Healthy Oral Mucosa Research Articles

Photobiomodulation Therapy in the Prevention of Chemotherapy Induced Oral Mucositis in Children with Acute Myeloid Leukemia: A Randomized, Double-blind, Clinical trial.

Many preventive modalities have been advocated to reduce the incidence and severity of chemotherapy-induced Oral Mucositis (OM). Photobiomodulation, also known as Low-Level laser. Therapy, has shown promising results. This clinical trial was performed to evaluate the effectiveness of Photobiomodulation (PBM) for the prevention of chemotherapy-induced oral mucositis (OM) in children with Acute Myeloid Leukemia (AML). A double-blinded randomized clinical trial was performed, including forty-two children diagnosed with Acute Myeloid Leukemia (AML) admitted to receiving chemotherapy (CT) at Children's Cancer Hospital 57357, Cairo, Egypt. Patients were randomly assigned to either the control or test group. Both groups followed the standard preventive protocol for oral mucositis. In the control group I sham therapy, while in the Laser group I had Photobiomodulation sessions for five consecutive days once a day performed by Diode laser (Sirrolaser Blue ™, USA) 660nm, 15watt, 10 s. Laser sessions began on day 1 of chemotherapy and were repeated on days 2,3, 4, and 5. The response to both groups was evaluated according to the development of oral mucositis; graded using the classification criteria of the World Health Organization (WHO) and the National Cancer Institute scale (NCI - Common Terminology Criteria for Adverse Events, version 4.0). Incidence of oral mucositis was evaluated from baseline to 5, 12, 19, and 30 days after treatment. A clinical significant difference between the laser and control group was observed. The laser group showed a higher prevalence of no oral mucositis with decreased incidence of OM, while on day 12, the control group showed a higher prevalence of grades I and II (P-value <0.001, effect size = 0.796). However, the laser group showed higher prevalence of mucosa free of OM; on day 19, the control group showed higher rates of grades II and III, (P-value <0.001, effect size = 0.670). while on day 30 there was higher rates of grades I and II (P-value <0.001, effect size = 0.576) with higher rates of healthy oral mucosa in the laser group. Photobiomodulation effectively reduces the incidence and severity of chemotherapy-induced oral mucositis in pediatric patients diagnosed with AML.

Effect of short-term fasting on the cisplatin activity in human oral squamous cell carcinoma cell line HN5 and chemotherapy side effects

BackgroundKetogenic interventions like short-term fasting show potential as complementary therapies to enhance the effectiveness of chemotherapy for cancer. However, the specific effects of fasting on head and neck squamous cell carcinoma (HNSCC) cells and healthy oral mucosa cells during these treatments are not well understood. This study investigates whether short-term fasting can differentially impact HNSCC cell survival and viability compared to healthy keratinocytes while undergoing standard chemotherapy regimens.MethodsThis study investigated the effects of fasting on cell viability in HN5 cell line and healthy oral keratinocyte cells. The HN5 cell line, derived from human tongue squamous cell carcinoma, and primary human keratinocytes isolated from the basal layer of gingival epithelium were divided into three groups: (1) control, (2) treated with the standard chemotherapeutic agent cisplatin, and (3) treated with cisplatin under fasting conditions achieved through 48-hour glucose restriction mimicking the blood glucose levels of fasted individuals. Cell proliferation was assessed at 48 and 72 h using the MTT assay, a colorimetric method based on mitochondrial dehydrogenase activity. Flow cytometry analysis with specific apoptosis and necrosis markers distinguished between early and late apoptotic, necrotic, and viable cells.ResultsCell viability in HN5 and healthy keratinocyte cells decreased in cisplatin with low glucose groups compared to cisplatin and control groups. The same results were observed for healthy keratinocyte cells; only a decrease in cell viability in cisplatin groups compared to control groups was observed, which was not statistically significant. Cell apoptosis in HN5 and healthy keratinocyte cells increased in cisplatin with low glucose groups compared to cisplatin and control groups. In healthy keratinocyte cells, the cisplatin with low glucose group showed an impressive increase in necrosis, late apoptosis, and early apoptosis and a significant decrease in live cells compared with other groups.ConclusionThis study revealed that short-term fasting chemotherapy significantly improved HNSCC cell line apoptosis and necrosis.

Development of an Ex Vivo Functional Assay for Prediction of Irradiation Related Toxicity in Healthy Oral Mucosa Tissue.

Radiotherapy in the head-and-neck area is one of the main curative treatment options. However, this comes at the cost of varying levels of normal tissue toxicity, affecting up to 80% of patients. Mucositis can cause pain, weight loss and treatment delays, leading to worse outcomes and a decreased quality of life. Therefore, there is an urgent need for an approach to predicting normal mucosal responses in patients prior to treatment. We here describe an assay to detect irradiation responses in healthy oral mucosa tissue. Mucosa specimens from the oral cavity were obtained after surgical resection, cut into thin slices, irradiated and cultured for three days. Seven samples were irradiated with X-ray, and three additional samples were irradiated with both X-ray and protons. Healthy oral mucosa tissue slices maintained normal morphology and viability for three days. We measured a dose-dependent response to X-ray irradiation and compared X-ray and proton irradiation in the same mucosa sample using standardized automated image analysis. Furthermore, increased levels of inflammation-inducing factors-major drivers of mucositis development-could be detected after irradiation. This model can be utilized for investigating mechanistic aspects of mucositis development and can be developed into an assay to predict radiation-induced toxicity in normal mucosa.

The Immune Checkpoint BTLA in Oral Cancer: Expression Analysis and Its Correlation to Other Immune Modulators.

In oral squamous cell carcinoma (OSCC) tissues, an immunotolerant situation triggered by immune checkpoints (ICPs) can be observed. Immune checkpoint inhibitors (ICIs) against the PD1/PD-L axis are used with impressive success. However, the response rate is low and the development of acquired resistance to ICI treatment can be observed. Therefore, new treatment strategies especially involving immunological combination therapies need to be developed. The novel negative immune checkpoint BTLA has been suggested as a potential biomarker and target for antibody-based immunotherapy. Moreover, improved response rates could be displayed for tumor patients when antibodies directed against BTLA were used in combination with anti-PD1/PD-L1 therapies. The aim of the study was to check whether the immune checkpoint BTLA is overexpressed in OSCC tissues compared to healthy oral mucosa (NOM) and could be a potential diagnostic biomarker and immunological target in OSCC. In addition, correlation analyses with the expression of other checkpoints should clarify more precisely whether combination therapies are potentially useful for the treatment of OSCC. A total of 207 tissue samples divided into 2 groups were included in the study. The test group comprised 102 tissue samples of OSCC. Oral mucosal tissue from 105 healthy volunteers (NOM) served as the control group. The expression of two isoforms of BTLA (BTLA-1/2), as well as PD1, PD-L1/2 and CD96 was analyzed by RT-qPCR. Additionally, BTLA and CD96 proteins were detected by IHC. Expression levels were compared between the two groups, the relative differences were calculated, and statistical relevance was determined. Furthermore, the expression rates of the immune checkpoints were correlated to each other. BTLA expression was significantly increased in OSCC compared to NOM (pBTLA_1 = 0.003; pBTLA_2 = 0.0001, pIHC = 0.003). The expression of PD1, its ligands PD-L1 and PD-L2, as well as CD96, were also significantly increased in OSCC (p ≤ 0.001). There was a strong positive correlation between BTLA expression and that of the other checkpoints (p < 0.001; ρ ≥ 0.5). BTLA is overexpressed in OSCC and appears to be a relevant local immune checkpoint in OSCC. Thus, antibodies directed against BTLA could be potential candidates for immunotherapies, especially in combination with ICI against the PD1/PD-L axis and CD96.

Advancements in diagnosing oral potentially malignant disorders: leveraging Vision transformers for multi-class detection

ObjectivesDiagnosing oral potentially malignant disorders (OPMD) is critical to prevent oral cancer. This study aims to automatically detect and classify the most common pre-malignant oral lesions, such as leukoplakia and oral lichen planus (OLP), and distinguish them from oral squamous cell carcinomas (OSCC) and healthy oral mucosa on clinical photographs using vision transformers.Methods4,161 photographs of healthy mucosa, leukoplakia, OLP, and OSCC were included. Findings were annotated pixel-wise and reviewed by three clinicians. The photographs were divided into 3,337 for training and validation and 824 for testing. The training and validation images were further divided into five folds with stratification. A Mask R-CNN with a Swin Transformer was trained five times with cross-validation, and the held-out test split was used to evaluate the model performance. The precision, F1-score, sensitivity, specificity, and accuracy were calculated. The area under the receiver operating characteristics curve (AUC) and the confusion matrix of the most effective model were presented.ResultsThe detection of OSCC with the employed model yielded an F1 of 0.852 and AUC of 0.974. The detection of OLP had an F1 of 0.825 and AUC of 0.948. For leukoplakia the F1 was 0.796 and the AUC was 0.938.ConclusionsOSCC were effectively detected with the employed model, whereas the detection of OLP and leukoplakia was moderately effective.Clinical relevanceOral cancer is often detected in advanced stages. The demonstrated technology may support the detection and observation of OPMD to lower the disease burden and identify malignant oral cavity lesions earlier.

Differential expression of DNA fragmentation factor 45 in leukoplakia and oral squamous cell carcinoma and its correlation with TNF-α levels

BackgroundDNA fragmentation factor (DFF) - 45, a heterodimeric protein, involved in apoptotic processes is found to play important role in various human cancers. Its role in the development and progression of oral cavity squamous cell carcinoma (OSCC) is still unclear. Tumour necrosis factor-α (TNF-α) has been found to be higher in OSCC, yet, the mechanism of circumventing the antitumor effect of this cytokine is not understood. Thus, the aim of this study was to evaluate the expression of DFF-45 in samples of OSCC, leukoplakia, and healthy oral mucosa (control) and correlate it with salivary and serum TNF-α. Materials and methodsSurgically excised specimens from patients with primary OSCC (n = 20) leukoplakia (n = 30) and normal oral mucosa (n = 25) were immuno stained for DFF-45. The relationship of DFF-45expression with clinical parameters and histological grading was also evaluated. TNF-α levels were measured in serum and saliva collected from study subjects using cytokine ELISA technique. ResultsThe expression of DFF-45 in OSCC was either absent or mild, while a significantly up-regulated expression of this protein was observed in leukoplakia lesions. TNF-α levels in both saliva and serum were significantly higher in OSCC group compared to control and leukoplakia. We observed a negative correlation between TNF-α levels and DFF-45 expression in OSCC, similar observation was found with histological staging of OSCC. ConclusionThe absence of DFF-45 in oral cancer lesion suggests that this protein may suppress tumour progression. Down regulation of DFF-45 by tumor cells may be a mechanism to evade the anti-cancer effect of TNF-α and further investigations may unravel its role into malignant transition.

Expression of Toll-like receptors in oral squamous cell carcinoma.

Almost 380,000 new cases of oral cancer were reported worldwide in 2020. Oral squamous cell carcinoma (OSCC) accounts for 90% of all types of oral cancers. Emerging studies have shown association of Toll-like receptors (TLRs) in carcinogenesis. The present study aimed to investigate the expression levels and tissue localization of TRL1 to TRL10 and NF-κB between OSCC and healthy oral mucosa, as well as effect of Candida colonization in TRL expression in OSCC. Full thickness biopsies and microbial samples from 30 newly diagnosed primary OSCC patients and 26 health controls were collected. The expression of TLR1 to TLR10 and NF-κB was analyzed by immunohistochemistry. Microbial samples were collected from oral mucosa to detect Candida. OSCC epithelium showed lower staining intensity of TRL1, TRL2 TRL5, and TRL8 as compared to healthy controls. Similarly, staining intensity of TRL3, TRL4, TRL7, and TRL8 were significantly decreased in basement membrane (BM) zone. Likewise, OSCC endothelium showed lower staining intensity of TLR4, TLR7 and TLR8. Expression of NF-κB was significantly stronger in normal healthy tissue compared to OSCC sample. Positive correlation was found between the expression of NF-κB, TRL9 and TRL10 in basal layer of the infiltrative zone OSCC samples (P = 0.04 and P = 0.002, respectively). Significant increase in TRL4 was seen in BM zone of sample colonized with Candida (P = 0.01). According to the limited number of samples, our data indicates downregulation of TLRs and NF-κB in OSCC, and upregulation of TLR4 expression with presence of Candida.

Abstract 6036: Flash proton radiotherapy mitigates radiation-induced salivary gland dysfunction and oral mucositis in mice

Abstract Background- Recent studies have reported that ultra-high dose rate “FLASH” Proton radiation therapy (F-PRT) decreases normal tissue toxicity while maintaining tumor-controlling efficacy compared to Standard Proton RT (S-PRT), that are used for patient treatments. However, although highly efficacious in eliminating tumors, damage to the otherwise healthy salivary glands and oral mucosa is often unavoidable in patients with head and neck cancer, leaving patients with lifelong xerostomia and other comorbidities. Aim- In this study, we aimed to investigate the effect of F-PRT on radiation-induced oral mucositis and salivary gland dysfunction and in controlling orthotopic tumor growth. Methods- The head and neck area of C57Bl/6 mice was irradiated with a single dose of 16 Gy or a fractionated dose of 8 Gy x 3 of F-PRT (128Gy/s) or S-PRT (0.95 Gy/s). Oral mucositis was analyzed by histopathological examination. Radiation-induced xerostomia was studied by measuring the saliva flow rate of mice. To examine the ability of F-PRT to control orthotopic head and neck tumors, tongue tumors were generated in mice and then irradiated with F-PRT/S-PRT. Results- Following irradiation with a single dose or a fractionated dose, saliva flow was reduced by both treatments. However, the F-PRT-treated mice showed a significant improvement in salivary flow at 14 days (p&amp;lt;0.05) and 28 days (p&amp;lt;0.005) post irradiation. Expression of AQP5 was found to be significantly down-regulated at 2, 5, 10, and 14 days post irradiation with S-PRT. However, mice irradiated with F-PRT showed a significant restoration of the AQP5 expression post-RT. Oral mucositis started to appear on day 14, which showed more severity on day 28 post-irradiation with S-PRT. Histopathological analysis showed the presence of lingual gland atrophy only in the tongue of mice treated with S-PRT 28 and 60 days post-irradiation. F-PRT ameliorates radiation-induced jawbone loss compared to S-PRT as observed in mice irradiated either with a single dose or in a fractionated regime. The surviving fraction of F-PRT-treated mice with orthotopic tongue tumors was significantly ameliorated compared to S-PRT-treated mice. Conclusion- This study demonstrates that F-PRT minimizes radiation-induced normal tissue toxicity after irradiation in mice’s head and neck region. Moreover, the ability of F-PRT to control orthotopic head and neck tumors further determines the efficacy of this modality for clinical applications. Citation Format: Priyanka Chowdhury, Anastasia Velalopoulou, Ioannis Verginadis, George Morcos, Michele Kim, James Metz, Lei Dong, Alexander Lin, Constantinos Koumenis. Flash proton radiotherapy mitigates radiation-induced salivary gland dysfunction and oral mucositis in mice [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 6036.

Nlrp3 inflammasome drives regulatory T cell depletion to accelerate periapical bone erosion.

Apical periodontitis is an inflammatory disorder triggered by an immune response to bacterial infection, leading to the periapical tissue damage and alveolar resorption. However, the underlying mechanisms driving this process remain elusive, due to the complex and interconnected immune microenvironment within the local lesion site. In this study, the influence of Nlrp3 inflammasome-mediated immune response on the apical periodontitis was investigated. RNA sequencing, immunohistochemistry and ELISA assay were performed to investigate the activation of Nlrp3 inflammasome signalling pathways in the human periapical tissues, including radicular cysts, periapical granulomas and healthy oral mucosa. A mouse model of apical periodontitis was established to study the role of Nlrp3 knockout in periapical bone resorption and Treg cell stability, and the underlying mechanism was explored through invitro experiments. Invivo Treg cell adoptive transfer was performed to investigate the effects of Treg cells on the progression of apical periodontitis. Our findings find that the hyperactivated Nlrp3 inflammasome is present in human periapical lesions and plays a vital role in the immune-related periapical bone loss. Using a mouse model of apical periodontitis, we observe that Nlrp3 deficiency is resistant to bone resorption. This protection was accompanied by elevated generation and infiltration of local Treg cells that displayed a notable ability to suppress RANKL-dependent osteoclast differentiation. In terms of the mechanism of action, Nlrp3 deficiency directly inhibits the osteoclast differentiation and bone loss through JNK/MAPK and NF-κB pathways. In addition, Nlrp3 induces pyroptosis in the stem cells from apical papilla (SCAPs), and the subsequent release of cytokines affects the stability of Treg cell in periapical lesions, leading indirectly to enhanced bone resorption. In turn, adoptive transfer of both Nlrp3-deficient and wild-type Treg cells effectively prevent the bone erosion during apical periodontitis. Together, our data identify that the Nlrp3 inflammasome modulates the Treg cell stability and osteoclastogenesis in the periapical inflammatory microenvironment, thus determining the progression of bone erosion.

A Novel Prehydrated Porcine-Derived Acellular Dermal Matrix: A Histological and Clinical Evaluation.

It is well known that soft tissue quality and quantity around dental implants is of paramount importance for later peri-implant health. For this purpose, the clinical and histological outcomes of the peri-implant mucosa, following soft tissue augmentation for soft tissue improvement with a novel prehydrated porcine acellular dermal matrix graft (PPADMG) in conjunction with simultaneous implant placement, were evaluated in this case series. Twenty-two patients were included in the study. They underwent a late implant placement protocol combined with PPADMG for soft tissue augmentation. A punch biopsy was taken at the time of uncovery of the submerged healed implant after a mean of 157 days healing time. Supracrestal soft tissue height (STH) was measured at the time of implant placement and uncovery. All sites showed a clinical increase in STH. The histological structure of the biopsies resembled a similar structure as found in the healthy oral mucosa. No unexpected tissue reactions could be found. Within the limits of this clinical and histological study, it may be concluded that STH improvement with this novel porcine-derived acellular dermal matrix, in combination with simultaneous implant placement, is a viable option to create a peri-implant tissue thickness and stability.

Mast cells in oral lichen planus and oral lichenoid lesions related to dental amalgam contact.

The aim of this study was to analyze the expression of mast cell markers toluidine blue, c-kit, and tryptase and presence of mononuclear inflammatory cells in oral lichen planus (OLP) and oral lichenoid lesions related to dental amalgam. Nineteen specimens of OLP, OLLC, and healthy oral mucosa were selected. Mononuclear inflammatory cells were analyzed. Histochemical and immunohistochemical analyses were performed using toluidine blue, anti-c-kit and anti-tryptase reagents, and the results were quantified in areas A and B of connective tissue. Mast cells of all OLP and OLLC samples were positive for toluidine blue, c-kit, and tryptase. The density of toluidine blue+, c-kit+ and tryptase+ mast cells was higher in tissue with OLP and OLLC compared with healthy controls (p < 0.05). No difference was noted in mast cells density between OLP and OLLC (p > 0.05). The density of tryptase+ mast cells was higher in the subepithelial region (area A) than the region below it (Area B) in OLLC (p = 0.047). The mononuclear inflammatory cell density was higher in OLLC compared to OLP, but without statistical significance (p > 0.05). A positive statistical correlation was found between mononuclear immune cells and density of c-kit+ and tryptase+ mast cells in OLP (r = 0.943 and r = 0.886, respectively). Our data demonstrate that the etiopathogenesis process of OLP and OLLC modulates the expansion and degranulation of mast cells; mast cells density, however, was similar between OLP and OLLC. The distribution of mast cells appears to vary along the lamina propria.

Systematic Prediction of Minor Histocompatibility Antigens to Inform GvHD Outcomes after Allogeneic Stem Cell Transplantation

T cell alloreactivity against minor histocompatibility antigens (mHAgs), polymorphic peptides resulting from donor-recipient (D-R) disparity at sites of genetic polymorphisms (SNPs, indels, frameshifts), is at the core of the therapeutic effect of allogeneic hematopoietic stem cell transplantation (allo-HSCT). Despite the crucial role of mHAgs in graft-versus-leukemia and graft-versus-host (GvHD) reactions, it has not been possible thus far to consistently link patient-specific mHAg repertoires to clinical outcomes. As a result, only D-R HLA matching and the activity of GvHD prophylaxis strategies are currently available to help clinicians in this challenge. We hypothesized that post-transplant outcomes could be impacted by genome-wide mHAg load, delineated in an organ- and malignancy-specific fashion. We have therefore devised an analytic framework to systematically identify autosomal and Y-encoded mHAgs, based on the integration of polymorphism detection by whole-exome sequencing (WES) of germline DNA from D-R pairs together with organ-specific transcriptional- and proteome-level expression together with HLA class I epitope prediction. A critical component of the pipeline was building a reliable expression atlas for acute or chronic GvHD-targeted tissues. To capture less frequent but biologically relevant cell types consistently underestimated in bulk RNA expression profiles, we evaluated multiple external single-cell datasets of healthy human skin, liver, lung, GI, oral mucosa, and lacrimal gland. For each tissue, we merged the corresponding single-cell data sets and then clustered and annotated tissue-resident cell types, for which specific expressed genes were then identified, totaling 13,512 genes expressed in ≥1 GvHD target tissue. To determine how mHAg load related to GvHD risk, we applied our mHAg pipeline to paired WES (≥85% target base coverage at ≥20x depth) generated from 220 D-R pairs treated with a matched-related donor allo-HSCT for AML/MDS. We considered the impact of diagnosis, prognostic risk score, status at transplant, conditioning regimen, and graft source against median mHAg load on risk for acute and chronic GvHD. Across these patients, only total autosomal mHAg load &amp;gt; median was associated with increased risk of developing grade II-IV aGvHD (HR = 2.54, 95% confidence interval: 1.03 - 6.26, p = 0.043). While the total autosomal mHAg load was not associated with development of cGvHD, subgroup analysis of 55 FàM transplants revealed that a combined score of autosomal and Y-encoded mHAgs &amp;gt; median was associated with development of NIH moderate/severe cGvHD (p = 0.042). We next asked whether organ-specific mHAg load could predict the risk for organ-restricted acute or chronic GvHD. By evaluating organ-specific GvHD occurrence across deciles of mHAg load, we identified 2 distinct patterns of association. In the case of lung cGvHD, all 14 positive cases occurred in patients with a lung-specific autosomal mHAg load &amp;gt; median. By logistic regression modelling, lung mHAg load &amp;gt; median was the sole factor associated with increased risk of lung cGvHD (OR = 17.22, 95% CI: 3.18 - 321.63, p = 0.008). In contrast, for aGvHD of the liver, we observed an opposite pattern, with 12 of 13 affected patients having a liver mHAg load &amp;lt; median. We hypothesized that a limited set of immunodominant liver mHAgs might drive the clinical manifestations in these patients. By analyzing the mHAg landscape of patients affected by liver aGvHD, we observed an enriched representation of 7 polymorphisms in genes with preferential expression in liver. Notably, these SNPs were less frequent in patients developing aGvHD without liver involvement (p = 0.002) and co-occurred at a lower frequency in patients experiencing liver cGvHD (p = 0.0002). For one patient experiencing liver aGvHD, we could trace ex vivo T cells specific for 1 (of the 3 predicted) driver liver mHAgs. Overall, our findings indicate that molecular characterization of D-R pairs to define GvHD risk could facilitate the design of personalized post-HSCT treatments to minimize this highly morbid condition, including incorporation of additional immunosuppressive therapies, introduced early for high-risk patients, or reduced-intensity approaches in low-risk settings. This is particularly valuable for lung GvHD, which is associated with the highest morbidity and mortality.

PRF Lysates Enhance the Proliferation and Migration of Oral Squamous Carcinoma Cell Lines.

Platelet-rich fibrin (PRF) is an autologous fibrin-rich matrix where activated platelets and leucocytes accumulate. PRF has a wide spectrum of clinical indications with the overall aim of supporting tissue regeneration which in dentistry includes the healing of healthy oral mucosa with epithelial cells. In oral squamous cell carcinoma lesions, however, epithelial cells undergo malignant transformation, indicated by their unrestricted proliferation and migration potential, which should not be further enhanced by a wound-healing formula. Yet, little is known about how oral squamous cell carcinomas respond to PRF lysates. The aim of the present study was, therefore, to test the capacity of PRF lysates to change the transcriptome of HSC2 oral squamous carcinoma cells and perform bioassays to support the findings. Based on the RNAseq analysis, PRF lysates caused an increase in the genes functionally linked to cell replication and migration. In support of this screening approach, PRF lysates enhanced the proliferation of HSC2 oral squamous carcinoma cells, as indicated by 3[H]-thymidine incorporation, cell counting, and the expression of proliferation-related genes. Moreover, PRF lysates sped up cell migration in a scratch assay requiring actin polymerization. Taken together, our data showing that PRF lysates are mitogenic and stimulate motility of oral squamous carcinoma cell lines could be an indication that treatment with PRF in cases of oral carcinoma should be carefully considered.

Combined Biomarker System Predicts Prognosis in Patients with Metastatic Oral Squamous Cell Carcinoma.

Metastatic oral squamous cell carcinoma (OSCC) is associated with poor patient prognosis. Metastasis is a complex process involving various proteins, tumor cell alterations, including changes attributable to the epithelial-to-mesenchymal transition (EMT) process, and interactions with the tumor microenvironment (TME). In this study, we investigate a combined protein marker system consisting of connexin 43 (Cx43), EMMPRIN (CD147), E-cadherin, and vimentin, with a focus on their roles in the invasive metastatic progression of OSCC and their potential utility in predicting prognosis. We conducted an immunohistochemical analysis to assess the protein expression profiles of Cx43, EMMPRIN, E-cadherin, and vimentin using tissue samples obtained from 24 OSCC patients. The metastatic process was mapped through different regions of interest (ROIs), including adjacent healthy oral mucosa (OM), center of primary OSCC, invasive front (IF), and local cervical lymph node metastases (LNM). The primary clinical endpoints were disease-free survival (DFS) and overall survival (OS). Substantial changes in the expression profiles of the different marker proteins were observed among the different ROIs, with all p-values < 0.05, signifying statistical significance. Multivariable Cox regression analysis results showed a significant effect of increased EMMPRIN expression toward the IF on DFS (p = 0.019) and OS (p = 0.023). Furthermore, the combined predictive analysis showed a significant predictive value of the marker system for DFS (p = 0.0017) and OS (p = 0.00044). The combined marker system exhibited a significant ability to predict patient prognosis. An increase in EMMPRIN expression toward the IF showed the strongest effect and could be an interesting new antimetastatic therapy approach.

ХАРАКТЕРИСТИКА ЦИТОЛОГИЧЕСКОГО СОСТОЯНИЯ ПОЛОСТИ РТА У БОЛЬНЫХ ЭРОЗИВНО-ЯЗВЕННОЙ ФОРМОЙ ПЛОСКОГО ЛИШАЯ

One of the important diagnostic methods for erosive and ulcerative lesions of the oral mucosa is the cytological method. Cytological examination is a method of laboratory diagnostics, which allows to identify qualitative and quantitative changes in cells, the presence of intra- and extracellular inclusions and microorganisms. The choice of the cytological method is explained by the information content, objectivity and sufficient simplicity, and the obtained cytological data clearly correlate with the state of the oral mucosa. A cytological study was carried out in individuals with healthy oral mucosa and in patients with erosive- ulcerative form of lichen planus before and after treatment. Cytological studies have proven a violation of the cellular composition of the epithelium of the buccal mucosa in patients with erosive-ulcerative form of lichen planus, the choice of treatment plays a significant role in the timing of erosion epithelialization, recurrence and prevention of complications.

Hypovitaminosis D, objective oral dryness, and fungal hyphae as three precipitating factors for a subset of secondary burning mouth syndrome

ObjectiveElucidating the concurrence and interdependence of three precipitating factors as contributors of a subset of secondary burning mouth syndrome (BMS), which is defined having detectable precipitating factors. Design47 secondary BMS and 15 non-BMS cases were sourced from medical records of an Oral Pathology Specialty Clinic in Canada (2017–2021). Each case had Cytology, Hematology, and Sialometry tests to detail the state of three precipitating factors (the presence of fungal hyphae, hypovitaminosis D, and objective oral dryness). Three factors were compared between secondary BMS and non-BMS groups independently, in pairs, and as a triple-factor by Fisher's exact tests, Contingency Coefficients, and Logistic Regressions. ResultsRates of objective oral dryness (89.36%) and hypovitaminosis D (74.47%) in the secondary BMS group significantly differ from the non-BMS group (p = 0.0013, p = 0.0016). No difference was found in the incidence of fungal hyphae between BMS (91.49%) and non-BMS groups (p = 0.0881). Rates of three precipitating factors in pairs and as a triple-factor within the secondary BMS group significantly differ from the non-BMS group (p-values from 0.0011 to <0.0001). Their significant correlations with secondary BMS are found independently (excluding fungal hyphae), in pairs, and as a triple-factor (C-values from 0.371 to 0.461, p-values from 0.002 to <0.001). The highest C-value belongs to the triple-factor. Objective oral dryness (p = 0.009) and hypovitaminosis D (p = 0.008) are confirmed as significant predictors for secondary BMS. ConclusionsThe presence of fungal hyphae contribute to a subset of secondary BMS only when coinciding with objective oral dryness, hypovitaminosis D, or both. This interdependent relationship leads to a hypothesis that hypovitaminosis D, which is commonly called “a low value of vitamin D”, and objective oral dryness make an oral environment conducive to insidious Candida invasion, which is an intermediate status of the host-fungal interaction staying between healthy oral mucosa (non-infection) and oral candidiasis (infection).

Machine Learning in the Detection of Oral Lesions With Clinical Intraoral Images.

Artificial intelligence in oncology has gained a lot of interest in recent years. Early detection of Oral squamous cell carcinoma (OSCC) is crucial for early management to attain a better prognosis and overall survival. Machine learning (ML) has also been used in oral cancer studies to explore the discrimination between clinically normal and oral cancer. A dataset comprising 360 clinical intra-oral images of OSCC, Oral Potentially Malignant Disorders (OPMDs) and clinically healthy oral mucosa were used. Clinicians trained the machine learning model with the clinical images (n=300). Roboflow software(Roboflow Inc, USA) was used to classify and annotate images along withMulti-class annotation and object detection modelstrained by two expert oral pathologists. The test dataset (n=60) of new clinical images was again evaluated by two clinicians and Roboflow. The results were tabulated and Kappa statistics was performed using SPSS v23.0 (IBM Corp., Armonk, NY). Results: Training dataset clinical images (n=300) were used to train the clinicians and Roboflow algorithm. The test dataset (n=60) of new clinical images was again evaluated by the clinicians and Roboflow. The observed outcomes revealed that the Mean Average Precision (mAP) was 25.4%, precision 29.8% and Recall 32.9%. Based on the kappa statistical analysisthe 0.7 value shows a moderate agreement between the clinicians and the machine learning model. The test dataset showed the specificity and sensitivity of the Roboflow machine learning model to be 75% and 88.9% respectively. Conclusion: In conclusion, machine learning showed promising results in the early detection of suspected lesions using clinical intraoral images and aids general dentists and patients in the detection of suspected lesions such as OPMDs and OSCC that require biopsy and immediate treatment.

Tertiary Lymphoid Structures Are Related to Inflammatory Progression and Bone Loss in Human Apical Periodontitis

IntroductionBone loss is strongly associated with the immunologic milieu in apical periodontitis (AP). Tertiary lymphoid structures (TLSs) are organized lymphoid cell aggregates that form in nonlymphoid tissues under persistent inflammatory circumstances. To date, there has been no relevant report of TLSs in periapical lesions. This work aimed to investigate the formation and potential function of TLSs in AP. MethodsTissues from human apical lesions (n = 61) and healthy oral mucosa (n = 5) were collected. Immunohistochemistry and multiplex immunofluorescence were used to detect the formation of TLSs. Correlation analyses were performed between clinical variables and TLSs. In addition, immunohistochemistry was used to evaluate the expression of interleukin-1 beta, interleukin-6, receptor activator of nuclear factor kappa-B ligand, and macrophage subsets in the apical lesions. ResultsPeriapical granulomas (n = 24) and cysts (n = 37) were identified by histologic evaluation. TLSs, composed of B-cell and T-cell clusters, developed in periapical granulomas and radicular cysts. The CXC-chemokine ligand 13, its receptor CXC-chemokine receptor 5, follicular dendritic cells, and high endothelial venules were localized in TLSs. The quantity and size of TLSs were positively associated with bone loss in AP. Moreover, proinflammatory cytokines and macrophage subsets were also substantially elevated in TLS regions of apical lesions. ConclusionsThe formation of TLSs in periapical granulomas and cysts was closely associated with persistent immune responses and bone loss in apical lesions. TLSs provide an updated insight into the complicated immune response process in AP.

Epstein-Barr virus predominance and immunological abnormalities in oral squamous cell carcinoma

BACKGROUND: Both benign and harmful head and neck disorders have been associated with the Epstein-Barr virus (EBV). Many studies have connected EBV to oral squamous cell carcinoma (OSCC). Oral squamous cell carcinoma is the most prevalent type of cancer. Fresh tissue samples from patients with OSCC were tested for the presence of Epstein-Barr virus. OBJECTIVE: To determine the frequency of EBV and IL10 expression in OSCC patients. MATERIALS AND METHODS: Fifty individuals with OSCC and 25 with clinically healthy oral mucosa were studied. In situ hybridization was used for the detection of EBV. Serum IL-10 levels were also evaluated in patients and controls using a human IL-10 ELISA Kit. RESULTS: EBV was detected in 4 healthy patients, 6 with moderately differentiated OSCC, 10 with poorly differentiated OSCC, and 19 with undifferentiated OSCC. These differences were statistically significant (p&lt;0.05). IL-10 expression was more common in OSCC diagnostic groups than healthy controls, and the difference in blood IL-10 levels between patients and controls was statistically significant (p&lt;0.01). CONCLUSION: The prevalence of EBV in OSCC suggests its possible role in oral cavity malignancy. On the other hand, IL-10 is expressed at higher levels in OSCC biopsies; such elevated concentrations may promote viral spread.

IL-10 A/G rs1800896 and TNF-α G308A rs1800629 Polymorphisms and Their Relationship with the Risk of Implant Loss in Adult Patients from Argentina

Introduction: the results of treatment in implantology have been evaluated mainly as implant survival rates in small groups of patients selected from specialized clinical or university settings. There is evidence to support polymorphisms that could be linked to some biological complications in implantology. The results may vary according to the origin or ethnic mixtures of the population studied. The objective of this study was to analyze the relationship between the polymorphisms IL-10 A/G rs1800896 and TNF-α G308A rs1800629308 and the loss of dental implants and periodontal disease. Material and method: 140 patients were selected, 10 with implant losses within a maximum period of 6 months of their placement. Each patient voluntarily consented to participate in the study (approved by CIEIS Adult Hospital Córdoba) Filiatory and clinical data were collected in a clinical history. Samples of clinically healthy oral mucosa were obtained and genotyped by PCR and RFPL. Results: The population consisted of male and female individuals in similar proportions (p=0.6121). The average age was 53.06±16.22 years, and an age variation range of 20 to 80 years. 7.7% of the patients included in the study had loss of their implant. It was observed that 62.5% of the patients who presented loss of implants were women. Of the patients with missing implants, 75% were smokers and did not consume alcohol. On the other hand, in relation to periodontal disease only 31% presented disease. In relation to genotype, patients with lost implants had 50% of the mutated allele of the SNP TNFα rs1800629, while 50% of patients with periodontal disease were carriers of the mutated allele for SNP IL-10 rs1800896. On the other hand, considering the total population under study, 31.06% of the patients presented the genotypes with the genetic variation, AG+GG, of the IL10 rs1800896 gene; while 64.07% presented the AA and GA genotypes, (both with the mutated allele) of the TNF-α G308A rs1800629 gene. No significant association (p=0.3298) was observed between IL10 rs1800896 genotypes and periodontal disease; contrary to whether there was a significant relationship of this SNP with periodontal disease (p=0.0164). Conclusion: The polymorphisms evaluated were not predictive of the failure of dental implants. However, a significant association between periodontal disease and TNF-α rs1800629 genotype could be observed. It is noteworthy that this is the first study that describes the frequency of the SNPs studied in a population of Córdoba–Argentina.