ALDH1 immunoexpression in epithelial and stromal cells of oral lichen planus and lesions with lichenoid inflammatory infiltrate

Background: Oral Lichen Planus is a potential malignant disorder and shares clinical and histopathological features with other similar lesions. ALDH1 is a specific biomarker for stem cells identification, however its role in stromal cells of immune inflammatory infiltrate has not been explored. The aim of this study was to investigate the ALDH1 immunoexpression in epithelial and stromal cells of Oral Lichen Planus and other lesions with lichenoid inflammatory infiltrate. Material and Methods: 64 samples of Oral Lichen Planus, Oral Lichenoid Lesions, Oral Leukoplakia and Unspecific Chronic Inflammation were included. ALDH1 was evaluated in both epithelium and stromal cells. ALDH1+ cells ≥ 5% were considered positive in epithelium. Stromal cells were evaluated semi quantitatively. Fields were ranked in scores, according to criteria: 1 (0 to 10%); 2 (11 to 50%) and 3 (>50%). The mean value of the sum of the fields was the final score. Statistical differences among groups were investigated, considering p < 0.05. Results: ALDH1 expression in epithelium was low in all groups without difference among them. ALDH1+ cells in the lamina propria were higher for Lichen Planus [2.0], followed by Leukoplakia [1.3], Lichenoid lesions [1.2] and control [1.1] (p<0.05). Conclusions: ALDH1 immunoexpression in epithelium of lichenoid potential malignant disorders did not show a contributory tool, however ALDH1 in stromal cells of lichen planus might be involved in the complex process of immune regulation associated with the pathogenesis of this disease. (AU)

Microvascular density analysis and histological parameters of oral cancer progression.

OBJECTIVES: This study aimed to investigate the role of blood and lymphatic microvascular density in the progression of oral squamous cell carcinoma (OSCC). MATERIALS AND METHODS: The sample was composed of 54 cases of OSCC. The immunoexpression to anti-alpha-smooth muscle actin (α-SMA) and to anti-endoglin (CD105) was used to determine the microvessel density (MVD); anti-podoplanin (D2-40) was used to assess the lymphatic vessel density (LVD); vascular endothelial growth factor (VEGF) was evaluated in malignant cells. The histological differentiation, the worst pattern of invasion (WPOI), tumour thickness and tumour budding (TB) intensity were assessed using haematoxylin-eosin and anti-pan-cytokeratin (AE1/AE3). Patients' age and sex, TNM classification and follow-up time were collected from the medical records. RESULTS: MVD markers presented a similar pattern of expression in blood vessels. However, only α-SMA + MVD was significantly higher among women and in tumours ≤4 cm. LVD was lower in tumours with lymph node metastasis. Regarding the histological parameters, high TB intensity was associated with histological differentiation, advanced clinical stage, greater tumour thickness and reduced disease-free survival. No difference was found in VEGF. CONCLUSIONS: The decrease in OSCC LVD could be related to pathological node involvement, whereas high TB intensity could indicate OSCC progression and worse patient outcomes.

Microvascular density and tumor budding in oral squamous cell carcinoma

Background: Oral squamous cell carcinoma (OSCC) is the most prevalent malignant head and neck tumor, excluding the nonmelanoma skin cancer. Despite recent advances in the diagnosis and treatment, the disease's mortality rate is nonetheless high. The presence of isolated neoplastic cells or small clusters of up to four cells at the tumor's invasive front, named tumor budding, is associated with a worse prognosis in OSCC. Angiogenesis has also been recognized as a determining factor in the progression of malignancies and in the development of metastases. Several studies have investigated the assessment of microvascular density (MVD) as a potential prognostic factor in OSCC. This study aimed to evaluate, in OSCC, differences in MVD between tumors with high-intensity tumor budding and tumors with low-intensity or no tumor budding. In samples with high-intensity tumor budding, differences in MVD between the budding area and the area outside the budding were also evaluated. Moreover, the study assessed differences in MVD concerning clinicopathological characteristics such as sex, age, tobacco smoking, tumor location and tumor size. Material and methods: One hundred and fifty [150] samples of OSCC were subjected to immunohistochemistry to assess the intensity of tumor budding (by immunostaining for multi-cytokeratin) and MVD (by immunostaining for CD34 and CD105, independently). The data were treated using descriptive and analytical statistics. (AU)

Lymphatic Vascular Density, the Expression of Podoplanin and Tumor Budding in Oral Squamous Cell Carcinoma.

BACKGROUND: Notwithstanding recent advances in oral squamous cell carcinoma (OSCC) management, its mortality rate is still high. It is imperative to investigate new parameters that are complementary to clinical staging for OSCC to provide better prognostic insight. The presence of isolated neoplastic cells or small clusters of up to four cells at the tumor's invasive front, called tumor budding, is a morphological marker of OSCC with prognostic value. Increased lymphatic vascular density (LVD) and a high expression of podoplanin in neoplastic cells have also been associated with worse prognosis in OSCC. To investigate these markers in OSCC, we evaluated differences in LVD and the expression of podoplanin in neoplastic cells between tumors with high-intensity tumor budding versus low-intensity or no tumor budding. In the samples of high-intensity budding, differences in those parameters between the​​ budding area and the area outside the budding were also evaluated. Furthermore, the study assessed differences in LVD and in the expression of podoplanin in neoplastic cells concerning OSCC clinicopathological characteristics. METHODS: To those ends, we subjected 150 samples of OSCC to immunohistochemistry to evaluate the intensity of tumor budding (via multi-cytokeratin immunostaining). Moreover, the 150 samples of OSCC and 15 specimens of normal oral mucosa (used as a control) were employed to assess LVD and the expression of podoplanin (in neoplastic cells of OSCC and in the lining epithelium of normal oral mucosa), both via podoplanin immunostaining. Data were processed into descriptive and analytical statistics. RESULTS: No differences were observed neither in the LVD nor in the expression of podoplanin in neoplastic cells concerning sex, age, tobacco smoking, tumor location and tumor size. The LVD was greater in OSCC and in tumors with high-intensity budding than in normal mucosa but did not differ between normal mucosa and tumors with low-intensity or no tumor budding. The data analyses also revealed that LVD was greater in tumors with high-intensity tumor budding than in tumors with low-intensity or no budding and showed no difference in LVD between the budding area and the area outside the budding. When compared to the lining epithelium of the normal mucosa, the expression of podoplanin was greater in neoplastic cells of OSCC, tumors with high-intensity budding and tumors with low-intensity or no tumor budding. The expression of podoplanin in neoplastic cells was also greater in tumors with high-intensity budding and, within those tumors, greater in the budding area than in the area outside de budding. CONCLUSION: Those findings support the hypothesis that tumor budding is a biological phenomenon associated with the progression and biological behavior of OSCC.

Mechanical Properties of Mini-Implants Used in Extra-Radicular Anchorage

ABSTRACT Objective: To evaluate the mechanical properties of mini-implants (MIs) manufactured from stainless steel and compare them with conventional titanium-aluminum-vanadium alloy MIs. Material and Methods: The following groups were formed: G1 (n=24), 8×1.5 mm steel MIs; G2 (n=24), 12×2.0 mm steel MIs; and G3 (n=24), 10×1.5 mm titanium MIs. The 72 MIs were inserted in the infra zygomatic crest region of the maxilla and retromolar trigone in the jaw of 10 pigs. Pull-out, insertion torque, fracture and percussion tests were performed in order to measure the tensile strength, primary stability and fracture strength of MIs. A digital torque gauge was used to measure insertion and fracture torque, a universal mechanical testing machine was used for pull-out testing and a periotest device was used to measure the micromovement of MIs. For morphological and MI component evaluation, scanning electron microscopy (SEM) was performed. D'Agostino & Pearson, Kruskal-Wallis, and Dunn post-hoc and normality tests were used. Results: G2 insertion and fracture torques were significantly higher than G1 and G3 insertion and fracture torques (p<0.05). The pull-out and percussion tests presented similar values among the groups. SEM revealed that the fracture point was predominantly on the fourth thread for steel MIs (G1 and G2) and on the seventh thread for titanium-aluminum-vanadium MIs (G3). Conclusion: The mechanical properties of stainless steel MIs are superior to those of titanium-aluminum-vanadium alloy MIs.

Endodontic cement penetration after removal of calcium hydroxide dressing using XP-endo finisher.

We aimed to evaluate the penetration of endodontic cement following the removal of calcium hydroxide (Ca(OH)2) dressing using the XP-endo Finisher in association with different irrigating solutions. Sixty premolars were instrumented and applied with a Ca(OH)2 dressing. To remove Ca(OH)2, the teeth were divided into six groups, each with a different volume of sodium hypochlorite (NaOCl) and ethylenediaminetetraacetic acid (EDTA), as well as solution stirring time with XP-endo Finisher (0, 30, and 60 sec). Root canals were filled using the lateral condensation technique. Fluorescein dye was added to the cement for microscopic laser scanning analysis. In the generated images, linear measurements were taken in micrometers, and their averages were calculated. To analyze the perimeter penetration ratio of the cement, the total perimeter of the canal and the segment of the total perimeter of the canal where the endodontic cement penetrated into the dentinal tubules were measured in micrometers. We found that using an XP-endo Finisher in irrigation was more effective than using a needle and syringe during the extension and penetration of endodontic cement. Shaking with XP-endo Finisher with 17% EDTA increased the extent and perimeter of the penetration of the endodontic cement into the dentinal tubules. However, using the XP-endo Finisher with EDTA only was more efficient than using the instrument interchangeably in NaOCl and EDTA. Although XP-endo Finisher contributes to the removal of Ca(OH)2, none of the protocols or instruments used removed all Ca(OH)2 from the root system.

Management of orofacial lesions with antimicrobial photodynamic therapy and photobiomodulation protocols in patients with COVID-19: A multicenter case series.

Several oral lesions related to COVID-19 have been described in the scientific literature. The COVID-19 pandemic highlighs importance of supportive protocols, which can reduce the inflammation and aid in tissue repair in severe cases. Photobiomodulation therapy (PBMT) alone or in combination with antimicrobial photodynamic therapy (aPDT) can be used to manage orofacial lesions in confirmed cases of COVID-19. Here, we sought to describe the clinical presentation and specificities of three cases in which aPDT and PBMT were used to manage orofacial lesions in patients with COVID-19. The laser protocols were effective with improvement of the orofacial lesions within a few days.

Cartilaginous choristoma on the lateral surface of the tongue: a case report.

A cartilaginous choristoma is a nonneoplastic nodular growth of histologically normal cartilage in an abnormal site. This report describes a case of a cartilaginous choristoma on the lateral surface of the tongue in a 65-year-old woman. The entire lesion was excised, and histologic examination revealed mature cartilaginous tissue surrounded by dense connective tissue. Choristomas are rare findings in the oral cavity, easily confused with proliferative processes or soft tissue neoplasms. Nevertheless, choristomas may be part of the differential diagnosis for lesions similar to the one described in this case report.

Acquired oral syphilis: A multicenter study of 339 patients from South America.

OBJECTIVE: To report the clinicopathologic features of acquired oral syphilis cases in South American countries. MATERIALS AND METHODS: Clinical data were retrospectively collected from the records of 18 oral diagnostic services in Argentina, Brazil, Chile, Colombia, Venezuela, Uruguay, and Peru. Serologies of nontreponemal and treponemal tests were used for diagnosis. RESULTS: The series comprised 339 cases of acquired oral syphilis. Secondary syphilis ranked as the most common stage (86.7%). Lesions were more frequent among males (58.0%) and young adults with a mean age of 33.3 years. Individuals aged 20-29 years were most affected (35.3%). The most commonly involved sites were the tongue (31.6%), lip/labial commissure (25.1%), and hard/soft palate (20.4%). Clinically, acquired oral syphilis usually presented as mucous patches (28.4%), papules (25.7%), and ulcers (18.1%). Skin manifestations occurred in 67.7% of individuals, while lymphadenopathy and fever were observed in 61.3% and 11.6% of all subjects, respectively. Most patients were treated with the benzathine penicillin G antibiotic. CONCLUSION: This report validates the spread of acquired oral syphilis infection among young adults in South America. Our directives include accessible diagnostic tools for proper disease screening, surveillance, and counselling of affected individuals, especially in low- and middle-income countries.

Endodontic cement penetration after removal of calcium hydroxide dressing using XP-endo finisher

Abstract: We aimed to evaluate the penetration of endodontic cement following the removal of calcium hydroxide (Ca(OH)2) dressing using the XP-endo Finisher in association with different irrigating solutions. Sixty premolars were instrumented and applied with a Ca(OH)2 dressing. To remove Ca(OH)2, the teeth were divided into six groups, each with a different volume of sodium hypochlorite (NaOCl) and ethylenediaminetetraacetic acid (EDTA), as well as solution stirring time with XP-endo Finisher (0, 30, and 60 sec). Root canals were filled using the lateral condensation technique. Fluorescein dye was added to the cement for microscopic laser scanning analysis. In the generated images, linear measurements were taken in micrometers, and their averages were calculated. To analyze the perimeter penetration ratio of the cement, the total perimeter of the canal and the segment of the total perimeter of the canal where the endodontic cement penetrated into the dentinal tubules were measured in micrometers. We found that using an XP-endo Finisher in irrigation was more effective than using a needle and syringe during the extension and penetration of endodontic cement. Shaking with XP-endo Finisher with 17% EDTA increased the extent and perimeter of the penetration of the endodontic cement into the dentinal tubules. However, using the XP-endo Finisher with EDTA only was more efficient than using the instrument interchangeably in NaOCl and EDTA. Although XP-endo Finisher contributes to the removal of Ca(OH)2, none of the protocols or instruments used removed all Ca(OH)2 from the root system.

Push-out bond strength and marginal adaptation of apical plugs with bioactive endodontic cements in simulated immature teeth.

OBJECTIVES: This study evaluates the bond strength and marginal adaptation of mineral trioxide aggregate (MTA) Repair HP and Biodentine used as apical plugs; MTA was used as reference material for comparison. MATERIALS AND METHODS: A total of 30 single-rooted teeth with standardized, artificially created open apices were randomly divided into 3 groups (n = 10 per group), according to the material used to form 6-mm-thick apical plugs: group 1 (MTA Repair HP); group 2 (Biodentine); and group 3 (white MTA). Subsequently, the specimens were transversely sectioned to obtain 2 (cervical and apical) 2.5-mm-thick slices per root. Epoxy resin replicas were observed under a scanning electron microscope to measure the gap size at the material/dentin interface (the largest and smaller gaps were recorded for each replica). The bond strength of the investigated materials to dentin was determined using the push-out test. The variable bond strengths and gap sizes were evaluated independently at the apical and cervical root dentin slices. Data were analyzed using descriptive and analytic statistics. RESULTS: The comparison between the groups regarding the variables' bond strengths and gap sizes showed no statistical difference (p > 0.05) except for a single difference in the smallest gap at the cervical root dentin slice, which was higher in group 3 than in group 1 (p < 0.05). CONCLUSIONS: The bond strength and marginal adaptation to root canal walls of MTA HP and Biodentine cement were comparable to white MTA.

Malignant lesion of clear cells: A diagnostic challenge of an unusual presentation mimicking a benign lesion.

In the oral cavity, a broad spectrum of differential diagnostics includes lesions composed of clear cells. Under this umbrella, Clear Cell Odontogenic Carcinoma (CCOC) and Hyalinizing Clear Cell Carcinoma (HCCC) are rare malignancies that arise from different histological origins. However, the histology is similar; therefore, differentiation of CCOC and HCCC lesions is difficult and, in some cases, impossible to distinguish morphologically and immunohistochemically. Herein, we report an unusual presentation of a well-defined nodular lesion affecting the maxillary gingiva in a 19-year-old female, which presented clinically as a benign or reaction etiology lesion. Microscopic evaluation showed a tumor composed of cell sheets with clear cytoplasm, separated by septa of fibrous tissue and invading the connective tissue. Tumor cells were positive for p63 and AE1/AE3 and negative for PAX8, smooth muscle actin (AML) and estrogen receptor. The diagnosis was carcinoma with a clear cell pattern, and it was not possible to distinguish between HCCC and CCOC. In this study, clinicopathologic, histologic, and immunohistochemistry features of CCOC and HCCC were discussed due to the challenging histological diagnosis. Radical surgical treatment and rehabilitation of the patient through graft and dental implants were performed. The patient is under follow-up with no signs of recurrence.

Two rare cases of oral metastasis arising from lung adenocarcinoma and esophageal carcinoma.

Metastasis to the oral cavity are rare, representing only 1% of all oral malignancies, and originate from various sites such as the breast, prostate, lung and kidney. Clinically, they can simulate reactive and inflammatory lesions common in the oral cavity, and the clinical and microscopic diagnosis of these metastasis is a challenge. In this article, we report two new cases of esophageal and lung metastasis to oral tissues, highlighting their clinical characteristics and the process of diagnostic elucidation. We emphasize the importance for clinicians to consider the possibility of metastatic lesions in the oral cavity in patients previously diagnosed with malignant lesions in distant tissues and organs. Key words:Diagnosis, esophageal squamous cell carcinoma, adenocarcinoma of lung, oral cavity, metastasis.

Can morphological features evaluated in oral cancer biopsies influence in decision-making? A preliminary study.

Tumor budding (TB) is a promising prognostic marker in many cancers including oral squamous cell carcinoma. The evaluation of TB in preoperative diagnostic biopsies has been proven be possible; therefore, the association of TB with other morphological features can represent an important aid in the previous treatment decision. This study aims to evaluate TB in oral squamous cell carcinoma (OSCC) biopsies, assessing its association with other morphological characteristics of the sample. A total of 56 cases of OSCC were investigated. In hematoxylin and eosin-stained slides, morphological features including histopathological grading and mode of invasion were evaluated in the deep invasive front. Moreover, immunohistochemistry was performed with anti-multi-cytokeratin antibody helping in the identification of TB, which was graded as low-intensity or no TB and high-intensity TB. Descriptive and bivariate analyses were performed, and the level of significance was set at 5%. The tongue was the most-affected site with 29 (51.7 %) tumors. The predominant mode of invasion (27-48.2 %) was by groups of neoplastic cells without clear boundaries. Of the cases investigated, 37 (66.1 %) were high-intensity TB, which was associated with the mode of invasion of the tumors (p < 0.05). All cases with the worst mode of invasion showed high-intensity TB. Preliminary results showed the potential of morphological features, such as TB and mode of invasion, evaluated in diagnostic specimens of OSCC, aiding in the treatment decision to select patients who could benefit from more-aggressive treatments.

Resinous adhesive systems differentially affect the expression of cytokines by human monocytes stimulated or not with Streptococcus mutans in vitro.

OBJECTIVES: The polymerization of adhesive systems is incomplete and the residual monomers that have been released have a cytotoxic capacity. Some teeth develop into pulp necrosis after composite resin restorations. Considering frequent pulpal inflammation in response to cariogenic bacteria, substances released from the patches could affect the cells of the inflammatory infiltrate and interfere with the mechanisms of defense against microorganisms and protection of pulpal tissue. The aim of this study was to evaluate the effect of substances released by different resinous adhesive systems on cell viability and cytokine expression by human monocytes stimulated in vitro with Streptococcus mutans. DESIGN: Peripheral blood mononuclear cells from 10 healthy subjects were stimulated with S. mutans and then incubated with supernatants obtained from the Single Bond Universal (SBU) or Clearfil SE Bond (CSEB) adhesive systems for eight hours. Staining with Annexin V and 7AAD for analysis of apoptosis were performed and detection of monocytes expressing cytokines IL-1α, IL-6, IL-8, IL-10, IL-12 and TNF-α were performed by flow cytometry. RESULTS: No treatment significantly affected apoptosis in monocytes. SBU supernatant increased the frequency of monocytes expressing IL-8 and decreased the monocytes expressing IL-10. Considering S. mutans-stimulated cells, while SBU increased the frequency of IL-8+ monocytes, CSEB reduced the frequency of IL-6 and TNF-α positive monocytes. CONCLUSIONS: Products released from SBU seem to induce proinflammatory effects on monocytes while those from CSEB show an anti-inflammatory outcome. These effects may interfere in the control of cytokine-mediated immunoinflammatory pulp reactions, both in the presence and absence of stimulation by cariogenic bacteria.