Journal of Dental Research

Our world is at a turning point with biological and social pathogens wreaking havoc at the same time that science and technology are exploding with new discoveries. It is a pivotal time for the new report Oral Health in America: Advances and Challenges to be released and a pivotal time for our profession to take action and lead. The art, science, and practice of dentistry is very different from 20 y ago when the original Surgeon General’s report was released. We are on the precipice of individualized health care where providers will collaborate to deliver diagnostics and therapeutics that are data driven and inclusive of the social determinants of health. To move forward with alacrity requires a strong scientific foundation, effective educational approaches, an understanding of the upstream determinants of health, and partnerships across the health professions and beyond. Oral health has never been more important, and now is the time for our profession to further develop, elevate, and translate the science into practice and policy to improve the nation’s health.

Oral potentially malignant disorders (OPMDs) are a heterogeneous group of oral lesions with a variable risk of malignant transformation to oral squamous cell carcinoma. The current OPMDs malignant transformation screening depends on conventional oral examination (COE) and is confirmed by biopsy and histologic examination. However, early malignant lesions with subtle mucosal changes are easily unnoticed by COE based on visual inspection and palpation. Optical techniques have been used to determine the biological structure, composition, and function of cells and tissues noninvasively by analyzing the changes in their optical properties. The oral epithelium and stroma undergo persistent structural, functional, and biochemical alterations during malignant transformation, leading to variations in optical tissue properties; optical techniques are thus powerful tools for detecting OPMDs malignant transformation. The optical imaging methods already used to detect OPMDs malignant transformation in vivo include autofluorescence imaging, narrowband imaging, confocal reflectance microscopy, and optical coherence tomography. They exhibit advantages over COE in detecting biochemical or morphologic changes at the molecular or cellular level in vivo; however, limitations also exist. This article comprehensively reviews the various real-time in vivo optical imaging methods used in the adjunctive diagnosis of OPMDs malignant transformation. We focus on the principles of these techniques, review their clinical application, and compare and summarize their advantages and disadvantages. Finally, we conclude with a discussion of current challenges and future directions of this field.

Taste receptors are receptor proteins that detect ligands belonging to the 5 taste modalities: sweet, bitter, sour, salty, and umami. Taste receptors are not restricted to taste cells in taste buds; rather, they are distributed throughout the entire body. For example, solitary chemosensory cells (SCCs) and tuft cells express taste signal proteins and are present in several mucosae. In the airways, SCCs sense bacteria, allergens, viruses, and noxious stimuli and drive evasive behavior, neuroinflammation, and antibacterial responses. In the gut, tuft cells detect helminth infection and bacterial dysbiosis and initiate type II immune responses characterized by tissue remodeling. In the gingiva, SCCs detect oral pathogenic bacteria, evoke innate immune responses and release antimicrobial compounds in the epithelium, and regulate the microbiome composition. This review summarizes the most recent research on extragustatory taste receptors and their function in antibacterial defense. We also discuss how these findings have provided insights into the development of potential therapeutic strategies for mucosal bacterial infection and dental diseases.

Durable resin-ceramic adhesion may influence the clinical success of ceramic restorations, which has been one of the challenging issues in dentistry. The present study assessed the bond strength and chemical interaction of 10-methacryloxydecyl dihydrogen phosphate (MDP), MDP+silane, and MDP-salt primers to alumina-blasted zirconia ceramic by tensile bond strength test, surface elemental composition with x-ray photoelectron spectroscopy analysis, contact angle measurement, surface morphology with scanning electron microscopy, and surface topography with 3-dimensional confocal laser scanning microscope analyses. MDP-salt showed the highest tensile bond strength before and after thermocycling when compared with MDP and MDP+silane (P < 0.05). The measured contact angle values differed significantly (P < 0.001) in the order of MDP-salt > control (no chemical pretreatment) > MDP+silane > MDP. There was no difference in surface roughness (P = 0.317) and surface topography among all tested groups. Zirconia treated with MDP-salt showed phosphorus peaks in addition to zirconia and alumina peaks. MDP-salt has zirconia priming properties, which improves bonding performance to resin cement.

The dentin collagen matrix that is not completely enveloped by resin adhesive is vulnerable to degradation by intrinsic collagenases during the etch-and-rinse process, which contributes to the deterioration of the bonding interface. Current commercial adhesives have no functional components that can form covalent bonds to the dentin collagen matrix. In this study, a photocurable aldehyde, 4-formylphenyl acrylate (FA), was synthesized and for the first time applied as a primer in adhesive dentistry to covalently bind to collagen. Experimental groups with different concentrations of FA (1%, 3%, 5%, 7%, 9%) were prepared as primers. The cytotoxicity was evaluated by live/dead-cell staining and thiazolyl blue tetrazolium bromide assay. The interaction of FA with collagen was examined by attenuated total reflection Fourier transform infrared spectroscopy, hydroxyproline release under the degradation of type I collagenase, and thermogravimetric analysis. An optimal group was selected based on the degree of conversion of 2 universal adhesives and further divided depending on the treatment time (20 s, 30 s, 1 min, 2 min). The bonding performances were evaluated by microtensile strength before and after aging. Finally, the bonding interface was observed under confocal laser scanning microscopy and scanning electron microscope. The results indicated that FA demonstrated good biocompatibility, dentin modification capability, and infiltration. It not only effectively cross-linked dentin collagen to improve its stability against enzymatic hydrolysis and modify the adhesive interface but also potentially acted as a diluting monomer to induce deep penetration of adhesive resin monomers into the dentin. The bonding strength after aging was improved without jeopardizing the degree of conversion of 2 commercial adhesives. Such prominent advantages of using FA to improve the bonding performance promotes its further application in adhesive dentistry.

Many dental procedures are considered aerosol-generating procedures that may put the dental operator and patients at risk for cross-infection due to contamination from nasal secretions and saliva. This aerosol, depending on the size of the particles, may stay suspended in the air for hours. The primary objective of the study was to characterize the size and concentrations of particles emitted from 7 different dental procedures, as well as estimate the contribution of the nasal and salivary fluids of the patient to the microbiota in the emitted bioaerosol. This cross-sectional study was conducted in an open-concept dental clinic with multiple operators at the same time. Particle size characterization and mass and particle concentrations were done by using 2 direct reading instruments: Dust-Trak DRX (Model 8534) and optical particle sizer (Model 3330). Active bioaerosol sampling was done before and during procedures. Bayesian modeling (SourceTracker2) of long-reads of the 16S ribosomal DNA was used to estimate the contribution of the patients’ nasal and salivary fluids to the bioaerosol. Aerosols in most dental procedures were sub-PM₁ dominant. Orthodontic debonding and denture adjustment consistently demonstrated more particles in the PM₁, PM_2.5, PM₄, and PM₁₀ ranges. The microbiota in bioaerosol samples were significantly different from saliva and nasal samples in both membership and abundance (P < 0.05) but not different from preoperative ambient air samples. A median of 80.15% of operator exposure was attributable to sources other than the patients’ salivary or nasal fluids. Median operator’s exposure from patients’ fluids ranged from 1.45% to 2.75%. Corridor microbiota showed more patients’ nasal bioaerosols than oral bioaerosols. High-volume saliva ejector and saliva ejector were effective in reducing bioaerosol escape. Patient nasal and salivary fluids are minor contributors to the operator’s bioaerosol exposure, which has important implications for COVID-19. Control of bioaerosolization of nasal fluids warrants further investigation.

The roles of Wnt/β-catenin signaling in regulating the morphology and microstructure of craniomaxillofacial (CMF) bones was explored using mice carrying a constitutively active form of β-catenin in activating Dmp1-expressing cells (e.g., daβcat^Ot mice). By postnatal day 24, daβcat^Ot mice exhibited midfacial truncations coupled with maxillary and mandibular hyperostosis that progressively worsened with age. Mechanistic insights into the basis for the hyperostotic facial phenotype were gained through molecular and cellular analyses, which revealed that constitutively activated β-catenin in Dmp1-expressing cells resulted in an increase in osteoblast number and an increased rate of mineral apposition. An increase in osteoblasts was accompanied by an increase in osteocytes, but they failed to mature. The resulting CMF bone matrix also had an abundance of osteoid, and in locations where compact lamellar bone typically forms, it was replaced by porous, woven bone. The hyperostotic facial phenotype was progressive. These findings identify for the first time a ligand-independent positive feedback loop whereby unrestrained Wnt/β-catenin signaling results in a CMF phenotype of progressive hyperostosis combined with architecturally abnormal, poorly mineralized matrix that is reminiscent of craniotubular disorders in humans.

Periodontal ligament derived stem cells (PDLSCs) are capable of differentiating into multiple cell types and inducing a promising immunomodulation for tissue regeneration and disease treatment. However, it is still challenging to develop a practical approach to activate endogenous stem cells for tissue self-healing and regeneration. In this study, transcriptome analysis reveals that resveratrol promotes PDLSC stemness through activation of stem cell, osteoprogenitor, and chondroprogenitor markers. Self-renewal and multipotent differentiation abilities are also improved in resveratrol-treated PDLSCs. In addition, immunomodulation of PDLSCs is dramatically increased after resveratrol treatment. Mechanistically, we show that resveratrol activates ERK/WNT crosstalk through elevation of olfactory and growth factor signaling pathways to upregulate the expression levels of RUNX2 and FASL for osteogenesis and immunomodulation, respectively. By using a periodontitis animal model, administration of resveratrol partially rescues bone loss through activation of endogenous somatic stem cells and inhibition of inflammatory T-cell infiltration. Taken together, our findings identify a novel pharmacological approach to achieve autotherapies for endogenous tissue regeneration.

Periodontitis is a highly prevalent chronic inflammatory disease that progressively destroys the structures supporting teeth, leading to tooth loss. Periodontal tissue is innervated by abundant pain-sensing primary afferents expressing neuropeptides and transient receptor potential vanilloid 1 (TRPV1). However, the roles of nociceptive nerves in periodontitis and bone destruction are controversial. The placement of ligature around the maxillary second molar or the oral inoculation of pathogenic bacteria induced alveolar bone destruction in mice. Chemical ablation of nociceptive neurons in the trigeminal ganglia achieved by intraganglionic injection of resiniferatoxin decreased bone loss in mouse models of experimental periodontitis. Consistently, ablation of nociceptive neurons decreased the number of osteoclasts in alveolar bone under periodontitis. The roles of nociceptors were also determined by the functional inhibition of TRPV1-expressing trigeminal afferents using an inhibitory designer receptor exclusively activated by designer drugs (DREADD) receptor. Noninvasive chemogenetic functional silencing of TRPV1-expressing trigeminal afferents not only decreased induction but also reduced the progression of bone loss in periodontitis. The infiltration of leukocytes and neutrophils to the periodontium increased at the site of ligature, which was accompanied by increased amount of proinflammatory cytokines, such as receptor activator of nuclear factor κΒ ligand, tumor necrosis factor, and interleukin 1β. The extents of increase in immune cell infiltration and cytokines were significantly lower in mice with nociceptor ablation. In contrast, the ablation of nociceptors did not alter the periodontal microbiome under the conditions of control and periodontitis. Altogether, these results indicate that TRPV1-expressing afferents increase bone destruction in periodontitis by promoting hyperactive host responses in the periodontium. We suggest that specific targeting of neuroimmune and neuroskeletal regulation can offer promising therapeutic targets for periodontitis supplementing conventional treatments.

The oral microbiota has been implicated in various neurologic conditions, including autism spectrum disorder (ASD), a category of neurodevelopmental disorders defined by core behavioral impairments. Recent data propose the etiopathogenetic role of intestinal microbiota in ASD. The aim of the present study was to elucidate whether the oral microbiota contributes to the pathogenesis of ASD. On the basis of microbial changes detected in the oral cavity of children with ASD, we transferred oral microbiota from donors with ASD and typical development (TD) into an antibiotic-mediated microbiota-depleted mouse model and found that the ASD microbiota is sufficient to induce ASD-like behaviors, such as impaired social behavior. Mice receiving oral microbiota from the ASD donor showed significantly different microbiota structures in their oral cavity and intestinal tract as compared with those receiving TD microbiota and those not receiving any bacterium. The prefrontal cortex of ASD microbiota recipient mice displayed an alternative transcriptional profile with significant upregulation of serotonin-related gene expression, neuroactive ligand-receptor interaction, and TGF-β signaling pathway relative to that in TD microbiota recipient mice. The expression of serotonin-related genes was significantly increased in ASD microbiota recipient mice and was associated with selective autistic behaviors and changes in abundance of specific oral microbiota, including species of Bacteroidetes [G-7], Porphyromonas, and Tannerella. Machine learning based on the causal inference method confirmed a contributing role of Porphyromonas sp. HMT 930 in ASD. Taken together, the oral microbiota of children with ASD can lead to ASD-like behaviors, differences in microbial community structures, and altered neurosignaling activities in recipient mice; this highlights the mouth-microbial-brain connections in the development of neuropathology and provides a novel strategy to fully understand the etiologic mechanism of ASD.

Tooth agenesis is a common structural birth defect in humans that results from failure of morphogenesis during early tooth development. The homeobox transcription factor Msx1 and the canonical Wnt signaling pathway are essential for “bud to cap” morphogenesis and are causal factors for tooth agenesis. Our recent study suggested that Msx1 regulates Wnt signaling during early tooth development by suppressing the expression of Dkk2 and Sfrp2 in the tooth bud mesenchyme, and it demonstrated partial rescue of Msx1-deficient molar teeth by a combination of DKK inhibition and genetic inactivation of SFRPs. In this study, we found that Sostdc1/Wise, another secreted Wnt antagonist, is involved in regulating the odontogenic pathway downstream of Msx1. Whereas Sostdc1 expression in the developing tooth germ was not increased in Msx1^−/− embryos, genetic inactivation of Sostdc1 rescued maxillary molar, but not mandibular molar, morphogenesis in Msx1^−/− mice with full penetrance. Since the Msx1^−/−;Sostdc1^−/− embryos exhibited ectopic Dkk2 expression in the developing dental mesenchyme, similar to Msx1^−/− embryos, we generated and analyzed tooth development in Msx1^−/−;Dkk2^−/− double and Msx1^−/−;Dkk2^−/−;Sostdc1^−/− triple mutant mice. The Msx1^−/−;Dkk2^−/− double mutants showed rescued maxillary molar morphogenesis at high penetrance, with a small percentage also exhibiting mandibular molars that transitioned to the cap stage. Furthermore, tooth development was rescued in the maxillary and mandibular molars, with full penetrance, in the Msx1^−/−;Dkk2^−/−;Sostdc1^−/− mice. Together, these data reveal 1) that a key role of Msx1 in driving tooth development through the bud-to-cap transition is to control the expression of Dkk2 and 2) that modulation of Wnt signaling activity by Dkk2 and Sostdc1 plays a crucial role in the Msx1-dependent odontogenic pathway during early tooth morphogenesis.

Methylglyoxal (MGO) is an important molecule derived from glucose metabolism with the capacity of attaching to collagen and generating advanced glycation end products (AGEs), which accumulate in tissues over time and are associated with aging and diseases. However, the accumulation of MGO-derived AGEs in dentin and their effect on the nanomechanical properties of dentinal collagen remain unknown. Thus, the aim of the present study was to quantify MGO-based AGEs in the organic matrix of human dentin as a function of age and associate these changes with alterations in the nanomechanical and ultrastructural properties of dentinal collagen. For this, 12 healthy teeth from <26-y-old and >50-y-old patients were collected and prepared to obtain crown and root dentin discs. Following demineralization, MGO-derived AGEs were quantified with a competitive ELISA. In addition, atomic force microscopy nanoindentation was utilized to measure changes in elastic modulus in peritubular and intertubular collagen fibrils. Finally, principal component analysis was carried out to determine aging profiles for crown and root dentin. Results showed an increased presence of MGO AGEs in the organic matrix of dentin in the >50-y-old specimens as compared with the <26-y-old specimens in crown and root. Furthermore, an increase in peritubular and intertubular collagen elasticity was observed in the >50-y-old group associated with ultrastructural changes in the organic matrix as determined by atomic force microscopy analysis. Furthermore, principal component analysis loading plots suggested different “aging profiles” in crown and root dentin, which could have important therapeutic implications in restorative and adhesive dentistry approaches. Overall, these results demonstrate that the organic matrix of human dentin undergoes aging-related changes due to MGO-derived AGEs with important changes in the nanomechanical behavior of collagen that may affect diagnostic and restorative procedures in older people.

Gasdermin E (GSDME), as the major executive protein of pyroptosis, has been considered to be linked to antitumor immunity in recent years. However, the role of GSDME in oral squamous cell carcinoma (OSCC) remains to be elucidated. Here, by using a human OSCC tissue microarray, human OSCC tissue, and Tgfbr1/Pten conditional knockout mice, we found that GSDME was strongly expressed in OSCC and that GSDME expression in primary tumors was higher than that in metastatic lymph nodes. In addition, GSDME expression in OSCC was positively related to better prognosis. Moreover, GSDME-mediated pyroptosis occurred upon stimulation with chemotherapy drugs, and functional knockdown of GSDME attenuated the cisplatin-induced antitumor effect. Consistent with these results, bioinformatic analysis indicated that GSDME expression was positively correlated with the sensitivity of a number of antitumor drugs approved by the US Food and Drug Administration. Inhibition of GSDME expression by small interfering RNA in SCC7 cells significantly increased the expression of the cancer stem cell markers, CD44 and ALDH1. Furthermore, multiplexed immunohistochemistry and flow cytometry indicated that the expression of GSDME positively correlated with tumor-infiltrating CD8⁺ T cells, granzyme B, and M1 phenotype macrophages. Collectively, these findings demonstrated that GSDME is a potential positive prognostic factor of OSCC, and GSDME-mediated pyroptosis induced by chemotherapy plays a role in antitumor response.

Craniofacial and jaw bones have unique physiological specificities when compared to axial and appendicular bones. However, the molecular profile of the jaw osteoblast (OB) remains incomplete. The present study aimed to decipher the bone site-specific profiles of transcription factors (TFs) expressed in OBs in vivo. Using RNA sequencing analysis, we mapped the transcriptome of confirmed OBs from 2 different skeletal sites: mandible (Md) and tibia (Tb). The OB transcriptome contains 709 TF genes: 608 are similarly expressed in Md-OB and Tb-OB, referred to as “OB-core”; 54 TF genes are upregulated in Md-OB, referred to as “Md-set”; and 18 TF genes are upregulated in Tb-OB, referred to as “Tb-set.” Notably, the expression of 29 additional TF genes depends on their RNA transcript variants. TF genes with no previously known role in OBs and bone were identified. Bioinformatics analysis combined with review of genetic disease databases and a comprehensive literature search showed a significant contribution of anatomical origin to the OB signatures. Md-set and Tb-set are enriched with site-specific TF genes associated with development and morphogenesis (neural crest vs. mesoderm), and this developmental imprint persists during growth and homeostasis. Jaw and tibia site-specific OB signatures are associated with craniofacial and appendicular skeletal disorders as well as neurocristopathies, dental disorders, and digit malformations. The present study demonstrates the feasibility of a new method to isolate pure OB populations and map their gene expression signature in the context of OB physiological environment, avoiding in vitro culture and its associated biases. Our results provide insights into the site-specific developmental pathways governing OBs and identify new major OB regulators of bone physiology. We also established the importance of the OB transcriptome as a prognostic tool for human rare bone diseases to explore the hidden pathophysiology of craniofacial malformations, among the most prevalent congenital defects in humans.