Periodontal treatment and microbiome-targeted therapy in management of periodontitis-related nonalcoholic fatty liver disease with oral and gut dysbiosis

A growing body of evidence from multiple areas proposes that periodontal disease,accompanied by oral inflammation and pathological changes in the microbiome,induces gut dysbiosis and is involved in the pathogenesis of nonalcoholic fatty liver disease(NAFLD).A subgroup of NAFLD patients have a severely progressive form,namely nonalcoholic steatohepatitis(NASH),which is characterized by histological findings that include inflammatory cell infiltration and fibrosis.NASH has a high risk of further progression to cirrhosis and hepatocellular carcinoma.The oral microbiota may serve as an endogenous reservoir for gut microbiota,and transport of oral bacteria through the gastro-intestinal tract can set up a gut microbiome dysbiosis.Gut dysbiosis increases the production of potential hepatotoxins,including lipopolysaccharide,ethanol,and other volatile organic compounds such as acetone,phenol and cyclopentane.Moreover,gut dysbiosis increases intestinal permeability by disrupting tight junctions in the intestinal wall,leading to enhanced translocation of these hepatotoxins and enteric bacteria into the liver through the portal circulation.In particular,many animal studies support that oral administration of Porphyromonas gingivalis,a typical periodontopathic bacterium,induces disturbances in glycolipid metabolism and inflammation in the liver with gut dysbiosis.NAFLD,also known as the hepatic phenotype of metabolic syndrome,is strongly associated with metabolic complications,such as obesity and diabetes.Periodontal disease also has a bidirectional relationship with metabolic syndrome,and both diseases may induce oral and gut microbiome dysbiosis with insulin resistance and systemic chronic inflammation cooperatively.In this review,we will describe the link between periodontal disease and NAFLD with a focus on basic,epidemiological,and clinical studies,and discuss potential mechanisms linking the two diseases and possible therapeutic approaches focused on the microbiome.In conclusion,it is presumed that the pathogenesis of NAFLD involves a complex crosstalk between periodontal disease,gut microbiota,and metabolic syndrome.Thus,the conventional periodontal treatment and novel microbiome-targeted therapies that include probiotics,prebiotics and bacteriocins would hold great promise for preventing the onset and progression of NAFLD and subsequent complications in patients with periodontal disease.

Combination of Diclofenac Potassium and Propolis in the Therapy of Oral Aphthosis: A Randomized, Clinical, Double-Blind, Placebo-Controlled Study

Background: Oral aphthosis has a negative impact on oral health. This study aimed to assess the effectiveness of gel formulation including diclofenac and propolis in the treatment of oral ulcers. Methods: Participants included 100 normal individuals with aphthous, excluding those having allergies to any ingredient in the diclofenac formulation. Patients were randomly assigned into two groups: one group received treatment with a combination of diclofenac potassium 3% (10 mg/g, 60 g) and propolis 5% gel (Group II), and the other group received a placebo (Group 1). The patient was evaluated using standard digital photographs and chronic oral mucosal questionnaires on days 1, 3, 5, and 7 after healing. Utilizing the Mann-Whitney U test, the two groups were compared. Each group’s changes were examined using Friedman’s test. Results: There was a statistically dramatic change over time in Group II. After one day, the median total score dropped statistically significantly, and from one to three days with effect size (d) 2.485, Group II demonstrated 48% complete healing and 52% partial healing, while Group I demonstrated 4% partial healing and 96% no change. Effect size (V): 0.995. Conclusions: The combination of diclofenac and propolis provided instant relief and an affordable new regimen for treating oral aphthosis.

Development of a mouse model of arecoline-induced oral mucosal fibrosis

Objective: To develop a BALB/c mouse model of oral submucous fibrosis(OSF)induced by arecoline and to exhibit an accumulation of collagen and angiogenesis changes.Methods: BALB/c mice were randomly assigned to either the control(distilled water) or experimental group(arecoline)(n = 40). Eight mice from each group were sacrificed every 4 weeks since 8 weeks post treatment. Changes in histopathologic features, levels of collagen type Ⅰ and collagen type Ⅲ, and angiogenesis were measured.Results: In the 8th week, epithelium atrophy, collagen cumulation and micrangium pathologic changes in the lamina propria were observed in the oral mucosa. In the 20th week, hyaline degeneration of the connective tissues was observed on the tongue and palate mucosa. The angiogenesis and collagen type Ⅰ changed significantly as the diseases advanced(P < 0.05); however, collagen type Ⅲ was not statistically different.Conclusions: An OSF model involving mice can be rapidly induced by drinking a highdose of arecoline. OSF angiogenic changes in mice primarily decrease and collagen accumulation is mainly collagen type Ⅰ.

Glycolysis reprogramming in cancer-associated fibroblasts promotes the growth of oral cancer through the lncRNA H19/miR-675-5p/PFKFB3 signaling pathway

As an important component of the tumor microenvironment,cancer-associated fibroblasts(CAFs)secrete energy metabolites to supply energy for tumor progression.Abnormal regulation of long noncoding RNAs(lncRNAs)is thought to contribute to glucose metabolism,but the role of lncRNAs in glycolysis in oral CAFs has not been systematically examined.In the present study,by using RNA sequencing and bioinformatics analysis,we analyzed the lncRNA/mRNA profiles of normal fibroblasts(NFs)derived from normal tissues and CAFs derived from patients with oral squamous cell carcinoma(OSCC).LncRNA H19 was identified as a key lncRNA in oral CAFs and was synchronously upregulated in both oral cancer cell lines and CAFs.Using small interfering RNA(siRNA)strategies,we determined that lncRNA H19 knockdown affected proliferation,migration,and glycolysis in oral CAFs.We found that knockdown of lncRNA H19 by siRNA suppressed the MAPK signaling pathway,6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 3(PFKFB3)and miR-675-5p.Furthermore,the lncRNA H19/miR-675-5p/PFKFB3 axis was involved in promoting the glycolysis pathway in oral CAFs,as demonstrated by a luciferase reporter system assay and treatment with a miRNA-specific inhibitor.Our study presents a new way to understand glucose metabolism in oral CAFs,theoretically providing a novel biomarker for OSCC molecular diagnosis and a new target for antitumor therapy.

Pyoderma Gangrenosum with Oral Involvement–Case Report and Review of the Literature

Pyoderma gangrenosum （PG） is a rare, noninfectious neutrophilic dermatosis. Clinically, it begins with sterile pustules that rapidly progress into painful ulcers of variable depth and size with undermined violaceous borders. The diagnosis of PG is based on the history of an underlying disease, a typical clinical presentation, histopathology, and exclusion of other diseases. The peak incidence occurs between the ages of 20 to 50 years with women being more often affected than men. There have been very few reports of pyoderma gangrenosum with oral mucosal involvement. Oral lesions in previously reported cases have included ulcers of varying sizes from a few mm to several cm and have been reported to have been found on the tongue, soft and hard palate, buccal mucosa, and gingiva. Some of these oral lesions have been associated with ulcerative colitis, inflammatory bowel disease, and polycythemia rubra vera. A few cases were reported with biopsy findings, the histological picture being nonspecific, showing ulceration, and necrosis with inflam- matory cell infiltrate. A peculiar case of pyoderma gan- grenosum with an oral lesion is presented here, and the differential diagnosis is discussed.

Quality of life and oral potentially malignant disorders: Critical appraisal and prospects

Quality of life(QoL) is a vital and often required health outcome measure that is relevant to patient care. A healthy oral cavity enables person to perform daily activities without any limitations. However, any disturbance may result in impaired QoL. The oral health-remains an essential element of people's health and well-being. In recent years, the tradition of clinical practice and research has been changed by incorporating QoL assessment, as it helps in assessment of patients' needs and monitoring treatment responses. Oral potentially malignant disorders(OPMDs) are a group of chronic disorders including oral leukoplakia(OL), oral lichen planus and oral submucous fibrosis(OSF). It is evident that patients with OPMDs experience significant health-related symptoms, functional limitations and psycho-social impairment, compromising their Qo L. Moreover, the worsening of Qo L has been associated with advanced stages of OPMDs. Despite of increasing number of OPMD cases in recent decades, limited literature is available regarding QoL in this population. Although, there is higher prevalence of habit-related OPMDs, particularly OSF and OL in Southern Asian countries, only a few studies have been performed in these populations. Moreover, these studies administered generic Qo L instruments, which offer less sensitivity to clinical changes. However, condition-specific instruments are more sensitive and allows better measurement of QoL. As the impacts of different conditions on OHRQoL may vary, the development and validation of a QoL instrument specific to each clinical entity of OPMDs is currently needed.

Photodynamic therapy of oral leukoplakia and orallichen planus using methylene blue:A pilot study

Premalignant lesions like oral lichen planus(OLP),oral leukoplakia(OL)has a fair probability of transforming into malignancy and they are perverse toward conventional therapies.Photody-namic therapy(PDT)has been considered as an alternative/complimentary therapeutic mo-dality for the management of premalignant lesions.In this study,methylene blue mediated photodynamic therapy(MB-PDT)was used as a possible alternative method for the treatment of OLP and OL.A total of 15 OLP lesions and 13 OL lesions were enrolled in the study.The patients were irradiated 1using metal halide lamp filtered at 630土10nm,with a light exposure dose of 120 J/cm^(2) per stting.For the OLP lesions,MB-PDT was performed once a week for four weeks and for the OL lesions,MB-PDT was performed twice a week for three weeks.Lesions were evaluated pre-and post-and at follow-u sessions by changes in sign and symptom scores,and size of lesions.We have observed a 53.3%of complete reduction in the treated OLP lesions and their decrease in size,sign and symptom score after treatment and at follow-up session was statistially signifcant.We have also observed complete response for one OL lesion of the 13 treated lesions.The result indicates that MB-PDT is an effective modality in management of OLP and OL.Among the two types of premalignancies treated with MB-PDT,OLP lesions responded much better than that of OL.

Fluconazole Susceptibility and Genotypic Heterogeneity of Oral Candida albicans Colonies from the Patients with Cancer Receiving Chemotherapy in China

Aim To identify heterogeneity of Candida albicans （C. albicans） isolated from the population with cancer in China by using identification medium, subculture molecular typing, and antifungal susceptibility test. Methodology Oral cheek mucosal specimens from 52 cancer patients receiving chemotherapy were cultured on CHROMagar CandidaTM plates for Candida identification. All the C. albicans colonies on the plates were subcultured and reconfirmed by API20C, then submitted to the antifimgal drug susceptibility test with fluconazole and molecular typing using randomly amplified polymorphic DNA-PCR （RAPD） with primers RSD6 and RSD12.Results 54% （28/52） patients were oral yeast carriage in which C. albicans predominated. More than 7 C. albicans colonies were isolated from each of 12 patients （Group A）, while less than 5 colonies were isolated from each of 16 patients （Group B）. RSD6 and RSD12 were successful in eliciting 17 （A1-A17） and 2 （B1-B2） genotypes, respectively from among the 205 isolates. The two primers were combined to generate 21 genotypes. The C. albicans isolates obtained from the same patient and episode showed a diversity for fluconazole revealed by MIC50 and MIC90. Conclusion The heterogeneity of the C. albicans colonies isolated from the same patients can be detected. C. albicans with varied fluconazole susceptibility and genotypic characteristics may coexist in the same oral Candida population.

Role of human papillomavirus in the pathogenesis of oral squamous cell carcinoma

Oral cancer is one of the most common cancers and it constitutes a major health problem particularly in developing countries. Oral squamous cell carcinoma(OSCC) represents the most frequent of all oral neoplasms. Several risk factors have been well characterized to be associated with OSCC with substantial evidences. While tobacco and alcohol are the primary risk factors for OSCC development, many epidemiological studies report a strong association with human papillomavirus(HPV) in a subset of OSCC. This article presents our current knowledge on the relationship between HPV and development of OSCC. HPVs are DNA viruses that specifically target the basal cells of the epithelial mucosa. Most experimental data are consistent with the hypothesis that HPV plays a causal role in oral carcinogenesis. Genotypes, such as HPV1 infect epidermal cells, whereas HPV6, 11, 16 and 18 infect epithelial cells of the oral cavity and other mucosal surfaces. Several studies have shown that there is an increased risk of head and neck cancer in the two major HPV 16 oncogenes E6 and E7-positive patients. The presence of antibodies to HPV E6 and E7 proteins was found to be more associated with tumors of the oro-pharynx than of the oral cavity. However, HPV alone appears to be insufficient as the cause of OSCC but requires other co-factors. Although a viral association within a subset of OSCC has been shown, the molecular and histopathological characteristics of these tumors have yet to be clearly defined.

Formulation of a new phenytoin-containing mucoadhesive and evaluation of its healing effects on oral biopsy ulcers

Background and Objective: Several studies have shown the wound healing effect of topical phenytoin, which is applied by its effect on connective tissue intracellular matrix. However, there are still some controversies about its effect on various kinds of wounds, especially in the experimental models. This study is aimed at evaluating the effect of mucoadhesive paste compared to phenytoin mucoadhesive paste on wound healing after oral biopsy. Material and Methods: In this double blind randomized clinical trial, 20 patients who were eligible for oral biopsy were allocated into the case and control groups. After the biopsy, patients having ulcers ranging between one and two centimeters were treated by simple or 1% phenytoin mucoadhesive paste. All patients were instructed to apply their paste at least three times a day for five days after the biopsy. Patients in both groups were evaluated every other day for size of the ulcer, degree of pain and diameter of the inflammatory halo. Statistical analysis was done using SPSS software and Mann-Whitney test. Results: After the second and third appointments, it was observed that the rate of wound healing and decrease in the size of the ulcers were significantly quicker in the treatment group (p = 0.001 and p = 0.003 respectively) and the patients in the phenytoin group reported less pain. Diameter of the inflammatory halo was not significantly different between two groups. Conclusion: Applying 1% phenytoin mucoadhesive paste on biopsy ulcers resulted in accelerated wound healing and decrease in pain, but had no effect on the diameter of the inflammatory halo.

Mystery behind labial and oral melanotic macules: Clinical, dermoscopic and pathological aspects of Laugier-Hunziker syndrome

Labial and oral melanotic macules are commonly encountered in a broad range of conditions ranging from physiologic pigmentation to a sign of an underlying life-threatening disease. Although Laugier-Hunziker syndrome(LHS) shares some features of labial and oral pigmentation with a variety of conditions, it is a benign and acquired condition, frequently associated with longitudinal melanonychia. Herein, the demographic, clinical, dermoscopic, and pathological aspects of LHS were reviewed comprehensively. The important differential diagnoses of mucocutaneous and nail pigmentation are provided. An accurate diagnosis is crucial to design a reasonable medical strategy, including management options, malignant transformation surveillance, and psychological support. It is important that clinicians conduct long-term follow-up and surveillance due to the potential risks of malignant transformation and local severe complications in some conditions.

Spindle cell lipoma of the mandibular mucogingival junction: a case report of unusual oral neoplasm

Spindle cell lipoma（SCL） is a benign lipomatous neoplasm typically located in the posterior neck and back of older males. It presents as a well-circumscribed mass in the buccal mucosa, tongue, floor of the mouth or hard palate. There are only two case reports of SCL in the gingiva and alveolar ridge. Here, we report a case of SCL in the mandibular mucogingival junction of a 68-year-old male. Clinical,histopathological and immunohistochemical findings are presented. Although oral SCL is rare, it should be considered in the differential diagnosis of spindle cell neoplasms occurring in the oral cavity.

A rare case of intraoral lipoma in a six year-old child:a case report

One type of soft tissue lesions of the oral cavity is lipoma, which is a kind of benign tumor composed of mature lipid cells. Although the lipoma presents as one of the most common mesenchymal neoplasms, most tend to develop on the trunk and proximal portions of the extremities. However, lipomas in the oral and maxillofacial region are much less frequent. Here we present a case of an intraoral lipoma in a six year-old child.

A portable illumination device for photodynamic therapy of the oral cavity

Photodynamic therapy(PDT)is a minimally invasive method for treating oral leukoplakia In this paper,we propose a portable PDT device consisting of a flexible circuit board with a liquid flow cooling module on the back.The light source size was 17 mm×11 mm×4 mm,and the iradiation area of the light source was up to 100mm'.The iradiance range of this device was from 10mW/cm^(2)to 100mW/cm^(2).Simulation and experimental results showed that the irradiance cofficient variation for a treatment area of 81 mm was less than 7%.At an irradiance of 100 mW/cm a device surface temperature of lower than 42℃ can be achieved to satisfy the safety requirements under the con-ditions that the temperature of cooling liquid is 10℃ and the liquid flow speed is above 12 mL/min.

Identification of oral squamous cell carcinoma in optical coherence tomography images based on texture features

Surgical excision is an effective treatment for oral squamous cell carcinoma(OSCC),but exact intraoperative differentiation OSCC from the normal tissue is the first premise.As a noninvasive imaging technique,optical coherence tomography(OCT)has the nearly same resolution as the histopathological examination,whose images contain rich information to assist surgeons to make clinical decisions.We extracted kinds of texture features from OCT images obtained by a home-made swept-source OCT system in this paper,and studied the identification of OSCC based on different combinations of texture features and machine learning classifiers.It was demonstrated that different combinations had different accuracies,among which the combination of texture features,gray level co-occurrence matrix(GLCM),Laws'texture measnres(LM),and center symmetric auto-correlation(CSAC),and SVM as the classifier,had the optimal comprehensive identification effect,whose accuracy was 94.1%.It was proven that it is feasible to distinguish OSCC based on texture features in OCT images,and it has great potential in helping surgeons make rapid and accurate decisions in oral clinical practice.

Intraoperative imaging of oral-maxillofacial lesions using optical coherence tomography

Histopathological examination is still the gold standard for diagnoses of oral-maxillofacial lesions,but it is invasive and time-consuming.Optical coherence tomography(OCT)provides a kind of noninvasive,label-free,real-time and high-resolution imaging technology.In this study,in order to assess the feasibility of OCT in oral clinical application,fresh excised tissue specimens from 59 patients undergoing oral-maxillofacial surgery were imaged in detail by using a benchtop sweptsource OCT system.It is shown that different lesions or tissues can be obviously distinguished based on their different microstructural features in OCT images,and the features are similar to those of their corresponding histopathological images.It is proven that OCT has great feasibility and potential as a diagnostic aid for surgeons in oral medicine.

Inhibition of EGFR Suppresses Ethyl Alcohol and Tobacco Cell Effects on Growth of Human Oral Keratinocytes and Human Papillomavirus 16 Entry as a Function of Furin

Background: Reported are increased risks for malignant transformation in human oral keratinocytes (HOK) from ethyl alcohol (ETOH), tobacco products or human papilloma virus oncogenic subtype 16 (HPV 16) infections. We examined various HOK cell responses to these factors to show inhibitors of epidermal growth factor receptor (EGFR) also inhibits furin;proprotein convertase (FC) and HPV 16 entry in HOK. Methods: Immortalized HOK by HPV 16 (HPV 16B) or human telomerase (hTERT);primary foreskin keratinocytes (NHFK), primary HOK, buccal keratinocytes (NHBK) and oral SCC-25 were treated with dibenz[a,l]pyrene (DBP), anthraquinone;nitrosamine (NNAL) or ethyl alcohol (ETOH) and acetaldehyde (AA). ETOH was tested for synthesis of malondialdehyde (MDA) and alcohol dehydrogenase expression (ADH). ETOH, and PAH were evaluated by Western immunoblot for oncogene changes, and phosphorylated EGFR expression. Inhibition of EGFR by WZ4002 and Erlotinib and/or carcinogens effect on HPV 16 entry were studied. A green fluorescent pseudovirus (PsV);chloromethylketone (CMK) an inhibitor of furin activity and Western immunoblot of furin cell distribution further characterized HPV 16 entry. Results: ETOH (10 μM) increased expression of phosphorylated EGFR and HPV 16 entry through furin activity, and membrane, nuclear and cytoskeletal accumulations. CMK suppressed HPV 16 entry and blockage of ADH while aldehyde dehydrogenase (ALDH) enhanced HPV 16 entry. Similarly PAH, DBP (4-8 nM), anthraquinone (98 nM) and NNAL (6.9 μM) enhanced HPV 16 entry through furin activity and membrane, nuclear and cytoskeletal accumulations. Furthermore, WZ4002 and Erlotinib suppressed expressions of phosphorylated EGFR, FC activity, and HPV 16 entry. ETOH and DBP treatments also enhanced expressions of protease activated receptor-1 (PAR-1), and p21waf1 while depressed p16 and p27KIP1 expressions in HOK/HPV 16B cells. Conclusion: EGFR inhibitors are candidates for suppression of alcohol and tobacco effects on EGFR phosphorylated expression;keratinocyte growth, and HPV 16 entry and prevention treatment for HPV related diseases.

Preparation and Evaluation of Dual Targeting Nanoparticles for Oral Cancer

A dual-receptor targeting delivery system based on acid-cleavage hydrazone bond was developed in the study. The characters of CMCS-hyd-CUR-EGFR-mAb were identified. The in vitro release studies revealed that this drug delivery system was acid-sensitive, and the self-assembled nanoparticles which were spherical. The in vitro results indicated that the dual-receptor targeting nanoparticles could be faster internalized into the Cal-27 cells via receptor-mediated endocytosis, which exhibited better antitumor activity than the one-receptor nanoparticles. The experimental results clearly reveal that CMCS-hyd-CUR-EGFR mAb provides a novel way for drug delivery in oral cancer treatment.

Arp2/3 Complex Is Expressed in Oral Squamous Cell Carcinoma: An Immunohistochemical Study of 88 Cases

Background: We investigated the immunohistochemical expression of Arp2/3 complex in 88 oral squamous cell carcinomas (OSCCs) and correlated it with the tumor type and grade. Materials and methods: This is a retrospective study of 88 cases of OSCCs (9 verrucous carcinomas, 4 carcinomas in situ, 37 well differentiated OSCCs, 32 moderately differentiated OSCCs, and 6 poorly differentiated OSCCs), stained with mouse monoclonal antibodies for Arp2 and Arp3. Staining was graded according to the extent of positivity by two observers. Results: Arp2 positivity was seen in 79 (89.77%) cases and Arp3 positivity in 78 (88.63%) cases. In positive cases, Arp2 expression was graded as (+) in 12.5%, (++) in 28.4% and (+++) in 48.86% of the cases;Arp3 was graded as (+) in 36.36%, (++) in 30.68% and (+++) in 21.59% of the cases. Conclusion: Expression of Arp2 and Arp3, indicative of the formation of the Arp2/3 complex, was seen in most OSCCs examined. Although no statistically significant correlation was found between the immunohistochemical expression of Arp2/Arp3 and tumor type or grade, the more common expression in invasive OSCCs than in non-invasive verrucous carcinomas and carcinomas in situ is indicative of its involvement in the biological behaviour of the lesions. The availability of Arp2/3 inhibitors that could be used in clinical practice warrants further study of the expression of Arp2/3 in OSCC.

Effect of westernization on oral health among college students of Udaipur City,India

Background:There is overwhelming evidence that periodontal disease and dental caries affect the majority of populations and that western culture and lifestyle may have a profound influence on oral health,especially in adults.The present study was performed to determine the effect of westernization on the oral health of college students of Udaipur City,Rajasthan.Methods:A descriptive cross-sectional study was conducted among students attending various professional and non-professional bachelor's degree colleges of Udaipur City,Rajasthan,India,from March 2013 to May 2013.Eight hundred students were selected based on a two-stage random sampling procedure.Westernization was assessed by a self-administered structured questionnaire.Periodontal status,dental caries status and malocclusion were assessed according to World Health Organization(WHO) criteria(1997).Statistical analysis was performed using Chi-square and Multivariate logistic regression.The confidence level and level of significance were set at 95% and 5%,respectively.Results:The present study suggested that adverse habits,listening to English music and preferring English food had a significant association with dental caries and periodontal diseases.Malocclusion also showed a significant relationship with consuming English food for snacks and desserts.Multivariate analysis revealed a significantly greater odds ratio(OR) for periodontal disease and dental caries among those who preferred English food for lunch.Conclusion:Based on the results of the present study,there is an association between westernization and oral health.