Oral Mucosal Diseases Research Articles

Machine Learning-Based Genome-Wide Salivary DNA Methylation Analysis for Identification of Noninvasive Biomarkers in Oral Cancer Diagnosis.

Simple SummaryBecause tissue biopsy is the gold standard for diagnosing oral cancer, it is often performed to confirm disease during screening, management, and monitoring. However, many reports are negative. Salivary biomarkers can provide the preliminary stratification of suspicious lesions to encourage patient selection in clinical practice. However, the discovery and implementation of salivary biomarkers still need to be refined. Therefore, in this study, we successfully utilized machine learning techniques to select optimal methylome biomarkers that may be applied for oral cancer diagnoses.This study aims to examine the feasibility of ML-assisted salivary-liquid-biopsy platforms using genome-wide methylation analysis at the base-pair and regional resolution for delineating oral squamous cell carcinoma (OSCC) and oral potentially malignant disorders (OPMDs). A nested cohort of patients with OSCC and OPMDs was randomly selected from among patients with oral mucosal diseases. Saliva samples were collected, and DNA extracted from cell pellets was processed for reduced-representation bisulfite sequencing. Reads with a minimum of 10× coverage were used to identify differentially methylated CpG sites (DMCs) and 100 bp regions (DMRs). The performance of eight ML models and three feature-selection methods (ANOVA, MRMR, and LASSO) were then compared to determine the optimal biomarker models based on DMCs and DMRs. A total of 1745 DMCs and 105 DMRs were identified for detecting OSCC. The proportion of hypomethylated and hypermethylated DMCs was similar (51% vs. 49%), while most DMRs were hypermethylated (62.9%). Furthermore, more DMRs than DMCs were annotated to promoter regions (36% vs. 16%) and more DMCs than DMRs were annotated to intergenic regions (50% vs. 36%). Of all the ML models compared, the linear SVM model based on 11 optimal DMRs selected by LASSO had a perfect AUC, recall, specificity, and calibration (1.00) for OSCC detection. Overall, genome-wide DNA methylation techniques can be applied directly to saliva samples for biomarker discovery and ML-based platforms may be useful in stratifying OSCC during disease screening and monitoring.

Electrospun patch delivery of anti-TNFα F(ab) for the treatment of inflammatory oral mucosal disease.

Chronic ulcerative oral mucosal inflammatory diseases, including oral lichen planus and recurrent aphthous stomatitis, are painful and highly prevalent, yet lack effective clinical management. In recent years, systemic biologic therapies, including monoclonal antibodies that block the activity of cytokines, have been increasingly used to treat a range of immune-mediated inflammatory conditions such as rheumatoid arthritis and psoriasis. The ability to deliver similar therapeutic agents locally to the oral epithelium could radically alter treatment options for oral mucosal inflammatory diseases, where pro-inflammatory cytokines, in particular tumour-necrosis factor-α (TNFα), are major drivers of pathogenesis. To address this, an electrospun dual-layer mucoadhesive patch comprising medical-grade polymers was investigated for the delivery of F(ab) biologics to the oral mucosa. A fluorescent-labelled F(ab) was incorporated into mucoadhesive membranes using electrospinning with 97% v/v ethanol as a solvent. The F(ab) was detected within the fibres in aggregates when visualised by confocal microscopy. Biotinylated F(ab) was rapidly eluted from the patch (97±5% released within 3h) without loss of antigen-binding activity. Patches applied to oral epithelium models successfully delivered the F(ab), with fluorescent F(ab) observed within the tissue and 5.1±1.5% cumulative transepithelial permeation reached after 9h. Neutralising anti-TNFα F(ab) fragments were generated from whole IgG by papain cleavage, as confirmed by SDS-PAGE, then incorporated into patches. F(ab)-containing patches had TNFα neutralising activity, as shown by the suppression of TNFα-mediated CXCL8 release from oral keratinocytes cultured as monolayers. Patches were applied to lipopolysaccharide-stimulated immune-competent oral mucosal ulcer equivalents that contained primary macrophages. Anti-TNFα patch treatment led to reduced levels of active TNFα along with a reduction in the levels of disease-implicated T-cell chemokines (CCL3, CCL5, and CXCL10) to baseline concentrations. This is the first report of an effective device for the delivery of antibody-based biologics to the oral mucosa, enabling the future development of new therapeutic strategies to treat painful conditions.

A Case on Oral Submucous Fibrosis (OSMF)

Oral sub mucous fibrosis (OSMF) is a chronic, insidious, disabling disease of oral mucosa, oropharynx, and rarely, the larynx affecting millions of people worldwide with a prevalence rate of 0.4% in Indian rural population. It results in difficulty in the mouth opening and can be disabling to the patient. It needs a timely diagnosis and thus appropriate treatment can be administered depending on the degree of involvement. A case of oral sub mucous fibrosis occurring in a 55 year old male patient is presented in this article.

ON THE ISSUE OF CLINICAL CLASSIFICATION OF PRECANCEROUS CHANGES IN THE ORAL MUCOSA AND THE LIPS

Diseases of the oral mucosa are an important and complex problem of dentistry. Due to the fact that diseases and pathological conditions of the oral cavity mucosa are diverse in etiology, pathogenesis, clinical manifestations, possible atypical course, have similarity of morphological elements, overall they characterized diseases of various organs and different body systems. In this regard, different methodological approaches are used for creating an optimal classification. P.T. Maksimenko (2012) indicated that each subsequent classification is statistical and intends to take into account diseases and pathological conditions.
 The existing classifications of the oral mucosa diseases have different principles of design. Diseases and lesions of the oral mucosa according to pathogenesis are divided into primary (own), secondary (symptomatic) and syndromes. In all classifications, one of the subset is precancerous changes.
 Nowadays in the world science literature sources there is a term "potentially malignant disorders of the oral cavity" (OPMDs). By this definition, it is recommended to replace the usual terms of precancer, epithelial precursor lesions, premalignant, precancerous, intraepithelial lesions. The definition of potentially malignant oral disorders is included in the WHO classification of Head and Neck Tumors (2017). In March 2020, this classification was supplemented at the WHO International Seminar on the Nomenclature and classification for Oral Cancer and included oral lichenoid lesions, chronic hyperplastic candidiasis and exophytic verrucous oral hyperplasia.
 In 2008, Maksimenko P.T. and co-authors created a working nomenclature and clinical classification of diseases, changes of the oral mucosa, lips and tongue. Symptoms and complexes symptom of the oral mucosa and lips, which are determined by visualization, in a separate section is mentioned the precancerous changes.
 Analysis of the 10-year work of the center for the diagnosis and treatment of diseases of the oral mucosa and the lips, organized at the Department of Dentists Postgraduate Education, made it possible to create some additions to this classification, taking into account atypically occurring precancerous changes and their malignancy. At the same time, regardless of changes in obligate and optional changes in clinical supervision, clinical signs of malignancy should be taken into account:
 
 static course of disease,
 progression of the lesion,
 increasing in the area of the lesion,
 increasing keratinization processes,
 disappearance of boundaries clarity,
 density of the lesion’s base,
 the occurrence of pain.
 
 As well as cytological and histological examination is crucial.
 Conclusion. According to the methodological principles of classifications, data’s of scientific research, information about previously unknown diseases and their manifestation in the oral cavity, it is possible to make additions to existing classifications, as well as to use international ones.
 Future perspectives. Clinical classification of precancerous changes in the oral mucosa and lips, and additions made to it made are very useful for clinicists in making a diagnosis, management of cancer, treatment of patients.

İdiopatik kırmızı lezyon: olgu serisi

The first step in the treatment of periodontal diseases is to mechanically remove plaque and dental calculus, which play a role in the etiology of periodontal diseases, and to establish oral hygiene. In some cases of oral diseases, some drugs and chemical agents can be used for various reasons in addition to mechanical treatment. Corticosteroids are frequently used in oral mucocutaneous diseases because of their strong anti-inflammatory, antiallergic and immunosuppressive effects in the treatment of many other diseases. Although there are different ways of using corticosteroids, the topical form is often preferred in the treatment of oral lesions associated with oral mucosal diseases. This case series aims to evaluate the effect of topical corticosteroid application on red lesions in the gingiva that do not heal with conventional periodontal treatment.

METHODOLOGY FOR STUDYING INTERACTIONS BETWEEN THE MICROBIOCENOSIS OF THE HUMAN ORAL CAVITY AND WHOLE ORGANISM

Stable oral microbiom is associated with the occurrence of dental caries and diseases of periodontal tissues and oral mucosa. Of all the species, streptococci, lactobacilli, actinomycetes and fungi of the genus Candida are found as demonstrating the highest activity. The pathogenicity of a microbial agent may depend on its biological properties. It can also be regarded as an integrated result of the action between several symbionts within the biocenosis or depends on the interaction between the symbionts of the oral biocenosis and the host organism, and this aspect is highlighted in this article. The aim of this study is to apply data from the analysis of the relationship between the host organism and microorganisms, which are known as cariogenic factors, for applying in medical, research and doctor training practice. We analyze the relationship between the host organism and gariogenic microorganisms based on the data obtained from studies of young people with different DMF indices, conducted at the Poltava State Medical University. The paper considers the relationship between the host organism and the oral microbiota in different states of dental caries. The analysis mainly focused on lactobacilli, γ-hemolytic streptococci (which include S. mutans), fungi of the genus Candida, and actinomycetes. The study showed that the number of people with γ-hemolytic streptococci is one hundred percent. This dependence can be expressed by the formula y=100. The number of people with lactobacilli varies from 43% with DMF=0 and over 70% for DMF≥6. This dependence can be expressed by the formula y=13.5x+29. These dependencies change according to the linear laws. The number of people, carriers of actinomycetes, by DMF=0 is also equal to zero. But with an increase in the intensity of caries, the percentage of carriers of these representatives of microflora does not increase according to the linear laws. The formula for such changes is: y=12.5x2-32.5x+20. The percentage of carriers of Candida fungi changes according to the formula: y=5x2-10x+5. These dependences change not according to the linear, but according to the square laws. That is, such changes have a critical point, so-called the bifurcation point. From this point, the relationship between an individual and the indicated representatives of the oral microbiocenosis becomes unstable. Uncertainty arises regarding their further interaction: the "organism-microbiocenosis" system. It can become chaotic or reach a new level of functioning. This research indicates that the relationship between human organisms caring the above mentioned pathogens and representatives of their oral microbiocenoses change differently depending on the state of the oral cavity. One of the indicators of this state is the DMF index. The study of the relationship between microbiocenoses and their carriers should be applied in further scientific work, as well as in teaching activities in teaching sections related to dental caries and periodontal disease.

Arresting the pain syndrome after biopsy collection in oral mucosal diseases

Relevance. Diagnosis of oral mucosa diseases is very difficult. Heterogeneity of the oral mucosa disease manifestation often requires invasive diagnostic methods, which cause the pain syndrome. Timely and complete pain syndrome relief and the impact on all phases of the w ound healing process allow faster patient rehabilitation. The study aimed to examine the effect of Ketanov MD and Cifran CT on the wound process and the pain syndrome intensity after incisional and excisional biopsies to verify the oral mucosa pathology. Materials and methods. The study surveyed 30 people with oral mucosal diseases. The patients (30 subjects) formed two groups: excisional (10 people) and incisional biopsy (20 people). In these groups, we clinically evaluated the course of the wound process, the pain syndrome intensity, oedema phenomena, hyperemia, and the exudate presence. We analysed the quality of life of such patients using a validated Russian v ersion of the questionnaire.Results. On the 3rd day after the biopsy, on top of the generally accepted treatment of oral mucosal diseases and Ketorol MD and Cifran CT intake, the patients noted moderate aching and discomfort when talking and eating. After 14 days, all patients showed an improvement in qualitative and quantitative parameters, the absence of pain and the development of reinfection on therapy with non-steroidal anti-inf lammatory and antibacterial drugs.Conclusion. These drugs have a positive effect on the course of the phases of the wound process. They help reduce the pain response and contribute to faster patient rehabilitation af ter the biopsies in oral mucosal diseases.

Optimization of examination methods in the diagnosis of precancerous and malignant oral mucosal lesions

Relevance. The prevalence of oral mucosal diseases among the Russian population varies from 3% to 20% [7]. Precancerous and malignant diseases are a particular problem. More than nine thousand new cases of malignant oral mucosal lesions are registered annually in the Russian Federation, and yearly mortality rates reach 34%. Unfortunately, despite being externally located, the rate of late-diagnosed malignant oral mucosal neoplasms reaches 60%-70% in various regions of the Russian Federation [2]. Thus, one of the most urgent problems in dentistry and oncology is the early diagnosis of precancerous and malignant oral mucosal lesions. As a rule, precancerous lesions are not diagnosed at an early stage since there are no visible clinical signs, and therefore patients do not seek medical attention [8]. An accurate diagnosis requires both basic and additional examination methods. In turn, the conventional methods include traditional inspection, which largely depends on the clinician's experience. Besides the clinical examination, additional techniques are used, namely, fluorescence examination and biopsy [9-11]. Subsequently, optimizing the diagnosis of oral mucosal lesions is the most promising direction in practical healthcare, both in the dental and oncology practice, for early detection and reduction of advanced stages of malignant oral mucosal lesions. Thus, the study confirmed that the development and application of modern approaches are necessary for the early diagnosis of oral mucosal lesions. Purpose. The study aimed to improve the outcome of oral mucosal precancerous and malignant lesion diagnosis by upgrading examination methods. Material and methods. The study included 147 patients with oral mucosal lesions, referred to the oncologists of Samara regional clinical oncology centre by the city polyclinics. The patients were divided into two groups according to the examination methods. The control group patients, 63 people, had a conventional examination by a dentist (patient interview, inspection, palpation) and an incisional biopsy by an oncologist. The main group consisted of 84 patients, who, besides conventional dental examination, were evaluated by a new – developed and put into practice – technique with point and index score and subsequent incisional biopsy performed by an oncologist. The studied patients were comparable by gender, age and localization. The study assessed the effectiveness of the new method for the diagnosis of precancerous lesions (PL) and malignant lesions (ML) by matching the Need for Histology Verification Index (NHVI) value equal to 5 points or more and histopathology results.Results. In the main group, 71 out of 84 patients scored 5 or more according to the new method, and 13 patients scored less than 5. Patients with a low score had a non-surgical treatment, 11 patients reached remission, and two patients were referred to an oncologist for a biopsy, which confirmed oral mucosa PL and ML. The patient complaints in both groups demonstrated that pain and bleeding were more frequent in the control group compared to the main one. The evaluation of clinical examination data revealed more erosions in the control group and nonremovable plaque and hyperplasia in the main group. The incisional biopsy detected more PL and ML in the main group (p = 0.001), and early malignant lesions were in 23% versus 5% in the control group. The new method specificity in oral mucosal PL and ML diagnosis was 55%, sensitivity – 97%, accuracy – 87%, whereas the conventional examination specificity was 28%, sensitivity – 84%, and accuracy – 60%.Conclusion. The administered improved method for examination of patients with oral mucosal lesions and the compulsory use of autofluorescence examination and risk factor score assessment allowed us to identify PL and ML in 88% and to diagnose more PL and ML at early stages, which explains the need to use this method in the clinical practice for the early diagnosis of lesions.

Characteristics analysis of oral clinical trials registered in Chinese Clinical Trial Registry

Objective: To investigate the project characteristics of oral clinical trials registered in Chinese Clinical Trial Registry (ChiCTR), and to provide reference for medical institutions to improve the quality of oral clinical trials and formulate management systems. Methods: The ChiCTR database was retrieved to collect all the oral-related clinical trials from the time of database establishment to July 25, 2021. Those clinical trials were analyzed statistically in respect of name of registered project, registration time, registration status, regional distribution of research institutions, approval status by ethics committee, sample size, source of funds, involved disease, research type and design, randomization method, and whether blind method was adopted. Results: A total of 778 oral clinical trials, which studied mainly in the oral and maxillofacial diseases, periodontal tissue diseases, oral implant diseases, oral mucosal diseases, and oral prosthetic diseases, were retrieved in the database. Beijing, Sichuan, Shanghai, Guangdong and Hubei were major regions where oral clinical trials were carried out, accounting for 69.68% (772/1 108) of the total. The top four funding sources were hospital finance [24.93% (186/746)], local government finance [22.39% (167/746)], self-financing [17.69% (132/746)], and national finance [12.47% (93/746)]. For the types of researches, 520 interventional studies and 244 observational studies were identified (accounting for 66.84% and 31.36%, respectively). The research designs were dominated by ways of randomized control (381, 48.97%), of which 240 (62.99%) trials were with missing or unspecified blinding methods. Conclusions: Oral clinical trials are increasing year by year, but they are regionally imbalanced, and still need to be further improved in registration information and research design. Administrative departments should pay more attention to strengthen the publicity and education on the registration and publication mechanism of clinical trials, and enhance researchers' cognitions in clinical trials registration and clinical trials design.

Role of human papillomavirus infection in the development of oral leukoplakia and oral squamous cell carcinoma

BACKGROUND: The annual global oral cancer morbidity is more than 650,000 cases. Between 2007 and 2017 in the Russian Federation, the absolute number of first-ever diagnosed oral cavity malignant neoplasms is constantly increasing. According to many authors, oral squamous cell carcinoma is associated with pre-existing leukoplakia. This disease is the most common and prone to malignant precancerous oral mucosa disease. Human papillomaviruses (HPV) are crucial players in this process. However, the relationship between HPV infection in leukoplakia and oral squamous cell carcinoma remains unclear.
 AIM: To establish the HPV role in the development of leukoplakia and squamous cell carcinoma of the oral mucosa.
 MATERIAL AND METHODS: The study included 39 patients with verified various forms of oral leukoplakia and oral squamous cell carcinomas. The squamous cell carcinoma group included 19 patients. The oral leukoplakia group is consisted of 20 patients. The control group of healthy volunteers is represented by 22 patients with clinically healthy oral mucosa. HPV deoxyribonucleic acid (DNA) was detected and quantified in clinical materials by polymerase chain reaction with hybridization-fluorescence detection AmpliSense HPV HRS screen-titer-FL for HPV types 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, and 68 and AmpliSense HPV 6/11-FL for HPV types 6 and 11.
 RESULTS: HPV types 6 and 11 DNA were isolated in three cases with insignificant clinical disease from 19 samples taken from patients with oral squamous cell carcinoma. In groups of leukoplakia and healthy volunteers, no high oncological risk was identified. Despite a HPV-positive expression in squamous cell carcinoma samples is detected, no significant difference was found between the squamous cell carcinoma group and control group (p=0.09).
 CONCLUSION: No significant association was noted between HPV and oral squamous cell carcinoma; however, a low carcinogenic-risk HPV was detected in 15.7% of carcinoma samples. No HPV contamination was detected in patients with leukoplakia and healthy groups.

A Prospective Comparative Analysis Regarding the Assessment of Oral Mucosal Disease Using a Validated Questionnaire to Improve the Teaching of Dental Students.

Structured examination and treatment are essential in medicine. For dental students, a structured approach to the assessment of oral mucosal lesions is missing thus far. To validate an approach, a structured questionnaire was compared with the habitually used free description of oral lesions (white lesions, ulcers, hyperplasia). Thirty-three dental students were divided into two groups (Group 1 (n = 17) used the free description; Group 2 (n = 16) used a guided questionnaire) to characterize mucosal lesions in patients and make a tentative diagnosis. Although no difference was found between the groups regarding the suspected diagnosis or the histopathological findings, there was a significant advantage of the structured questionnaire in all aspects of the description compared to the free description (p = 0.000018). Thus, a structured description is an important aspect in the evaluation of oral mucosal changes, and a guided questionnaire should be implemented in the study of dentistry.

Composite dissolvable microneedle patch for therapy of oral mucosal diseases.

A composite microneedle patch (MN patch) is developed for oral transmucosal administration. To improve the oral transmucosal drug delivery efficiency, the composite MN patch is designed to consist of an array of 100 dissolvable microneedles (MNs) with drug-loaded tips and a backing layer. The MNs are composed of two parts, the hyaluronic acid (HA) tip part and the polyvinylpyrrolidone (PVP) base part. Due to the small size and sufficient mechanical strength, the HA-PVP MNs can painlessly penetrate the oral mucosa barrier and deliver drugs directly to the basal layer or submucosa. Betamethasone sodium phosphate (BSP), as the model drug, is concentrated in the HA tip parts to avoid the drug waste caused by mucosa elasticity. Considering the special moist environment and saliva flow in the mouth, a double-layer backing layer composed of a poly(vinyl alcohol) (PVA) adhesive layer and an ethyl cellulose (EC) waterproof layer is designed and constructed, which could reduce the saliva flow effects. The in vitro and in vivo results demonstrate that the MN patch could achieve rapid and efficient BSP release in oral mucosa due to the rapid dissolution of HA. The proposed MN patch provides a novel strategy for the therapy of oral mucosal diseases.

Diagnosis and management of oral mucosal conditions commonly seen in the elderly population

##Learning outcomes recognise the older adult as a unique subset of patient; develop familiarity with the appearance and aetiology of various oral mucosal conditions that can present in the older adult; and, develop awareness of common treatment modalities for oral mucosal diseases.

Function of normal oral mucosa revealed by single-cell RNA sequencing.

The functions of oral mucosa include barrier, sensation, and secretion. The barrier protection function is particularly important, which includes physical barrier and immunological barrier. Few studies have revealed the function of oral mucosa by displaying the map of normal oral mucosal cells from the perspective of single cells. Here, single-cell transcriptome sequencing was used to bring a relatively comprehensive map of the normal oral mucosal cells. In total,26,398 cells from three cases of normal oral mucosa were analyzed by single-cell RNA-sequencing and 14 distinct cell groups were defined, 7 of which were immune cells. We performed subgroup classification and heterogeneity analysis of epithelial cells, T cells, and macrophagocytes, which found a subpopulation of epithelial cells with high expression of major histocompatibility complex class II molecules, a subpopulation CD8+ GZMK+ T cells, and two kinds of active macrophagocytes. Meanwhile, we identified ligand-receptor pairs among the major cell types to explore the interactions and how they maintain the homeostasis of normal oral mucosa. Based on these results, the epithelial barrier function, immunological barrier function, and potential maintenance function of stromal cells in the oral mucosa were described at the single-cell level, which provides basic data resources for further studies of oral mucosal diseases.

The Evaluation of Healing Properties of Galium verum-Based Oral Gel in Aphthous Stomatitis in Rats.

Although oral ulcers represent one of the most frequent oral mucosal diseases, the available treatment is not sufficient to provide complete ulcer recovery without side-effects. Therefore, the aim of our study was to prepare a mucoadhesive oral gel based on Galium verum ethanol extract (GVL gel) and reveal its healing effects in the model of aphthous stomatitis in rats. Rats with oral ulcers were divided into the following groups: control (untreated), gel base (ulcer was treated with the gel base, three times per day for 10 days), and GVL gel group (the ulcer was treated with GVL gel in the same way as the gel base). Animals from each group were sacrificed on days 0, 3, 6, and 10 for collecting blood and ulcer tissue samples. Healing properties of oral gel were determined by clinical evaluation, as well as biochemical and histopathological examinations. Our findings suggest a significant decrease in the ulcer size in GVL gel group, with healing effects achieved through the alleviation of oxidative stress, reduction in COX-2 immunopositivity, and increase in collagen content in buccal tissue. Significant ulcer repairing potential of GVL gel highlights this oral mucoadhesive gel as a promising tool for prevention and treatment of RAS.

Janus mucosal dressing with a tough and adhesive hydrogel based on synergistic effects of gelatin, polydopamine, and nano-clay

There are many problems and challenges related to the treatment of highly prevalent oral mucosal diseases and oral drug delivery because of a large amount of saliva present in the oral cavity, the accompanying oral movements, and unconscious swallowing in the mouth. Therefore, an ideal oral dressing should possess stable adhesion and superior tough strength in the oral cavity. However, this fundamental requirement greatly limits the use of synthetic adhesive dressings for oral dressings. Here, we developed a mussel-inspired Janus gelatin-polydopamine-nano-clay (GPC) hydrogel with controlled adhesion and toughness through the synergistic physical and chemical interaction of gelatin (Gel), nano-clay, and dopamine (DA). The hydrogel not only exhibits strong wet adhesion force (63 kPa) but also has high toughness (1026 ± 100 J m−3). Interfacial adhesion of hydrogels is achieved by modulating the interaction of catechol groups of the hydrogel with specific functional groups (e.g., NH2, SH, OH, and COOH) on the tissue surface. The matrix dissipation of the hydrogel is regulated by physical crosslinking of gelatin, chemical crosslinking of gelatin with polydopamine (Michael addition and Schiff base formation), and nano-clay-induced constraint of the molecular chain. In addition, the GPC hydrogel shows high cell affinity and favors cell adhesion and proliferation. The hydrogel's instant and strong mucoadhesive properties provide a long-lasting therapeutic effect of the drug, thereby enhancing the healing of oral ulcers. Therefore, mussel-inspired wet-adhesion Janus GPC hydrogels can be used as a platform for mucosal dressing and drug delivery systems. Statement of significanceIt is a great challenge to treat oral mucosal diseases due to the large amount of saliva present in the oral cavity, the accompanying oral movements, unconscious swallowing, and flushing of drugs in the mouth. To overcome the significant limitations of clinical bioadhesives, such as weakness, toxicity, and poor usage, in the present study, we developed a simple method through the synergistic effects of gelatin, polydopamine, and nano-clay to prepare an optimal mucosal dressing (Janus GPC) that integrates Janus, adhesion, toughness, and drug release property. It fits effectively in the mouth, resists saliva flushing and oral movements, provides oral drug delivery, and reduces patient discomfort. The Janus GPC adhesive hydrogels have great commercial potential to support further the development of innovative therapies for oral mucosal diseases.

Intravital longitudinal cellular visualization of oral mucosa in a murine model based on rotatory side-view confocal endomicroscopy.

Oral mucosa is a soft tissue lining the inside of the mouth, protecting the oral cavity from microbiological insults. The mucosal immune system is composed of diverse types of cells that defend against a wide range of pathogens. The pathophysiology of various oral mucosal diseases has been studied mostly by ex vivo histological analysis of harvested specimens. However, to analyze dynamic cellular processes in the oral mucosa, longitudinal in vivo observation of the oral mucosa in a single mouse during pathogenesis is a highly desirable and efficient approach. Herein, by utilizing micro GRIN lens-based rotatory side-view confocal endomicroscopy, we demonstrated non-invasive longitudinal cellular-level in vivo imaging of the oral mucosa, visualizing fluorescently labeled cells including various immune cells, pericytes, nerve cells, and lymphatic and vascular endothelial cells. With rotational and sliding movement of the side-view endomicroscope on the oral mucosa, we successfully achieved a multi-color wide-area cellular-level visualization in a noninvasive manner. By using a transgenic mouse expressing photoconvertible protein, Kaede, we achieved longitudinal repetitive imaging of the same microscopic area in the buccal mucosa of a single mouse for up to 10 days. Finally, we performed longitudinal intravital visualization of the oral mucosa in a DNFB-derived oral contact allergy mouse model, which revealed highly dynamic spatiotemporal changes of CSF1R or LysM expressing immune cells such as monocytes, macrophages, and granulocytes in response to allergic challenge for one week. This technique can be a useful tool to investigate the complex pathophysiology of oral mucosal diseases.

Engineering a 3D In Vitro Model of Human Gingival Tissue Equivalent with Genipin/Cytochalasin D.

Although three-dimensional (3D) co-culture of gingival keratinocytes and fibroblasts-populated collagen gel can mimic 3D structure of in vivo tissue, the uncontrolled contraction of collagen gel restricts its application in clinical and experimental practices. We here established a stable 3D gingival tissue equivalent (GTE) using hTERT-immortalized gingival fibroblasts (hGFBs)-populated collagen gel directly crosslinked with genipin/cytochalasin D and seeding hTERT-immortalized gingival keratinocytes (TIGKs) on the upper surface for a 2-week air–liquid interface co-culture. MTT assay was used to measure the cell viability of GTEs. GTE size was monitored following culture period, and the contraction was analyzed. Immunohistochemical assay was used to analyze GTE structure. qRT-PCR was conducted to examine the mRNA expression of keratinocyte-specific genes. Fifty µM genipin (G50) or combination (G + C) of G50 and 100 nM cytochalasin D significantly inhibited GTE contraction. Additionally, a higher cell viability appeared in GTEs crosslinked with G50 or G + C. GTEs crosslinked with genipin/cytochalasin D showed a distinct multilayered stratified epithelium that expressed keratinocyte-specific genes similar to native gingiva. Collagen directly crosslinked with G50 or G + C significantly reduced GTE contraction without damaging the epithelium. In summary, the TIGKs and hGFBs can successfully form organotypic multilayered cultures, which can be a valuable tool in the research regarding periodontal disease as well as oral mucosa disease. We conclude that genipin is a promising crosslinker with the ability to reduce collagen contraction while maintaining normal cell function in collagen-based oral tissue engineering.

Comparison of Effects of Honey and 0.1% Triamcinolone Acetonide in the Management of Recurrent Aphthous Stomatitis - A Randomised, Single-Blind Study

Introduction: Recurrent aphthous stomatitis (RAS) is the most common oral mucosal disease known to humans and are characterized by the development of painful, recurring solitary or multiple ulcers confined to the oral mucosa. RAS is often painful and worsens during drinking and talking. Aim: To evaluate and compare the effects of honey and 0.1% triamcinolone acetonide in patients with ulcers of recurrent aphthous stomatitis minor. Materials and Methods: Sixty patients having RAS minor were divided into two groups of 30 participants each by simple random sampling. Group A was treated with application of 0.5 ml of honey thrice a day for a week or until complete healing of the ulcer. Group B was treated with application of 5 g of 0.1% triamcinolone acetonide oral paste thrice a day for a week or until complete healing of the ulcer. For all the individuals, baseline parameters like the size of the ulcer, pain, and burning sensation were recorded on day 0, reassessment was done on day 4 and day 7, followed by statistical analysis. Results: The study showed that the reduction in the size of ulcers, VAS score for pain, and VAS score for burning sensation were similar in patients receiving honey as compared to patients receiving 0.1% triamcinolone acetonide. No adverse effects were reported in patients receiving honey, and there were mildly adverse reactions in some patients receiving 0.1% triamcinolone acetonide oral paste. Conclusion: Honey, a natural anti-inflammatory agent, seems to hold immense potential as an alternative medication for managing RAS minor ulcers. It has better patient compliance as compared to 0.1% triamcinolone acetonide.

Oral Health Impact Profile in adult Polish population.

Oral Health Related Quality of Life (OHRQoL) is a significant patient-oriented outcome which should be considered in diagnostic, decision making and therapeutic process by dentists. To evaluate the influence of a condition of teeth, oral mucosa and dentures on the OHRQoL among patients seeking dental treatment in the University Dental Clinic (UDC) in Kraków, Poland. 250 patients took part in a cross-sectional study conducted in the UDC in Kraków. Collected clinical data included: oral mucosa and periodontal condition, number of Decayed, Missed and Filled Teeth (DMFT index), presence of dentures. Questionnaire part of the study has been based on OHIP-14 questionnaire. The condition of teeth worsened OHRQoL in following groups of patients: first-time visitors, smokers, with periodontal diseases and without oral mucosal diseases. There was a positive significant correlation between number of decayed teeth and OHRQoL related to teeth. The condition of oral mucosa significantly impaired OHRQoL in: women, patients with oral mucosal diseases and those who undergone prosthetic treatment in the last year. There was a correlation between number of missing teeth, DMFT index, age and subscale 2 of the form. The condition of dentures significantly impaired the OHRQoL in patients: who used removable dental prostheses, undergone prosthetic treatment in the last year and females. There was a positive correlation between subscale 3 of the questionnaire and number of missing teeth and DMFT index and a negative correlation with number of filled teeth within this subscale. Evaluation of patient's OHRQoL is one of important components for successful dental treatment.