The Evolution of the Cephalometric Superimposition Techniques from the Beginning to the Digital Era: A Brief Descriptive Review.

Superimposition of craniofacial structures from radiographic examination has been always used for assessing changes in the maxilla-mandibular complexes, especially for the evaluation of potential changes occurring during growth as well as after orthodontic treatment and/or maxillofacial surgery. However, the availability of cone beam computed tomography (CBCT) and the recent advancement in 3D imaging have allowed the development of specific techniques for the registration and superimposition of virtual three-dimensional anatomical structures, improving the diagnosis and treatment plan strategies. In the present paper, it will be discussed the evolution of superimposition techniques from the beginning (2D) to the newest 3D approach, describing the most used methods and their main advantages and disadvantages, focusing primarily on accuracy and reproducibility of each technique.

Autophagy analysis in oral carcinogenesis.

OBJECTIVE: The aim of this study was to evaluate the levels of autophagy in oral leukoplakia and squamous cell carcinoma and to correlate with clinical pathological features, as well as, the evolution of these lesions. METHODOLOGY: 7 Normal oral mucosa, 51 oral leukoplakias, and 120 oral squamous cell carcinomas (OSCC) were included in the study. Histological sections of the mucosa and leukoplakias were evaluated throughout their length, while the carcinomas were evaluated using Tissue Microarray. After the immunohistochemical technique, LC3-II positive cells were quantified in the different epithelial layers of the mucosa and leukoplakias and in the microarrays of the squamous cell carcinomas. The correlation between positive cells with the different clinical-pathological variables and with the evolution of the lesions was tested using the t test, ANOVA, and Kaplan-Meier survival analysis. RESULTS: We observed increased levels of autophagy in the oral squamous cell carcinomas (p<0.001) in relation to the other groups, but without any association with poorer evolution or survival of these patients. Among the leukoplakias, we observed a higher percentage of positive cells in the intermediate layer of the dysplastic leukoplakias (p=0.0319) and in the basal layer of lesions with poorer evolution (p=0.0133). CONCLUSION: The levels of autophagy increased during the process of oral carcinogenesis and are correlated with poorer behavior of the leukoplakias.

Lubricin in synovial fluid of mild and severe temporomandibular joint internal derangements.

BACKGROUND: To understand the molecular basis of temporomandibular joint (TMJ) pathologies, we aimed to investigate the lubricin levels in the TMJ synovial fluid (SF) of patients with mild to severe internal derangements (IDs). MATERIAL AND METHODS: A total, 34 joints were the study group. Only patients, with a Wilkes stage of III, IV and V were included, in this sample. Control group consisted of SF from eight joints, from patients undergoing to orthognatic surgery. Concentrations of lubricin in the SF from both samples were measured using ELISA system. RESULTS: The mean lubricin concentration was 7.029 ± 0.21 µg/mL in stage III patients; 5.64 ± 0.10 µg/mL in stage IV patients, and 4.78 ± 0.11 µg/mL in stage V patients. The lubricin levels from stage IV and stage V patients differed significantly (P ≤ 0.001) from those of control subjects. Lubricin levels were inversely correlated with age and to VAS score. CONCLUSIONS: The results of this cross-sectional study highlight the relationship between disease severity and the levels of lubricin in TMJ SF. Our findings suggest that novel biotherapeutic approaches, including the administration of recombinant lubricin in the joint cavity, for the treatment of TMJ diseases can be developed.

Impacted maxillary canines and root resorption of adjacent teeth: A retrospective observational study.

BACKGROUND: The prevalence of impacted maxillary canine is reported to be between 1% and 3%. The lack of monitoring and the delay in the treatment of the impacted canine can cause different complications such as: displacement of adjacent teeth, loss of vitality of neighbouring teeth, shortening of the dental arch, follicular cysts, canine ankylosis, recurrent infections, recurrent pain, internal resorption of the canine and the adjacent teeth, external resorption of the canine and the adjacent teeth, combination of these factors. An appropriate diagnosis, accurate predictive analysis and early intervention are likely to prevent such undesirable effects. The objective is to evaluate, by means of a retrospective observational study, the possibility of carrying out a predictive analysis of root resorption adjacent to the impacted canines by means of orthopantomographs, so as to limit the prescription of additional 3D radiography. MATERIAL AND METHODS: 120 subjects with unilateral or bilateral maxillary impacted canine were examined and 50 patients with 69 impacted maxillary canine (22 male, 28 female; mean age: 11.7 years) satisfied the inclusion criteria of the study. These patients were subjected to a basic clinical and radiographic investigation (orthopantomographs and computerized tomography). All panoramic films were viewed under standardized conditions for the evaluation of two main variables: maxillary canine angulations (a, b, g angles) and the overlapping between the impacted teeth and the lateral incisor (Analysis of Lindauer). Binary logistic regression was used to estimate the likelihood of resorbed lateral incisors depending on sector location and angle measurements. RESULTS: Results indicated that b angle has the greatest influence on the prediction of root resorption (predictive value of b angle = 76%). If ß angle <18° and Lindauer = I, the probability of resorption is 0.06. CONCLUSIONS: Evaluation of b angle and superimposition lateral incisor/impacted canine analysed on orthopantomographs could be one of the evaluation criteria for prescribing second level examination (CT and CTCB) and for detecting root resorption of impacted maxillary canine adjacent teeth.

Adaptation and validation of an Italian version of the Prostate Cancer Specific Quality of Life Instrument (PROSQOLI).

OBJECTIVE: The Prostate Cancer Specific Quality of Life Instrument (PROSQOLI) is a measure of health-related quality of life (HRQoL) in advanced hormone-resistant prostate cancer. In this study, we aimed at performing a cross-cultural adaptation and validation of the Italian version of the PROSQOLI. PATIENTS AND METHODS: The original version of the PROSQOLI underwent several turnarounds of translations. A total of 472 patients treated with radical prostatectomy, radiotherapy or medical therapy were enrolled for the validation of the questionnaire. The PROSQOLI was administered together with the SF-12. Reliability indexes were calculated by using Cronbach alpha. To evaluate the validity of the construct, relationships between PROSQOLI and SF12 were assessed. The ANOVA test was used to evaluate the differences between groups of patients who had received different treatments. RESULTS: The reliability coefficient was 0.91. Item-to-total correlation indices were in most cases >0.70. The correlation between the scores of the PROSQOLI and those of the SF-12 questionnaire was high (r=0.8139, p<0.0001). The ANOVA test showed significant differences between groups (p<0.01) based on age, recurrence risk and treatment. CONCLUSIONS: The adaptation process showed that the PROSQOLI Italian version has high reliability and presents both convergent and discriminant validity. This version of the tool can be used to assess HRQoL in Italian men who underwent radical treatment for advanced prostate cancer.

Toll-like receptor 4 expression in the epithelium of inflammatory periapical lesions. An immunohistochemical study.

Toll-like receptors (TLR) are essential for the innate immune response against invading pathogens and have been described in immunocompetent cells of areas affected by periapical disease. Besides initiating the inflammatory response, they also directly regulate epithelial cell proliferation and survival in a variety of settings. This study evaluates the in situ expression of TLR4 in periapical granulomas (PG) and radicular cysts, focusing on the epithelial compartment. Twenty-one periapical cysts (PC) and 10 PG were analyzed; 7 dentigerous non-inflamed follicular cyst (DC) served as control. TLR4 expression was assessed by immunohistochemistry. TLR4 immunoreaction products were detected in the epithelium of all specimens, with a higher percentage of immunostained cells in PG. Although TLR4 overexpression was detected in both PG and PC, there were differences that seemed to be related to the nature of the lesion, since in PG all epithelial cells of strands, islands and trabeculae were strongly immunoreactive for TLR4, whereas in PC only some areas of the basal and suprabasal epithelial layers were immunostained. This staining pattern is consistent with the action of TLR4: in PG it could promote formation of epithelial cell rests of Malassez and in epithelial strands and islands the enhancement of cell survival, proliferation and migration, whereas in PC TLR4 could protect the lining epithelium from extensive apoptosis. These findings go some way towards answering the intriguing question of why many epithelial strands or islands in PG and the lining epithelium of apical cysts regress after non-surgical endodontic therapy, and suggest that TLR4 plays a key role in the pathobiology of the inflammatory process related to periapical disease.

Differential expression of TLR3 and TLR4 in keratocystic odontogenic tumor (KCOT): A comparative immunohistochemical study in primary, recurrent, and nevoid basal cell carcinoma syndrome (NBCCS)--associated lesions.

BACKGROUND: Toll-like receptors (TLRs) play an essential role in the activation of innate immunity and they can promote cancer cell survival and tumor progression. It has been claimed that TLRs can somehow predict the clinical behavior in oral squamous cell carcinoma (OSCCs). AIM: To elucidate the molecular basis underlying keratocystic odontogenic tumor (KOCTs) aggressive behavior and recurrence we carried out this immunohistochemical study on TLR3 and TLR4 expression in sporadic primary KCOTs (sp-KCOTs), sporadic recurrent KCOTs (sp-KCOTs), and NBCCS-associated KCOTs (NBCCS-KCOTs). METHOD: 40 cases of KOCTs removed from 23 men and 17 women were the sample. Paraffin-embedded blocks were processed for immunohistochemistry. Sections were incubated with TLR3 and TLR4 antibodies and immunoreactivity evaluated on a semi-quantitative score. RESULTS: Both TLR3 and TLR4 were expressed in KCOTs epithelium, although with a different extent. TLR3 was not expressed in sp-KCOTs and sr-KCOTs, but it showed a faint staining in NBCCS-KCOTs. On the other hand, both cytoplasmic and nuclear staining for TLR4 was detected in all the 3 types of lesions; however being significantly more expressed in sr-KCOT and NBCCS-KCOTs (p < 0.0001). Our results, demonstrated an association between TLR4, but not TLR3 expression to recurrence behavior of KCOTs. In fact, TLR4 was up-regulated in sr-KCOTs and NBCCS-KCOTs but not in sp-KCOTs. CONCLUSIONS: According these findings it seems conceivable to assume that the up-regulation of TLR4 in some KCOTs can be correlated somehow to their tendency recurrence.

Beta-catenin and survivin expression in keratocystic odontogenic tumor (KCOT). A comparative immunohistochemical study in primary, recurrent and nevoid basal cell carcinoma syndrome (NBCCS)-associated lesions.

AIM: To determine the epithelial expression of ß-catenin and survivin in sporadic (primary, and recurrent) and nevoid basal cell carcinoma syndrome (NBCCS) keratocystic odontogenic tumour (KCOT) in order to assess activation of the ß-catenin pathway and evidence of apoptotic inhibition, processes that may contribute to the known differences in their biological behaviour. MATERIALS AND METHODS: Sections from 40 cases of KCOT (19 sporadic/primary; 9 sporadic/recurrent and 12 NBCCS-associated) were immunohistochemically stained for ß-catenin and survivin. The extent and intensity of immunoreactivity within the lining epithelium was assessed, using semi-quantitative scales, independently by two pathologists who were blinded to the clinical-pathological data. Data were analysed using Kruskal-Wallis test and, for pair-wise comparisons, Mann-Whitney test with Bonferroni correction. RESULTS: All cystic epithelial linings stained for ß-catenin and survivin but there were differences in the pattern and intensity of staining among KCOT types. Sporadic primary KCOT showed weaker staining for ß-catenin (P=0.0003) and survivin (P<0.0048) that was restricted to the basal and para-basal layers only, compared to sporadic recurrent and NBCCS-associated KCOT, which showed expression throughout all epithelial layers. There were no differences in ß-catenin expression among recurrent and NBCCS-associated KCOT, whereas the intensity of survivin staining was higher in NBCCS-KCOT (P=0.0003). Nuclear staining for ß-catenin was found exclusively in recurrent (5/9 cases) and NBCCS-associated (4/12 cases) KCOT. CONCLUSION: The data demonstrate ß-catenin delocalization and survivin over-expression in recurrent sporadic and NBCCS-associated KCOT suggesting that these pathways related to apoptotic inhibition have a role in KCOT growth and recurrence.

Expression of TRAIL and its receptors DR5 and DcR2 in orthodontic tooth movement.

BACKGROUND: TRAIL is a transmembrane protein that induces apoptosis in various tissues including alveolar bone. Its in vitro expression can be activated by several methods, such as RANKL administration and cell scraping. Expression of TRAIL and its receptors DR5 and DcR2 was examined in osteoclast-like cells to analyze their effects on cell lifespan and to explore their role in orthodontic tooth movement. MATERIALS AND METHODS: Osteoclast-like cells were differentiated from a mouse hematopoietic cell line by stimulation with RANKL for 24 h (T1), 72 h (T2) or 5 days (T3); some cultures were then scraped. Immunostaining for TRAIL, DR5 and DcR2 was evaluated by immunocytochemistry and Western blot analysis in control and treated cells. RESULTS: Significantly greater TRAIL expression was found in treated osteoclast-like cells at T1 and T3 both on immunocytochemistry and Western blotting. TRAIL expression peaked at T1 and T3 in correspondence with DcR2 and DR5 maxima, respectively. CONCLUSIONS: These data may contribute to a better understanding of the mechanisms regulating tooth movement and to improve the accuracy of orthodontic treatments.

Possible role of apoptosis in the pathogenesis and clinical evolution of radicular cyst: an immunohistochemical study.

AIM: To investigate whether the apoptotic cascade is activated through the extrinsic pathway in epithelial lining and connective tissue of radicular cysts. METHODOLOGY: Fifteen radicular cysts were fixed in formalin, embedded in paraffin wax and processed for immunohistochemistry to evaluate the expression of polyclonal antibodies against Tumour necrosis factor-related apoptosis-inducing ligand (TRAIL), DR5 and caspase-3. Immunocomplexes were treated with the secondary antibodies and finally detected using the avidin-biotin-peroxidase complex. Immunoreactivity was visualized by development with 3,3'-diaminobenzidine. Data were analysed using the Mann-Whitney U-test; P < 0.05 was considered significant. RESULTS: The three antibodies were detected in connective tissue fibroblasts of all radicular cysts; TRAIL and DR5 immunoexpression was significantly greater (P < 0.05) compared with that of caspase-3. The three antibodies were also expressed in almost all epithelial layers and in endothelial cells of newly formed vessels. CONCLUSION: The involvement of apoptosis in the pathogenesis of radicular cysts, demonstrated by the immunoexpression patterns of TRAIL, DR5 and caspase-3 in lining epithelium and connective tissue, may explain their bland clinical aggressiveness and slow, benign evolution.

Lubricin is expressed in chondrocytes derived from osteoarthritic cartilage encapsulated in poly (ethylene glycol) diacrylate scaffold.

Osteoarthritis (OA) is characterized by degenerative changes within joints that involved quantitative and/or qualitative alterations of cartilage and synovial fluid lubricin, a mucinous glycoprotein secreted by synovial fibroblasts and chondrocytes. Modern therapeutic methods, including tissue-engineering techniques, have been used to treat mechanical damage of the articular cartilage but to date there is no specific and effective treatment. This study aimed at investigating lubricin immunohistochemical expression in cartilage explant from normal and OA patients and in cartilage constructions formed by Poly (ethylene glycol) (PEG) based hydrogels (PEG-DA) encapsulated OA chondrocytes. The expression levels of lubricin were studied by immunohistochemistry: i) in tissue explanted from OA and normal human cartilage; ii) in chondrocytes encapsulated in hydrogel PEGDA from OA and normal human cartilage. Moreover, immunocytochemical and western blot analysis were performed in monolayer cells from OA and normal cartilage. The results showed an increased expression of lubricin in explanted tissue and in monolayer cells from normal cartilage, and a decreased expression of lubricin in OA cartilage. The chondrocytes from OA cartilage after 5 weeks of culture in hydrogels (PEGDA) showed an increased expression of lubricin compared with the control cartilage. The present study demonstrated that OA chondrocytes encapsulated in PEGDA, grown in the scaffold and were able to restore lubricin biosynthesis. Thus our results suggest the possibility of applying autologous cell transplantation in conjunction with scaffold materials for repairing cartilage lesions in patients with OA to reduce at least the progression of the disease.

TRAIL, DR5 and caspase 3-dependent apoptosis in vessels of diseased human temporomandibular joint disc. An immunohistochemical study.

To evaluate the apoptosis involvement in the angiogenesis as a self-limiting process in patients with temporomandibular joint (TMJ) degenerated disc vessels, we assessed, by immunohistochemistry, the detection of TRAIL, its death receptor DR5 and caspase 3. TRAIL, its death receptor DR5 and caspase 3 expression were studied by immunohistochemistry in 15 TMJ discs displaced without reduction and in 4 unaffected discs. These apoptosis molecules were detected in the intima and media layers of newly formed vessels affected discs. In conclusion, vessels apoptosis activation in TMJ disc with ID could be regarded as a self-limiting process that try to leads to vessel regression; in this way an inhibition of angiogenic vessels may prove a key strategy in limiting pathological angiogenesis, by cutting off blood supply to tumors, or by reducing harmful inflammation.

MMP-13 expression in keratocyst odontogenic tumour associated with NBCCS and sporadic keratocysts.

OBJECTIVE: To investigate the matrix metalloproteinase (MMP)-13 expression in associated and non-nevoid basal cell carcinoma syndrome (NBCCS) Odontogenic Keratocysts (OCKs) in order to contribute to a better understanding of the differences in the growth pattern between them. MATERIALS AND METHODS: Thirty-nine paraffin-embedded blocks of OCKs, 26 sporadic OCKs and 11 NBCCS-associated KCOTs were studied by immunohistochemistry to evaluate MMP-13 expression both in epithelial and stromal layers. A semi-quantitative scale was used to evaluate immunostaining. Obtained data were compared between the two groups, using Fischer's exact test and the chi-square test. RESULTS: Only 13 of 26 sporadic OCKs showed a positive immunostaining, whilst 11 KCOTs resulted in positive labelling for MMP-13 expression. Moreover, syndromic cysts displayed a more intense and diffuse MMP-13 labelling of the stromal tissue. Instead, in non-syndromic forms, the staining pattern of MMP-13 in stromal tissue was completely absent. Fisher's exact test showed a statistically significant greater prevalence of KCOTs-immunolabelled cysts with respect to sporadic OCKs. CONCLUSIONS: Results from this study point out that the biological behaviour of these cysts could be related not only to the epithelial layer but also to stromal tissue in that... MMP-13 overexpression in stromal tissue of NBCCS-associated KCOTs could clarify the higher aggressiveness of these cysts.

Hepatocyte growth factor receptor, c-Met, in human embryo salivary glands. An immunohistochemical study.

Salivary gland morphogenesis involves complex, coordinated events that include epithelial-mesenchymal interactions. Mesenchymal-epithelial transition factor (c-Met) is the hepatocyte growth factor (HGF) receptor. The latter is a hepatotropic factor originally identified in rat serum and platelets. It is essential in fetal tissue development, where it regulates complex morphogenetic processes including extracellular matrix invasion, cell migration, cell polarization and tubulogenesis. The c-Met/HGF system is believed to participate in epithelial-mesenchymal interactions during development. Twelve human embryonic minor salivary glands were studied by immunohistochemistry to investigate the role of c-Met in human salivary gland development. Strong c-Met immunopositivity in the glands demonstrated that the molecule is involved in their development and suggested a role for the c-Met/HGF system in this process.

Simultaneous leiomyoma and controlateral smooth muscle hyperplasia of the epididymis: a case report.

We report a case of a 66-year-old man with a simultaneous leiomyoma and contralateral smooth muscle hyperplasia of the epididymis. The left nodule showed characteristics of a benign leiomyoma consisting in the homogeneous proliferation of smooth muscle with a typical pattern of interlacing fascicles of spindled smooth muscle cells showing no mitotic activity. The right nodule was made up of bundles of smooth muscle, growing in a perivascular and interstitial pattern, without the homogeneous aspect of contralateral leiomyomatous tumours. The latter finding suggested non-neoplastic hyperplasia of the smooth muscle fibres of the epididymis. At 6 months after surgery, ultrasound of the scrotum showed no evidence of recurrent lesions.

Preliminary results on selective light vaporization with the side-firing 980 nm diode laser in benign prostatic hyperplasia: an ejaculation sparing technique.

This study examined efficacy and safety of the 980 nm side-firing diode laser operating at a power of 100 W in patients with lower urinary tract symptoms associated with benign prostatic hyperplasia (BPH). Patients were selected for surgery on the basis of maximum urinary flow rate (Qmax) 100 ml in conjunction with an International Prostate Symptom Score (IPSS) >7. Laser therapy was conducted under spinal anaesthesia. Qmax, IPSS, prostate volume, PVR, IPSS-Quality of Life (QoL) questionnaire and ejaculatory function were measured at baseline and at 1, 3, 6 and 12 months post-laser therapy. A total of 52 consecutive patients aged 52-65 years with mean (s.d.) prostate volume of 45.14 (9.15) g were treated. All patients were able to leave the hospital at a mean (s.d.) of 5 (2) h after the procedure. Mean (s.d.) follow-up was 100 (75.49) days. A significant reduction in IPPS was reported at 1 month (P<0.0001) and this was maintained through to 6 months. Significant durable improvements in Qmax, PVR and IPSS-QoL score were also reported for all patients (P<0.0001). No severe complications were reported, including any cases of urinary incontinence or significant irritative symptoms. None of the patients complained of a worsening of erectile function. This early experience in a pilot study with the 980 nm diode laser seems promising in the short-term and long-term outcome is being monitored.

Chondrocyte-like apoptosis in temporomandibular joint disc internal derangement as a repair-limiting mechanism. An in vivo study.

Temporomandibular joint internal derangement (TMJ ID) is characterised by disc displacement and degenerative tissue changes involving an active cellular response, with cell phenotype transformation from fibroblast-like to fibrochondrocyte and, eventually, to chondrocyte-like, possibly as a response to abnormal loading. However, only small patches of chondral tissue are detected in TMJ discs with ID. We decided to explore the reasons for such incomplete tissue change, postulating an involvement of the apoptosis process. Twenty-one discs removed from 19 patients with TMJ ID were processed for TRAIL and DR5 immunohistochemical localisation, and subjected to the TUNEL assay. Overexpression of DR5 receptor and its ligand (TRAIL) in chondrocyte-like cells suggested activation of programmed cell death, as also demonstrated by TUNEL-positive cells. The data suggest a failed adaptive response to disc displacement through chondroid metaplasia. The apoptotic death of chondrocyte-like cells, which is at least partly regulated by TRAIL and its death receptor, appears to underpin the failed disc repair, eventually leading to its perforation.

Scleroderma-like disorders.

Fibrosing disorders comprise a wide spectrum of heterogeneous diseases characterized by sclerosis of the dermis, subcutis, and sometimes the underlying soft tissues and bone. The hallmark of this group of diseases is skin thickening as in systemic sclerosis with a different distribution pattern and for this reason they have also been referred to as "scleroderma-like" disorders. These diseases may have a different clinical course ranging from a benign disease with a localized cutaneous involvement, to a widespread, systemic, life-threatening disease. Some of them are associated with autoantibodies and/or autoimmune conditions. An accurate recognition of these scleroderma-like diseases is important for the institution of the most appropriate treatment.