commensal that can become pathogenic, is associated with periodontitis (PD), adverse pregnancy outcomes, and colorectal cancer (CRC). MicroRNAs (miRNAs) are conserved, non-coding RNAs that play a regulatory role in gene expression and are detected in microbial infections. The aim of this study was to characterize the global microRNA expression kinetics in the mandibles of C57BL/6J mice infected with F. nucleatum for 8 and 16 weeks, and to identify miRNAs as biomarkers of the PD process using high-throughput NanoString nCounter® miRNA panels. Mice were divided into four groups: 8 weeks of Fn infection, 16 weeks of Fn infection, and their respective sham infection. Fn-infected mice at both 8- and 16 weeks showed 100% bacterial colonization on the gingival surface, along with a significant increase in alveolar bone resorption (p<0.0001) and intravascular dissemination to heart, indicating its invasive potential. Out of 577 miRNAs analyzed, seven miRNAs (miR-361-5p, miR-99b-5p) were upregulated, and two miRNAs (miR-362-3p and miR-720) were downregulated in the 8-week Fn infection group. In the 16-week Fn infection group, seven miRNAs (miR-361-5p, miR-99b-5p) were upregulated miRNAs and 13 miRNAs (miR-323-3p, miR-488) were downregulated. Notably, miRNAs such as miR-205, miR-210, miR-199a-3p which were differentially expressed (DE) in the Fn-infected mice at 8 weeks and miR-28 at 16 weeks, have been previously reported in human periodontitis. The 13 miRNAs induced by F. nucleatum (miR-361-5p) are linked to multiple malignancies, including esophageal, gastric, pancreatic, and colorectal cancers. Additionally, the DE miR-126-5p in the 8-week infection group has been identified as a potential biomarker for patients with PD and cardiovascular disease. The results indicate that F. nucleatum acts as a potent oncomicrobe, inducing several miRNAs associated with PD and linking it to systemic comorbidities. Furthermore, the study revealed similar expressions between PD-associated miRNAs and nine CRC tumor-expressed miRNAs.

Persistent inflammation is a major contributor in the development of various inflammatory diseases like atherosclerosis. Our study investigates how transient receptor potential vanilloid 4 (TRPV4), a mechanosensitive ion channel, interacts with microRNA-146a (miR-146a), within the context of inflammation and atherosclerosis. Micro-RNAs play a critical role in controlling gene expression, and miR-146a is notable for its anti-inflammatory actions. TRPV4 is activated by diverse soluble and mechanical stimuli, and often associated with inflammatory responses in various diseases. Here, we find that TRPV4 negatively regulates miR-146a expression in macrophages, especially following stimulation by lipopolysaccharides or alterations in matrix stiffness. We show that in atherosclerosis, a condition characterized by matrix stiffening, TRPV4 decreases miR-146a expression in aortic tissue macrophages. We find that TRPV4’s impact on miR-146a is independent of activation of NFκB, Stat1, P38, and AKT, but is rather mediated through a mechanism involving histone deacetylation instead of DNA methylation at the miR-146a promoter site. Furthermore, we show that N-terminal residues 1 to 130 in TRPV4 is essential in suppression of miR-146a expression in LPS-stimulated macrophages. Altogether, this study identifies a regulatory mechanism of miR-146a expression by TRPV4 which may open new potential therapeutic strategies for managing inflammatory diseases.

This study aimed to comprehensively assess the influence of the nanotube diameter and the presence of a silicon carbide (SiC) coating on microbial proliferation on nanostructured titanium surfaces. An experiment used 72 anodized titanium sheets with varying nanotube diameters of 50 and 100 nm. These sheets were divided into four groups: non-coated 50 nm titanium nanotubes, SiC-coated 50 nm titanium nanotubes, non-coated 100 nm titanium nanotubes, and SiC-coated 100 nm titanium nanotubes, totaling 36 samples per group. P. gingivalis and T. denticola reference strains were used to evaluate microbial proliferation. Samples were assessed over 3 and 7 days using fluorescence microscopy with a live/dead viability kit and scanning electron microscopy (SEM). At the 3-day time point, fluorescence and SEM images revealed a lower density of microorganisms in the 50 nm samples than in the 100 nm samples. However, there was a consistently low density of T. denticola across all the groups. Fluorescence images indicated that most bacteria were viable at this time. By the 7th day, there was a decrease in the microorganism density, except for T. denticola in the non-coated samples. Additionally, more dead bacteria were detected at this later time point. These findings suggest that the titanium nanotube diameter and the presence of the SiC coating influenced bacterial proliferation. The results hinted at a potential antibacterial effect on the 50 nm diameter and the coated surfaces. These insights contribute valuable knowledge to dental implantology, paving the way for developing innovative strategies to enhance the antimicrobial properties of dental implant materials and mitigate peri-implant infections.

Streptococcus gordonii (S. gordonii, Sg) is one of the early colonizing, supragingival commensal bacterium normally associated with oral health in human dental plaque. MicroRNAs (miRNAs) play an important role in the inflammation-mediated pathways and are involved in periodontal disease (PD) pathogenesis. PD is a polymicrobial dysbiotic immune-inflammatory disease initiated by microbes in the gingival sulcus/pockets. The objective of this study is to determine the global miRNA expression kinetics in S. gordonii DL1-infected C57BL/6J mice. All mice were randomly divided into four groups (n = 10 mice/group; 5 males and 5 females). Bacterial infection was performed in mice at 8 weeks and 16 weeks, mice were euthanized, and tissues harvested for analysis. We analyzed differentially expressed (DE) miRNAs in the mandibles of S. gordonii-infected mice. Gingival colonization/infection by S. gordonii and alveolar bone resorption (ABR) was confirmed. All the S. gordonii-infected mice at two specific time points showed bacterial colonization (100%) in the gingival surface, and a significant increase in mandible and maxilla ABR (p < 0.0001). miRNA profiling revealed 191 upregulated miRNAs (miR-375, miR-34b-5p) and 22 downregulated miRNAs (miR-133, miR-1224) in the mandibles of S. gordonii-infected mice at the 8-week mark. Conversely, at 16 weeks post-infection, 10 miRNAs (miR-1902, miR-203) were upregulated and 32 miRNAs (miR-1937c, miR-720) were downregulated. Two miRNAs, miR-210 and miR-423-5p, were commonly upregulated, and miR-2135 and miR-145 were commonly downregulated in both 8- and 16-week-infected mice mandibles. Furthermore, we employed five machine learning (ML) algorithms to assess how the number of miRNA copies correlates with S. gordonii infections in mice. In the ML analyses, miR-22 and miR-30c (8-week), miR-720 and miR-339-5p (16-week), and miR-720, miR-22, and miR-339-5p (combined 8- and 16-week) emerged as the most influential miRNAs.