Effects of Saliva From Periodontally Healthy and Diseased Subjects on Barrier Function and the Inflammatory Response in in vitro Models of the Oral Epithelium

Background: Periodontitis is a multifactorial, bacteria-mediated chronic inflammatory disease that results in the progressive destruction of the tooth-supporting tissues. It is well-known that saliva from subjects suffering from this disease generally contains higher levels of pro-inflammatory mediators, matrix metalloproteinases (MMP), and bacteria-derived toxic products. The aim of this study was to investigate and compare the effects of saliva from periodontally healthy and diseased subjects on the barrier function and inflammatory response in in vitro models of the oral epithelium.Methods: Unstimulated saliva samples from two groups of subjects, one with a healthy periodontium (n = 12) and one with severe generalized periodontitis (n = 11), were filter-sterilized. All the saliva samples were analyzed using an immunological multiplex assay to determine the levels of various cytokines and MMPs relevant to periodontitis. The impact of saliva on epithelial barrier integrity was assessed by monitoring transepithelial electrical resistance (TER) in an oral epithelium model using the B11 keratinocyte cell line. GMSM-K oral epithelial cells were treated with saliva from both groups to determine their ability to induce the secretion of interleukin-6 (IL-6) and interleukin-8 (IL-8), as determined by an enzyme-linked immunosorbent assay (ELISA).Results: Saliva from the periodontitis subjects contained significantly higher concentrations of matrix metalloproteinase-8 (MMP-8), matrix metalloproteinase-9 (MMP-9), IL-8, and C-X-C motif chemokine ligand 1 (CXCL1) compared to saliva from the healthy subjects. Saliva from the healthy and periodontitis subjects affected cytokine secretion and TER in a similar manner. More specifically, saliva from both groups increased TER and induced IL-6 and IL-8 secretion in the in vitro oral epithelium models used.Conclusion: Independently of the presence or absence of periodontitis, saliva can increase the relative TER and the secretion of IL-6 and IL-8 in in vitro models of the oral epithelium.

Can tooth bleaching agents cause genotoxicity in the oral epithelium? A systematic review with meta-analysis

Objective: To assess, with a systematic review (SR) with meta-analysis, the occurrence of genotoxic effects on the oral epithelium after exposure to tooth bleaching agents. Material and methods: This review was performed according to the PRISMA protocols. To identify relevant studies, a systematic search was performed in the following electronic databases: PubMed, Scopus, Embase, LILACS, and Google Scholar. The research question was "Can tooth bleaching agents cause genotoxicity in the oral epithelium?”. The treatment effects were defined as the standardized mean difference (SMD) and 95% confidence intervals (CI) were established. Results: 154 studies were selected and, after screening titles and abstracts, seven full-text manuscripts were assessed for eligibility, of which four studies were included in the meta-analysis. There were no statistically significant differences in the frequency of micronuclei before and after exposure (SMD= -0.14, 95% CI, 0.98 to 0.60, p=0.74), with a Tau2 index = 1.00; Chi2 = 70.20; p-value <0.00001; and I² of 93%, indicating high heterogeneity among the studies. Conclusion: Considering the limitations of the present SR, tooth bleaching agents do not lead to genotoxic damage in the oral epithelium but with a small effect and low level of evidence. In this way, the use of tooth bleaching agents is safe on the oral mucosa but randomized clinical trials that are more standardized in all stages are required to produce more robust evidence.

O Impacto dos Sintomas Orais Gerados por Quimioterapia e Radioterapia / The Impact of Oral Symptoms Generated by Chemotherapy and Radiotherapy

Os primeiros efeitos da radioterapia e da quimioterapia antineoplásica acontecem sobre as células do epitélio oral, as quais sofrem rápida proliferação. O tamanho destes efeitos depende de muitos fatores ligados ao tratamento, ao paciente e ao tumor. No que se trata do paciente, interferem nesse processo o seu estado geral de saúde, presença de comorbidades, sexo, estado nutricional, idade, fatores sociais e psicológicos, além de hábitos deletérios e patologias orofaciais preexistentes. Esses adoecimentos na integridade bucal devem-se a veracidade de que a radioterapia e quimioterapia não são capazes de destruir as células tumorais sem lesionar células normais. O tratamento oncológico pode provocar reações adversas na cavidade oral, e é comum, em pacientes oncológicos submetidos ao tratamento antineoplásico, o desenvolvimento de agravamentos orais agudos ou tardios.---The first effects of radiotherapy and antineoplastic chemotherapy happen on the cells of the oral epithelium, which quickly notice proliferation. The size of these effects depends on many factors related to the treatment, the patient, and the tumor. With regard to the patient, this process interferes with their general health status, presence of comorbidities, gender, nutritional status, age, social and psychological factors, in addition to deleterious habits and pre-existing orofacial pathologies. These illnesses in the oral integrity are due to the veracity that radiotherapy and chemotherapy are not capable of destroying tumor cells without normal cells. Oncological treatment can cause adverse reactions in the oral cavity, and it is common, in cancer patients, to antineoplastic treatment, to develop acute or late oral aggravations.

The Effect Of Ethanol Extract From Lingzhi Fungi (Ganoderma Lucidum) Cianjur Isolate On Syndecan-1 Expressions In Kb CCL17 Oral Cancer Cell

Intercellular adhesion plays a role in cancer formation and protein has a key potential in maintaining cell adhesion, including syndecan-1. Meanwhile, oral cancer originates from the oral epithelium, which has an invasive and metastatic level. Its treatments involving chemotherapy and radiotherapy commonly leave unfavorable side effects, hence, suitable alternatives are needed. Natural ingredients are widely used as an alternative treatment for cancer, for example, Ganoderma lucidum (G. lucidum) which has anti-cancer and anti-angiogenic properties, induces apoptosis, stimulates an immune response, inhibits the degradation of Extracellular matrix (ECM), reduces inflammation, affects cell cycles, cytotoxic, and acts as an antioxidant.This study aims to determine the effect of ethanol extract from Ganoderma lucidum Cianjur isolate on syndecan-1 expression in KB CCL-17 oral cell cancer. This was an experimental study with a post-test only control group design, the treatment group used G. lucidum ethanol extract with a concentration of 2.12 μg/ml (P1), 4.24 μg/ml (P2), and 8.49 μg/ml (P3), while the positive control group used cisplatin with a concentration of 11.5 μg/ml (K1). In contrast, the negative control used aquadest (K0), while syndecan-1 expression was observed using the immunohistochemical examination.The highest syndecan-1 expansion rate was found in the treatment group with a concentration of 8.49 μg/ml. A significant difference was indicated by one-way ANOVA (p<0.05) between K0 - K1, K0 - P1, K0 - P2, K0 - P3, K1 - P1, K1 - P2, K1 - P3, P1 - P2, as well as P1 and P3. The administration of ethanol extract from G. lucidum Cianjur isolate increases syndecan-1 expression in KB CCL-17 oral cell cancer.

Possible Interference in Protein – Protein interaction as a new approach in microinhibition of respiratory pathogens on nasal– oral epithelium: An early on-screen study with reference toSARS-Cov-2–ACE2 binding interference

Upper and lower respiratory pathogens – both microbes and viruses –are responsible for very high morbidity, man-hour loss, residual long term clinical conditions and even mortality. In india only, high incidence of annual respiratory infections – both UT and LT – demands prophylactic intervention in addition to all therapeutic interventions available.The issue of respiratory infections is more pronounced now in the backdrop of nearly uncontrolled high incidences of SARS-Cov-2 affection resulting in death and damage of human lives to the extent of hundreds of millions spreading over entire world, with incidence variations from country to country. After the initial unanswered phase of spread of SARS-Cov-2 virus with attendant unseen mortalities, quickest invention of a series of unusual vaccines have stemmed the lethal progress to a very significant extent, although vaccinating each and every human subject – nearly 8 to 9 bn in supremely divided world –economically-- is an unthinkable proposition where economic disparity dictates vaccine availability and implementation.Moreover, being of highly unstable nucleic acid composition, the original virus, by now has a thick set of variants around the globe with variable clinical outcome. Given this complex background of scanty availability and inefficient implementation, there always is a need of a preventive approach which can possibly micro-fix the pathogens, including SARS-2 on nasal epithelium so as to interfere with viral [or any pathogen] entry through specified receptor gate[s] or any other ways. The present formulation is under study -- as a candidate of interference on nasal / oral mucosa for all respiratory pathogens. This brief report describes dry on-screen studies of protein – protein interaction as well as its possible interference by an amino acid Lysine. Phospholipid bilayerresponses in presence of added loads of the same essential amino acid –Lysine – showed unusual and unexplained behavior both in structural integrity as well in spatial orientation.

Detection of Epstein Barr Virus (EBV) Antigen in Oral Squamous Cell Carcinoma (OSCC) by Immunohistochemistry in Patients from KPK Province of Pakistan.

Objective: To determine the frequency of EBVLMP-1 antigen positivity in OSCC and normal oral squamous epithelial tissues (control) by immunohistochemistry (IHC) and to see the effect of age and gender in both the OSCC and control tissues. Study Design: Descriptive, Cross Sectional, Multicenter study. Setting: Pathology/ Peshawar Medical College (PMC), Peshawar Dental College and Sardar Begum Dental College, Peshawar. Period: April 2018 to September 2018. Material & Methods: Conducted on total 60 samples divided into two groups. Group A: 30 formalin fixed paraffin embedded tissue blocks of OSCC cases and Group B: 30 non neoplastic oral epithelial tissues (control). Both groups A and B slides were stained through immunohistochemistry for EBV LMP-1 monoclonal antigen. Results: 30 cases of OSCC Group A and 30 cases of non-neoplastic oral mucosa Group B were selected for the LMP-1 staining by immunohistochemistry. The mean age of OSCC was 56.5 years (±14.47 Standard deviation) with a range of 20-80 years. Male: Female ratio was 1.3:1 .OSCC commonly involved buccal mucosa. 80% (n=24/30) OSCC cases were grade 1, all the cases of OSCC (100%) showed epithelial positivity for EBV LMP-1 antigen, whole (29/30) cases of non-neoplastic oral mucosa cases showed positivity and there is no statistically significant difference in both Groups A and B. Conclusion: With the findings of EBV positivity in both the OSCC and all of the control cases; it is concluded that EBV does not play an important role in etiology of OSCC. This suggests that the infection by this ubiquitous virus (EBV) occurs at some point in the life of a person leaving LMP, protein latent in the cells of oral epithelium.

High-Risk Human Papillomavirus and Epstein–Barr Virus Coinfection: A Potential Role in Head and Neck Carcinogenesis

High-risk human papillomaviruses (HR-HPVs) and Epstein–Barr virus (EBV) are recognized oncogenic viruses involved in the development of a subset of head and neck cancers (HNCs). HR-HPVs are etiologically associated with a subset of oropharyngeal carcinomas (OPCs), whereas EBV is a recognized etiological agent of undifferentiated nasopharyngeal carcinomas (NPCs). In this review, we address epidemiological and mechanistic evidence regarding a potential cooperation between HR-HPV and EBV for HNC development. Considering that: (1) both HR-HPV and EBV infections require cofactors for carcinogenesis; and (2) both oropharyngeal and oral epithelium can be directly exposed to carcinogens, such as alcohol or tobacco smoke, we hypothesize possible interaction mechanisms. The epidemiological and experimental evidence suggests that HR-HPV/EBV cooperation for developing a subset of HNCs is plausible and warrants further investigation.

Correlation of Soluble CD44 Expression in Saliva and CD44 Protein in Oral Leukoplakia Tissues

The aim of this study was to determine whether and how pan-CD44 protein expression in leukoplakia tissues correlates with positive SolCD44 test presence and their role in oral leukoplakia. SolCD44 and total protein expression in saliva were determined using an OncAlert® Oral Cancer Rapid test. Comparison of paired associations of total protein, SolCD44, mean number of CD44 expressed epithelial layers in leukoplakia tissue, and macrophages below the basement membrane between control group and patients with leukoplakia showed statistically significant results (p < 0.0001). It is shown that the total protein indicates low or elevated risk of possible malignant transformation processes in leukoplakia. Statistically significant differences between higher total protein level and clinical forms of oral leukoplakia (p < 0.0001), as well as CD44-labeled epithelial cell layer decrease (p < 0.0001), were found. This possibly points to the onset of the stemness loss in leukoplakia tissue. CD9 antigen expression in the exosomes of the oral epithelium explained the intercellular flow of SolCD44 and other fluids in the leukoplakia area. We conclude that the OncAlert® Oral Cancer Rapid test is a valuable screening method in daily clinical practice, in terms of complementing clinical diagnostics methods and to assess the potential for early malignancy.

Spatiotemporal Expression and Functional Analysis of miRNA-22 in the Developing Secondary Palate

Objective Normal development of the embryonic orofacial region requires precise spatiotemporal coordination between numerous genes. MicroRNAs represent small, single-stranded, non-coding molecules that regulate gene expression. This study examines the role of microRNA-22 (miR-22) in murine orofacial ontogeny. Methods Spatiotemporal and differential expression of miR-22 (mmu-miR-22-3p) within the developing secondary palate was determined by in situ hybridization and quantitative real-time PCR, respectively. Bioinformatic approaches were used to predict potential mRNA targets of miR-22 and analyze their association with cellular functions indispensable for normal orofacial ontogeny. An in vitro palate organ culture system was used to assess the role of miR-22 in secondary palate development. Results There was a progressive increase in miR-22 expression from GD12.5 to GD14.5 in palatal processes. On GD12.5 and GD13.5, miR-22 was expressed in the future oral, nasal, and medial edge epithelia. On GD14.5, miR-22 expression was observed in the residual midline epithelial seam (MES), the nasal epithelium and the mesenchyme, but not in the oral epithelium. Inhibition of miR-22 activity in palate organ cultures resulted in failure of MES removal. Bioinformatic analyses revealed potential mRNA targets of miR-22 that may play significant roles in regulating apoptosis, migration, and/or convergence/extrusion, developmental processes that modulate MES removal during palatogenesis. Conclusions Results from the current study suggest a key role for miR-22 in the removal of the MES during palatogenesis and that miR-22 may represent a potential contributor to the etiology of cleft palate.

The Balance between Differentiation and Terminal Differentiation Maintains Oral Epithelial Homeostasis

The oral epithelium is one of the fastest repairing and continuously renewing tissues. Stem cell activation within the basal layer of the oral epithelium fuels the rapid proliferation of multipotent progenitors. Stem cells first undergo asymmetric cell division that requires tightly controlled and orchestrated differentiation networks to maintain the pool of stem cells while producing progenitors fated for differentiation. Rapidly expanding progenitors subsequently commit to advanced differentiation programs towards terminal differentiation, a process that regulates the structural integrity and homeostasis of the oral epithelium. Therefore, the balance between differentiation and terminal differentiation of stem cells and their progeny ensures progenitors commitment to terminal differentiation and prevents epithelial transformation and oral squamous cell carcinoma (OSCC). A recent comprehensive molecular characterization of OSCC revealed that a disruption of terminal differentiation factors is indeed a common OSCC event and is superior to oncogenic activation. Here, we discuss the role of differentiation and terminal differentiation in maintaining oral epithelial homeostasis and define terminal differentiation as a critical tumour suppressive mechanism. We further highlight factors with crucial terminal differentiation functions and detail the underlying consequences of their loss. Switching on terminal differentiation in differentiated progenitors is likely to represent an extremely promising novel avenue that may improve therapeutic interventions against OSCC.