Role of reflectance confocal microscopy for in vivo investigation of oral disorders: White, red and pigmented lesions.

In vivo reflectance confocal microscopy (RCM) is poorly investigated in oral pathology due to the peculiar anatomical and topographical oral mucosa features. A dedicated handheld confocal microscope with an intra-oral probe was developed for oral mucosa imaging. The main objective was to describe the healthy oral mucosa and the cytoarchitectural findings detectable in different oral disorders by means of the newly designed handheld confocal microscope. Secondary aim was to identify the main RCM criteria that differentiate oral lesions in order to provide algorithm for a rapid non-invasive evaluation. This observational retrospective study included all consecutive patients with oral disorders and volunteers with healthy oral mucosa who underwent RCM examination in our outpatient clinic from September 2018 to December 2021. Three different investigators examined together the RCM images to detect the key features and secondary criteria for each type of oral lesion collected. The study population included 110 patients affected by oral lesions and seven volunteers with healthy oral mucosae. A total of 15 oral disorders were imaged and divided in three main groups: white, red and pigmented lesions. Key features and secondary criteria were identified for every single type of oral disease. RCM permits a cytoarchitectural evaluation of the oral mucosae affected by inflammatory, dysplastic and neoplastic diseases, thus orienting the clinicians towards non-invasive diagnosis and enhancing the diagnostic management. The "tree diagrams" proposed allow a schematic and simplified view of confocal features for each type of oral disease, thus drastically reducing the diagnostic timing.

Identification of Candida species in the clinical laboratory: a review of conventional, commercial, and molecular techniques.

In healthy individuals, Candida species are considered commensal yeasts of the oral cavity. However, these microorganisms can also act as opportunist pathogens, particularly the so-called non-albicans Candida species that are increasingly recognized as important agents of human infection. Several surveys have documented increased rates of C. glabrata, C. tropicalis, C. guilliermondii, C. dubliniensis, C. parapsilosis, and C. krusei in local and systemic fungal infections. Some of these species are resistant to antifungal agents. Consequently, rapid and correct identification of species can play an important role in the management of candidiasis. Conventional methods for identification of Candida species are based on morphological and physiological attributes. However, accurate identification of all isolates from clinical samples is often complex and time-consuming. Hence, several manual and automated rapid commercial systems for identifying these organisms have been developed, some of which may have significant sensitivity issues. To overcome these limitations, newer molecular typing techniques have been developed that allow accurate and rapid identification of Candida species. This study reviewed the current state of identification methods for yeasts, particularly Candida species.

Variations of oral anatomy and common oral lesions.

Several topics related to the oral cavity are briefly addressed in this article, from anatomical variations that, when recognized, avoid unnecessary investigations, to diseases that affect exclusively the mouth, mucocutaneous diseases, as well as oral manifestations of systemic diseases. A complete clinical examination comprises the examination of the mouth, and this approach facilitates clinical practice, shortening the path to diagnosis in the outpatient clinic as well as with in-hospital patients. The objective of this article is to encourage the examination of the oral cavity as a useful tool in medical practice, helping to recognize diseases in this location.

Macroscopy of specimens from the head and neck.

Macroscopic examination of surgical resections from the head and neck may be difficult due to the complex anatomy of this area. Recognition of normal anatomical structures is essential for accurate assessment of the extent of a disease process. Communication with the surgical team, correct specimen orientation and sampling are critical for assessment and the importance of radiological and clinical correlation is emphasised. Tumour involvement at each subsite is highlighted with reference to where there are implications on pathological staging and the potential need for adjuvant therapy.

Comparative analysis of antigen-presenting cells in gingival tissues in healthy and periodontitis patients.

AIMS: Microbial flora of dental plaque trigger innate and adaptive immune responses. The function of antigen-presenting cells (APCs) is to bridge the innate and adaptive immune systems. The human immune system contains three main types of APCs: dendritic cells (DC) (Langerhans cells (LCs) and interstitial DCs, IDCs), macrophages and B lymphocytes. In this study, the distribution and density of all APCs in healthy and inflamed human gingival tissue were comparatively analysed. METHODS: Research was conducted on gingival biopsy specimens obtained from 55 patients and classified in three groups: healthy gingiva (control group, n=10), moderate periodontal disease (PD) (n=21) and severe PD (n=24). For APCs' identification antibodies raised against CD1a (for LCs), S100 protein (for iDCs), CD68 (for macrophages) and CD20 (for B lymphocytes) were used. RESULTS: Increased density of IDCs, macrophages and B lymphocytes in lamina propria and reduced density of LCs in the gingival epithelium were found in patients with periodontitis. Simultaneously, it was noticed an increased concentration of macrophages and B cells in the gingival epithelium in patients with PD. No statistically significant difference in the distribution and density of APC was found among patients with moderate and advanced periodontitis. CONCLUSIONS: It was hypothesised that in the periodontitis the role of antigen presentation was largely taken from LCs by the DCs, macrophages and B cells. These APCs are thought to have less protective and tolerogenic potential than LCs and this is a significant reason for alveolar bone destruction in periodontitis.

Tumour-infiltrating lymphocyte subsets and their individual prognostic impact in oral squamous cell carcinoma.

AIMS: Current understanding of oral squamous cell carcinoma (OSCC) is incomplete with regard to prognostic factors that lead to the considerable heterogeneity in treatment response and patient outcomes. We aimed to evaluate the impact of individual tumour-infiltrating lymphocyte (TIL) subsets on prognosis as a possible rationale for this, in a retrospective observational study. METHODS: Immunohistochemistry was performed to quantitatively assess cell densities of CD3+, CD20+, CD4+, CD8+ and FOXP3+TIL subsets in 50 surgically treated OSCC cases. Results were correlated with disease-free survival (DFS) and overall survival (OS). Receiver operating characteristic curve analysis and Youden index were applied to determine prognostically significant cut-off values. RESULTS: Mean counts for CD3+, CD4+, CD8+, CD20+ and FOXP3+TILs were 243, 52, 132, 53 and 116 cells per high power field, respectively. High CD8+ and low FOXP3+TIL counts, and high ratio of CD8:FOXP3 were significantly associated with longer DFS and OS, as well as with improved tumour-host interface parameters. CONCLUSIONS: Host immune response and its interaction with cancer cells have a significant impact on OSCC outcomes, with some TIL subsets being more clinically relevant than others. High cytotoxic T-cell (CD8) and low Treg (FOXP3) counts, and high cytotoxic T-cell to Treg (CD8:FOXP3) ratio are significantly associated with favourable prognosis. These results may serve as a leading point in identifying novel therapeutic agents that can redesign the tumour immune microenvironment by reducing infiltrating FOXP3-lymphocytes, and modifying their signalling pathways.

Verification and validation of the anti-PD-L1 antibody, Clone 22C3 on a laboratory-developed test.

AIMS: The first aim of this study is to compare and validate the performance of the programmed death receptor ligand 1 (PD-L1) IHC 22C3 pharmDx assay kit processed via Dako Omnis platform with the Dako Autostainer Link 48. The second aim is to examine the concordance of scoring by pathologists using the same immunohistochemistry (IHC) assay on the Dako Omnis platform and the Dako Autostainer Link 48. METHODS: Fourty-seven formalin-fixed, paraffin-embedded tissue blocks of head and neck squamous cell carcinoma tumour were stained with the PD-L1 IHC 22C3 pharmDx assay kit processed via the Dako Autostainer Link 48 and the Dako Omnis platform. Combined positive score (CPS) was ascribed by two scoring pathologists, with discordant cases provided with an agreed score. RESULTS: First, identical staining patterns were identified. Second, high agreement of PD-L1 scores when a CPS cut-off of 1 was implemented illustrated an overall agreement of 94%, positive agreement of 100% and negative agreement of 88%. Finally, results highlight an intraexaminer concordance of 89% and interexaminer concordance of 85% and 92%. CONCLUSIONS: In conclusion, we propose to open for discussion the deconstruction of the current practice of a compulsory companion diagnostic test (CDT) for a particular PD-L1 immunohistochemical assay. The implementation of laboratory developed tests as an alternative to the CDT poses as a novel and readily available method to surmount limitations posed to pathology laboratories.

Science mapping analysis of maxillofacial reconstruction over the last four decades.

PURPOSE: This paper aims to provide a bibliometric analysis of the maxillofacial reconstruction literature over 40 years and to compare the data with previous studies. METHODS: A bibliographical search for oral and maxillofacial surgery literature in maxillofacial reconstruction was conducted on Wef of Science. A graphic representation of authorship and keywords was created with VOSviewer. Mendeley and Microsoft Excel were used for tabulation and data visualization. Some statistical tests were performed with a 95 % confidence interval, which was considered significant. RESULTS: A total of 7417 articles from specialized journals were included in the study. These articles received 138,493 citations from 63,390 other studies, with an average citation count of 18.67, and a very high H-index. A total of 2375 specific keywords were analyzed, covering a wide range of topics, with two noteworthy MeSH keywords that have recently gained prominence. A total of 33 journals were included in the study, with a mean Impact Factor of 2.404, indicating a relatively high influence in the subject area. CONCLUSION: The high h-index reflects abundant and high-quality literature on maxillofacial reconstruction, with the United States leading in publication quantity. Emerging topics in maxillofacial reconstruction were discussed. These areas shape the discipline, driving advancements and offering opportunities for researchers and clinicians to contribute to progress and improve patient outcomes.

Oral and maxillofacial literature from Middle East: a bibliometric analysis and list of top-100 most cited articles.

OBJECTIVES: The aim of this work is to perform a bibliometric analysis on Middle Eastern oral and maxillofacial surgery through years and to compare the data with previous studies. A list of the top 100 most cited articles was generated. METHODS: A Pubmed bibliographic search for literature journals specialized in oral and maxillofacial surgery in the Middle East was performed. A graphic representation of authorship and keywords was created with VOSviewer. Mendeley and Microsoft Excel were used for tabulation and data visualization. A list of the top 100 most cited articles was created using Web of Science. Some statistical tests were performed with a 95% confidence interval, which was considered significant. RESULTS: A total of 6,536 articles were retrieved in fifteen selected oral and maxillofacial journals. A moderate correlation between number of publications and total population (R = 0.6052), low correlation with area (R = 0.302291), and a negligible correlation with the Human Development Index (HDI) were found (R = 0.1747). A disparity can be seen in the number of publications by country, leaded by Turkey (46.30%), Israel and Iran (13.68% each). The more common studied topic was oral surgery (25.77%), maxillofacial trauma (16.13%) and oral pathology (10.25%). CONCLUSIONS: A useful list of the top 100 most cited articles on oral and maxillofacial surgery from Middle East has been created. Middle East publications on oral and maxillofacial surgery can be considered high quality (IF = 1.879). Some issues on performing an adequate selecting of MeSH keywords were discussed.

Analytical Study Of Salivary Mmp-12 Expression In Oral Submucous Fibrosis.

BACKGROUND: Oral submucous fibrosis (OSF) is majorly a pathology pertaining to Asian population, reported most in Pakistan, India, Nepal, Bangladesh, and Taiwan. Equilibrium existing between Matrix metalloproteinases (MMP) & tissue inhibitors of matrix metalloproteinases is imperative for the normal integrity of connective tissues. However, this mechanism is disturbed in the presence of OSF, resulting in an increase in the extracellular matrix. METHODS: It is an analytical study including two groups with a total of 60 participants. The first group consists of 30 healthy participants and the other group consists of 30 patients presenting with oral submucous fibrosis. Collected samples of saliva were stored at -80 °C after centrifugation. For ELISA investigation, the procedure was performed as per manufacturer's instruction. Salivary matrix metalloproteinases- 12 concentration was estimated with the help of a standard curve. Data was analysed using SPSS 23. Mann Whitney test was applied to determine the difference existing in Matrix metalloproteinases- 12 levels between healthy and oral submucous fibrosis participants. p-value <0.05 was contemplated as significant. RESULTS: Statistical investigation indicated significant difference in Matrix metalloproteinases- 12 levels between Oral submucous fibrosis and healthy group (p<0.05). Saliva samples obtained from oral submucous fibrosis patients demonstrated raised concentrations of Matrix metalloproteinases- 12 as compared to healthy participants. CONCLUSIONS: Our study demonstrates significant upsurge in Matrix metalloproteinases- 12 expression in samples of saliva obtained from oral submucous fibrosis patients as compared to healthy individuals. Therefore, salivary Matrix metalloproteinases- 12 could serve as a useful diagnostic marker for OSF.

Porphyromonas gingivalis, the most influential pathogen in red-complex bacteria: A cross-sectional study on the relationship between bacterial count and clinical periodontal status in Japan.

BACKGROUND: Porphyromonas gingivalis is a key pathogen in microbiota associated with periodontitis. The purpose of the present study was to assess the association between salivary counts of red-complex bacteria and clinical periodontal status in a Japanese population. METHODS: A total of 977 subjects who visited a general dental clinic in Japan from 2003 to 2006 were enrolled in the study. Stimulated saliva was obtained, and the amounts of major periodontal bacteria were measured using real-time polymerase chain reaction. Probing pocket depth (PPD), bleeding on probing (BOP), and each subject's average proximal bone crest level (BCL) on dental radiographs were measured. RESULTS: The number of P. gingivalis strongly associated with percentage of 4 mm or more PPD sites, BOP positive percentage, and 1.5 mm or more BCL sites. The detection of P. gingivalis with Treponema denticola and/or Tannerella forsythia showed a high rate of three positive clinical parameters, whereas the only P. gingivalis detected group and those without P. gingivalis had a low rate of three positive clinical parameters. CONCLUSION: Among red-complex bacteria, the amount of P. gingivalis showed the strongest association with the severity of periodontal condition, and co-occurrence of P. gingivalis with T. denticola and/or T. forsythia showed heightened progression of periodontitis.

The virulence factor GroEL directs the osteogenic and adipogenic differentiation of human periodontal ligament stem cells through the involvement of JNK/MAPK and NF-κB signaling.

BACKGROUND: GroEL, a bacterial metabolite, is an important stimulator of inflammation. The aim of this study is to confirm the effect of the virulence factor GroEL on differentiation potential of periodontal ligament (PDL) stem cells (PDLSCs) and the potential mechanisms. METHODS: PDLSCs were obtained from extracted human premolars. GroEL was administered to osteogenic- and adipogenic-induced hPDLSCs. Alkaline phosphatase (ALP) staining, Alizarin Red staining and Oil Red staining were performed. Gene and protein expression were separately measured by qPCR and Western blotting. The expression and localization of activated signaling factors were confirmed by immunofluorescence staining. The inhibitors of myeloid differentiation factor 88 (MyD88, an adaptor protein of TLRs), JNK/MAPK and NF-κB signaling were used to verify their specific effects. RESULTS: First, we found that GroEL inhibited the osteogenic differentiation and enhanced the adipogenic differentiation of hPDLSCs. Next, we found that GroEL increased the expression of TLR2 and TLR4 and GroEL activated JNK/MAPK and NF-κB signaling, which can be blocked by inhibition of MyD88. Finally, we found that inhibition of MyD88 restored GroEL-induced osteogenic and adipogenic differentiation and blocking JNK/MAPK or NF-κB signaling partly restored GroEL effects. CONCLUSION: In the current study, we revealed a potential interaction between bacteria and host cells by showing that GroEL directs the osteogenic and adipogenic differentiation of hPDLSCs by the involvement of JNK/MAPK and NF-κB signaling. This study provides evidence that bacterial products can influence the differentiation of stem cells and reveals potential effect of GroEL on the context of tissue regeneration.

SARS-CoV-2 detection in formalin-fixed paraffin-embedded tissue specimens from surgical resection of tongue squamous cell carcinoma.

In the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic, pathologists can be exposed to infection handling surgical specimens. Guidelines related to safety procedures in the laboratory have been released. However, there is a lack of studies performed on biopsy and surgical resection specimens. Here we report the detection of SARS-CoV-2 in formalin-fixed paraffin-embedded samples from surgical resection of tongue squamous cell carcinoma of a patient who developed COVID-19 postsurgery. RNA of SARS-CoV-2 strain was detected in the tumour and the normal submandibular gland samples using real-time PCR-based assay. No viral RNA was found in metastatic and reactive lymph nodes. We demonstrated that SARS-CoV-2 RNA can be detected in routine histopathological samples even before COVID-19 disease development. These findings may give important information on the possible sites of infection or virus reservoir, and highlight the necessity of proper handling and fixation before sample processing.

Myeloid sarcoma in the tongue.

Leukemic cells are rarely present in the oral cavity, and there are very few reports regarding such cases. However, we identified some reports of leukemic cells infiltrating tissues in the oral cavity, including gingival involvement. Recurrent painful oral ulcerations and prominent generalized periodontal destruction are the most common oral features of neutrophil disorders, and they may even be the initial symptoms of the disease. The ulcers may affect any part of the oral mucosa, including the tongue and palate. The objective of this report is to describe and discuss a case of myeloid sarcoma in the oral cavity of a 48-year-old male patient.

Surgical treatment of a rare bilateral synovial chondromatosis.

Synovial chondromatosis (SC) in the temporomandibular joint (TMJ) is an uncommon entity, mostly when the involvement is bilateral. The authors report a rare case of bilateral SC, with a follow-up of 13 months, and a literature review. A 60-year-old Caucasian woman, with the chief complaint of pain for 6 years in the bilateral pre-auricular region, had a progressive clacking and discomfort on the left side during mouth opening. The panoramic image was suggestive of SC. The bilateral lesion was surgically removed by direct access. Histopathological examination confirmed the clinical diagnosis of bilateral SC. This article shows the importance of a multidisciplinary approach for the early diagnosis and appropriate treatment. Also, it encourages the referral of such cases to professionals with a greater familiarity with this entity.

Chronic idiopathic neutropenia: A periodontist's diagnosis.

Our aim is to report the periodontal findings of a 10-year-old boy who visited the outpatient department of periodontology, with the chief complaint of swelling in the right cheek region for the last 2 months, increasing mobility of the teeth, and frequent bleeding from the gums. Since the age of 4 years, he suffered from recurrent febrile episodes, with boils and furuncles on the face. After several hematological and immunological investigations, he was diagnosed with chronic idiopathic neutropenia. He was prescribed a 150 µg subcutaneous injection of recombinant granulocyte colony-stimulating factor, once daily for 8 days. For reducing oral inflammation, he was advised an oral rinse of 15 ml of chlorhexidine gluconate (0.12%) twice daily and advised for a routine periodontal checkup, every 3-4 weeks for evaluation, maintenance, and avoiding any acute inflammatory flare-ups.

Variations of oral anatomy and common oral lesions

Abstract Several topics related to the oral cavity are briefly addressed in this article, from anatomical variations that, when recognized, avoid unnecessary investigations, to diseases that affect exclusively the mouth, mucocutaneous diseases, as well as oral manifestations of systemic diseases. A complete clinical examination comprises the examination of the mouth, and this approach facilitates clinical practice, shortening the path to diagnosis in the outpatient clinic as well as with in-hospital patients. The objective of this article is to encourage the examination of the oral cavity as a useful tool in medical practice, helping to recognize diseases in this location.

Prevalence of Oral Lesions Diagnosed at a Pathology Institute: A Four-year Analysis

Abstract Objective: To identify the most prevalent oral lesions based on reports from a pathology institute's reports and associations between malignant and oral potentially malignant disorders with patient's demographic variables and the anatomical location. Material and Methods: All 1,298 histopathological reports of oral lesions recorded in the database were reviewed. Demographic variables, anatomical location of the lesion, histopathological diagnosis of the lesions, and their biological behavior were analyzed. Results: Regarding the biological behavior of the identified lesions, benign lesions were predominant (70%), followed by lesions of undetermined behavior (14.3%), malignant lesions (14.2%), absence of histological alteration (1.2%), and finally, oral potentially malignant disorders (0.5%). The anatomical locations of the most prevalent oral lesions potentially malignant disorders and malignant were in the following structures of the oral cavity: gums, buccal mucosa, floor of the mouth and hard palate (p=49.2%), and tongue (p=48.7%). Conclusion: The probability of malignant and premalignant lesions was higher among males (PR= 4.21; 95% CI 2.08-6.22), the increase in age (PR = 1.06; 95% CI 1.05-1.08), and in the tongue region (PR = 5.48; 95% CI 1.67; 17.92). Identification of malignant and potentially malignant oral conditions is higher in older men and in tongue specimens.

Fibrous Dysplasia treated with virtual planning - report of two cases.

Fibrous dysplasia (FD) is part of a rare group of bone dysplasia. It exhibits benign behavior and can lead to osteolytic lesions, deformities, and fractures. The treatment is challenging, and accurate removal of the lesion is necessary to restore function and esthetics. Here we present two cases of FD where virtual planning with presurgical computed tomography (CT) was used for the production of a surgical guide for bone contouring. First, CT image reconstruction was performed to mirror the patient's original anatomy. Then, three surgical guides that determined the area and depth of bone wear were prepared and used in the relevant sequence during the actual surgeries, which were successfully performed in both patients. This technique is termed the template guide holes (TGH) technique. The findings from this report suggest that presurgical virtual planning and guide preparation allows direct and objective measurement of the level of bone wear and improves the functional and esthetic outcomes of surgery for FD. In particular, the TGH technique is safe and allows adequate preoperative surgical simulation, reduces the surgical duration, and increases the predictability of the final result.

Esthetic Management of Peripheral Giant Cell Granuloma Affecting a Dental Implant in the Esthetic Zone.

INTRODUCTION: A size increase of the gingiva can be a manifestation of gingival disease. While gingival lesions can affect periodontal tissue, only few reports described the association between peripheral giant-cell granuloma (PGCG) with dental implants and their clinical management. Their clinical relevance is of a great interest since some lesions may lead to extensive bone resorption, esthetic alterations, or even tooth/implant loss. To the author's best knowledge, no specific guidelines for the treatment of PGCG affecting dental implant in the esthetic zone have been reported. The aim of this paper was to report an extensive PGCG associated with a dental implant in the esthetic area and its surgical management. CASE PRESENTATION: A 39-year-old suffered from concussive trauma on her anterior maxillary dentition leading to an expanded, purplish soft tissue lesion surrounding the implant provisional crown on right central maxillary incisor. After complete lesion excision, the implant neck was carefully and gently debrided. Histological analysis confirmed the diagnosis of PGCG. The patient was then evaluated weekly for the first 3 weeks then monthly. No episodes of recurrence were identified. After 16 weeks of undisturbed healing, a soft tissue augmentation via tunnel technique was elected as the first treatment option to improve esthetics. Final implant restoration was delivered. Follow up at 22 months from biopsy excluded any recurrence of PGCG with stability of peri-implant soft tissues. CONCLUSION: Excisional biopsy, careful removal of irritating factors, monitoring, and connective tissue grafting can esthetically improve a malpositioned implant and showed no recurrence over a period of 22 months.