Maria Contaldo

Pilot study on reflectance confocal microscopy imaging of lichen planus: a real‐time, non‐invasive aid for clinical diagnosis

Background  Lichen planus (LP) represents a relatively common skin inflammatory entity included in the major group of interface dermatitis. In recent years, reflectance confocal microscopy has demonstrated to be a valuable tool for the ‘in vivo’ characterization of various skin diseases with cellular level resolution. No data are currently available that uses reflectance confocal microscopy to study LP.

Biological response of human bone marrow mesenchymal stem cells to fluoride‐modified titanium surfaces

OBJECTIVES The aim of the present study was to examine the behaviour of human bone marrow-derived mesenchymal stem cells (BM-MSC) to fluoride-modified grit-blasted (F-TiO) titanium surfaces compared with grit-blasted ones (TiO). MATERIAL AND METHODS Implant surfaces were analysed by atomic force microscopy (AFM) and scanning electron microscopy (SEM). BM-MSC were isolated from healthy donors and grown on the implant surfaces. Cell adhesion and proliferation, type I collagen (Col I) synthesis, osteoblastic differentiation (in terms of alkaline phosphatase activity, osteocalcin synthesis and extracellular matrix mineralization) were assessed. Furthermore, the ability to affect the osteoblastic/osteoclastic balance in terms of osteoprotegerin (OPG) and activator of nuclear factor κ B ligand (RANKL) ratio was investigated. RESULTS F-TiO surface showed higher S(a) values (P<0.05) and the presence of nano-scale structures at the AFM and SEM analysis. Comparable cell morphology and similar adhesion values on both surfaces were detected at early time, whereas higher proliferation values on F-TiO samples were observed at 7 and 10 days. Increased Col I and OPG levels for cells grown on F-TiO were found, whereas RANKL was not detectable in any of the conditioned media. BM-MSC showed a similar expression of early and late osteogenic markers on both TiO and F-TiO surfaces. CONCLUSIONS The results of the present study show that the chemical and micro/nano-scale modifications induced by fluoride treatment of TiO-grit blasted surfaces stimulate the proliferation and the extracellular matrix synthesis by BM-MSC, as well as the increase of OPG synthesis, thus preventing osteoclast activation and differentiation.

A Troubling Diagnosis of Verrucous Squamous Cell Carcinoma (“the Bad Kind” of Keratosis) and the Need of Clinical and Pathological Correlations: A Review of the Literature with a Case Report

Verrucous carcinoma (also known as Ackerman tumor) is an uncommon exophytic low-grade well-differentiated variant of squamous cell carcinoma. This neoplasm typically involves the oral cavity, larynx, genitalia, skin, and esophagus. It is well known for its locally aggressiveness and for its clinically slow-growing behaviour with minimal metastatic potential. Verrucous carcinoma of oral cavity is so closely aligned with the use of snuff and chewing tobacco that it has been called the “snuff dippers cancer”. Recent studies have proved the role of HPV. The typical clinical presentation of oral verrucous carcinoma has long been known, as its remarkably innocuous appearance and biological behaviour. In this work, we report a review of the scientific literature and describe a troublesome case of oral verrucous cancer.

The role of E-cadherin down-regulation in oral cancer: CDH1 gene expression and epigenetic blockage

BACKGROUND The prognosis of the oral squamous cell carcinoma (OSCC) patients remains very poor, mainly due to their high propensity to invade and metastasize. E-cadherin reduced expression occurs in the primary step of oral tumour progression and gene methylation is a mode by which the expression of this protein is regulated in cancers. In this perspective, we investigated E-cadherin gene (CDH1) promoter methylation status in OSCC and its correlation with Ecadherin protein expression, clinicopathological characteristics and patient outcome. METHODS Histologically proven OSCC and paired normal mucosa were analyzed for CDH1 promoter methylation status and E-cadherin protein expression by methylation-specific polymerase chain reaction and immunohistochemistry. Colocalization of E-cadherin with epidermal growth factor (EGF) receptor (EGFR) was evidenced by confocal microscopy and by immunoprecipitation analyses. RESULTS This study indicated E-cadherin protein down-regulation in OSCC associated with protein delocalization from membrane to cytoplasm. Low E-cadherin expression correlated to aggressive, poorly differentiated, high grade carcinomas and low patient survival. Moreover, protein down-regulation appeared to be due to E-cadherin mRNA downregulation and CDH1 promoter hypermethylation. In an in vitro model of OSCC the treatment with EGF caused internalization and co-localization of E-cadherin with EGFR and the addition of demethylating agents increased E-cadherin expression. CONCLUSION Low E-Cadherin expression is a negative prognostic factor of OSCC and is likely due to the hypermethylation of CDH1 promoter. The delocalization of E-cadherin from membrane to cytoplasm could be also due to the increased expression of EGFR in OSCC and the consequent increase of E-cadherin co-internalization with EGFR.

In Vivo Characterization of Healthy Oral Mucosa by Reflectance Confocal Microscopy: A Translational Research for Optical Biopsy

Abstract Purpose: To characterize in vivo microscopic features of healthy oral mucosa by noninvasive, real-time reflectance confocal microscopy (RCM). Material and Methods: One hundred healthy mucosal sites (38 lips, 24 cheeks, 10 gingivae, and 28 tongues) from 50 randomly selected volunteers were imaged by a commercially available in vivo RCM (Vivascope3000, Lucid, Rochester, NY). Morphological results were described and correlated with conventional histology. Results: Specific microscopic parameters (cellular morphology, anatomical structures features, architectural patterns) were identified by RCM for each mucosal site. Conclusions: RCM features reproducibility in different mucosal sites and subjects, which suggests that RCM can be an adjunct tool for clinical management in stomatology.

Oral lichen planus: a narrative review.

Oral Lichen Planus (OLP) is a common disease of unknown aetiology affecting oral mucosae by T-cell mediated chronic inflammation. OLP diagnosis is made by evaluating both clinical and histological criteria. Pharmacological treatment is useful in symptomatic cases. Life-long clinical follow-up is essential, due to low-risk of malignant transformation. In vivo Reflectance Confocal Microscopy (RCM) offers a real-time virtual biopsy of the being tissues and does not require surgical excision nor histopathological processing. RCM was used to capture OLP lesions in order to clinically differentiate them from other clinical entities.

Oral mucosa optical biopsy by a novel handheld fluorescent confocal microscope specifically developed: technologic improvements and future prospects.

OBJECTIVE This pilot study evaluated the baseline effectiveness of a novel handheld fluorescent confocal microscope (FCM) specifically developed for oral mucosa imaging and compared the results with the literature. STUDY DESIGN Four different oral sites (covering the mucosa of the lip and of the ventral tongue, the masticatory mucosa of the gingiva, and the specialized mucosa of the dorsal tongue) in 6 healthy nonsmokers were imaged by an FCM made up of a confocal fiberoptic probe ergonomically designed for in vivo oral examination, using light at the wavelength of 457 nm able to excite the fluorophore acriflavine hydrochloride, topically administered. In total, 24 mucosal areas were examined. RESULTS The FCM was able to distinctly define epithelial cells, bacterial plaque, and inflammatory cells and to image submucosal structures by detecting their intrinsic fluorescence. CONCLUSIONS When compared with other devices, this FCM allowed the user to image each oral site at higher magnification, thus resulting in a clearer view.

In Vivo Microstructural Analysis of Enamel in Permanent and Deciduous Teeth

Abstract Confocal microscope was used to analyze human enamel from 10 deciduous and 10 permanent teeth. Optically sectioned images were obtained. A more intense autofluorescence was found in primary teeth. This finding might be due to the greater presence of organic substances in deciduous enamel. The mean prism diameter measurement in permanent teeth enamel was 3.150 µm and 2.602 µm in deciduous teeth. The mean prism diameter in deciduous teeth was statistically least. The results indicate that a confocal microscope may be of help in analyzing and defining the microscopic features of human enamel.

Non-invasive in vivo visualization of enamel defects by reflectance confocal microscopy (RCM)

The enamel defects (EDs) may present with a variety of clinical manifestations with increasing severity from the sole appearance of pale discoloration to remarkable structural alterations. EDs are responsible for higher caries receptivity. In vivo reflectance confocal microscopy (RCM) allows to image in vivo at microscopic resolution of the dental surface, thus avoiding the tooth extraction and the sample preparation because of its ability to optically scan living tissues along their depth. Aim of this study is the in vivo assessment at microscopic resolution of dental surfaces affected by EDs without resorting to invasive methods such as teeth extractions, to define histological findings occurring in chromatic and/or structural EDs. For the purpose, 15 children, referring at the Dental Clinic of the Second University of Naples, affected by several degrees of EDs, were enrolled and underwent in vivo RCM imaging to microscopically define the ED confocal features using a commercially available hand-held reflectance confocal microscope with neither injuries nor discomfort. Totally, 29 teeth were imaged. Results demonstrated images good in quality and the capability to detect EDs such as unevenness, grooves, and lack of mineralization according to their clinical degree of disarray. The present in vivo microscopic study on EDs allowed to highlight structural changes in dental enamel at microscopic resolution in real-time and in a non-invasive way, with no need for extraction or processing the samples. Further experiments could define the responsiveness to remineralizing procedures as therapeutic treatments.

Prognostic implications of node metastatic features in OSCC: a retrospective study on 121 neck dissections

Lymph node metastases are responsible for shorter survival in oral squamous cell carcinoma (OSCC). The aim of the present study was to assess the node metastasis frequency and survival according to the node metastasis features in 121 neck dissections (NDs) performed for OSCC, identifying evidence-based correlations and contrasts with previous literature. The retrospective study involved 121 patients affected by OSCC who had undergone modified radical ND (MRND) for therapeutic, elective reasons or after intraoperative positivity to metastasis of sentinel lymph nodes (SLN+). Node metastasis frequency and behaviour (typical vs. atypical) and their number and distribution according to pre-surgical cTNM cancer staging were considered and overall survival Kaplan-Meier curves were calculated for each group in order to compare mortality according to ND type (elective, therapeutic, after SLN+), lymph node metastatic pattern (typical or atypical), size (micrometastasis vs. macrometastasis) and number. Results showed statistically significant different overall survival according to pre-surgical staging, number of lymph nodes harvested and intent to surgery. Sentinel lymph node resulted in the sole positive node affected by metastasis in small cT1- cT2/cN0 OSCC and an ND subsequent to its positivity during intraoperative assessment may be considered an overtreatment.