Detection of HPV in oral leukoplakia by brushing and biopsy: prospective study in an Italian cohort Abstract Aim The aim of the present study was to evaluate the prevalence of HPV infection in oral leukoplakia, specifying the HPV genotypes eventually involved. We also compared the micro-biopsy and brushing HPV detecting efficacy. Materials and methods Consecutive patients with a presumptive diagnosis of oral leukoplakia were enrolled. Demographical, behavioral data (smoking, alcohol) and lesion features were recorded. Each patient underwent a brushing procedure, performed with a cytobrush rubbed on the lesion, and then a biopsy was performed. The brushing and micro-biopsy specimens were both analyzed with the HPV 28 Anyplex II Seegene RT-PCR. The prevalence of HPV infection was calculated considering the two methods’ outcomes separately and then combining both. Cohen’s k coefficient was used to assess the agreement between the two methods. Results Sixty-five patients were enrolled with a mean age of 60 years. The HPV infection prevalence was 17%, decreasing to 5% considering the brushing outcomes alone. The most frequently detected genotypes were 6 (12%), 11 (3%), 42 (3%), and 16 (3%). No statistically significant correlation was found between HPV infection and the variables analyzed, except for smoking and the type of mucosa (p < 0.05). The strength of agreement between cytobrush and micro-biopsy was “fair” (k = 0.384). Conclusions The present study showed a low prevalence of HPV infection in oral leukoplakia. The micro-biopsy appeared to be more reliable than brushing in detecting HPV DNA in oral leukoplakia, but the method invasiveness discourages its employ as a screening tool. The importance of HPV in the etiopathogenesis of oral potentially malignant lesions remains unclear; further studies are needed to establish the HPV role in oral leukoplakia. Clinical relevance HPV involvement in oral leukoplakia and an effective and appropriate detecting technique are still a debated issue. From this study, the restricted use of brushing did not appear sufficient to assess the presence of HPV infection with PCR techniques in samples obtained from oral leukoplakia.

A modified protocol of mandibular osteoradionecrosis induction in rats with external beam radiation therapy Abstract Objective To propose a modified protocol of mandibular osteoradionecrosis induction in rats with external beam radiation therapy. Material and methods A total of 45 male Wistar rats were used in this study. Firstly, 25 rats were divided into 5 groups (n = 5) according to the radiation dose protocol: without irradiation and irradiated with 15 Gy, 20 Gy, 25 Gy, or 30 Gy using a linear accelerator. Secondly, 15 other rats were divided into 3 groups (n = 5) according to the time of extraction of the three right mandibular molars: 7, 10, or 14 days after irradiation of 20 Gy. Lastly, dental extractions were performed in 5 other rats without irradiation (C-E10) for comparison with those of the group of dental extractions 10 days after irradiation (I-E10). Results The irradiated animals survived throughout the study period only at single doses of 15 Gy and 20 Gy. The suitable time for dental extractions after irradiation to induce mandibular osteoradionecrosis was defined as 10 days. Macroscopic evaluation of the right hemimandibles showed presence of osteoradionecrosis in I-E10 group and complete gingival healing in C-E10 group. Conclusion A single radiation dose of 20 Gy focused on head and neck region combined with the extraction of the three mandibular molars 10 days after irradiation constitutes a feasible protocol of mandibular osteoradionecrosis induction in rats with external beam radiation therapy. Clinical relevance Establishing a solid and widely available protocol of mandibular osteoradionecrosis induction is essential in the search for methods to prevent this complex disease.

Cytocompatibility, bioactivity potential, and ion release of three premixed calcium silicate-based sealers Abstract Objective Compositional modifications may alter the biological and physicochemical characteristics of calcium silicate-based sealers (CSBS) and, ultimately, their bioactivity. The main objective of this study was to evaluate the biological properties of three CSBS: EndoSequence BC Sealer, Ceraseal, and Endoseal mineral trioxide aggregate. Materials and methods Human periodontal ligament stem cells (hPDLSCs) were exposed to several eluates of CSBS. The ion release profile and pH were determined, and metabolic activity and cell migration were assessed using the MTT and wound healing assays. hPDLSCs were cultured in direct contact with the surface of each material, and cell morphology and attachment were analyzed by scanning electron microscopy (SEM). Bioactivity potential was assessed by RT-qPCR and mineralization assays. Statistical differences between biomaterials were assessed using one- or two-way ANOVA (α < 0.05). Results All materials showed an alkaline pH, although Endoseal exhibited a significantly higher pH compared with the other CSBS (p < 0.05). Ceraseal released significantly more Ca2+ (p < 0.05) than EndoSequence BC Sealer and Endoseal. Interestingly, Endoseal induced a significant reduction in cell viability and cell migration compared with the control (p < 0.001). Moreover, SEM showed abundant cells adhering to EndoSequence BC Sealer and Ceraseal surfaces, whereas very few round cells were detected on the surface of Endoseal. Finally, Ceraseal and EndoSequence induced ALP, CAP, and CEMP-1 expression and a significantly higher mineralization capacity than Endoseal (***p < 0.001). Conclusions The eluates from EndoSequence BC Sealer and Ceraseal displayed higher cell viability, cell attachment, cell migration rates, and ion release rates than Endoseal. Ceraseal and EndoSequence BC Sealer exhibited significantly more gene expression and mineralization capacity than Endoseal. Clinical relevance The results obtained in the present work suggest that EndoSequence BC Sealer and Ceraseal possess biological properties that make them suitable materials for root canal treatment.

Variation of human salivary alpha-amylase proteoforms in three stimulation models Abstract Objectives To evaluate the sAA proteoforms’ expression during different stimulation situations. Materials and methods This study evaluated the salivary alpha-amylase (sAA) proteoforms’ behavior by western blot (WB) analysis and high-resolution mass spectrometry (LC-MS/MS) in different situations that produce increases in sAA activity. For this purpose, six healthy women with a similar body mass index, age, and fit, underwent different sAA stimulation tests, such as acetic acid stimulation, psychological stress using the standardized Trier social stress test, and physical effort using the Cooper treadmill test. Results The three models showed an increase in sAA activity. The WB demonstrated seven common bands observed in the six women (band one at 59 kDa, two at 56 kDa, three at 48 kDa, four at 45 kDa, five at 41 kDa, six at 36 kDa, and seven at 14 kDa), in which sAA protein was identified. The individual WB analysis showed that band two, which corresponded to the native non-glycosylated sAA proteoform, had a higher increase after the three sAA stimulation inducers, and this band was also the only proteoform correlated with sAA activity (r = 0.56, P = 0.001). In addition, when the label-free quantification analysis was performed, the different proteoforms showed different responses depending on the type of stimulation. Conclusions This preliminary study showed that the diverse sAA proteoforms’ expression depends on the different stimulation models. Clinical relevance This study opens new perspectives and challenges for the use of the different alpha-amylase proteoforms as possible biomarkers in addition to the sAA activity.

Large versus small mandibular counterclockwise rotation during bimaxillary surgical correction of class II deformities—a retrospective CBCT study on skeletal stability Abstract Objectives Postsurgical skeletal relapse is a concern for class II deformities corrected with counterclockwise rotation of the occlusal plane. Therefore, the aim of this study was to compare the skeletal stability between large and small counterclockwise rotational advancement of the mandible in patients with skeletal class II deformity. Materials and methods This retrospective study included 50 adult patients with skeletal class II deformity corrected by Le Fort I setback and bilateral sagittal split osteotomy counterclockwise rotational advancement. Patients were divided into two groups, according to the amount of counterclockwise rotation: small rotation (n = 25) and large rotation (n = 25). Serial cone beam computed tomography scans were analyzed to identify skeletal and dental position from presurgery to at least 12 months postsurgery. Changes in the facial skeleton (maxilla and mandible) and teeth (central incisor and first molar) were determined for six skeletal and four dental landmarks by measures before treatment (T0) and 1 week postsurgery (T1), and from T1 to at least 12 months postsurgery (T2). Results A relapse was found both after large and small rotational advancement of the mandible (pogonion: 1.0 (2.4) mm and 1.4 (3.0) mm, respectively). The result was statistically significant (both p < 0.05) and was with less than 1.5 mm clinically acceptable. There were no between-group differences in the postsurgical horizontal and vertical mandibular stability. Conclusions The results suggest that counterclockwise rotational advancement of the mandible using bilateral sagittal split osteotomy is a clinically stable procedure. The amount of rotation does not affect the postsurgical stability of the mandible. Clinical relevance The findings help clinicians better understand the surgical and postsurgical changes of the skeleton and teeth after counterclockwise rotational advancement of the mandible for class II deformity.

Maxillofacial injuries in severely injured patients after road traffic accidents—a retrospective evaluation of the TraumaRegister DGU® 1993–2014 Abstract Objectives It was the aim of the study to analyse the prevalence of maxillofacial trauma (MFT) in severely injured patients after road traffic accidence (RTA) and to investigate associated factors. Materials and methods In a retrospective study, data from patients after RTA by the TraumaRegister DGU® from 1993 to 2014 were evaluated for demographical and injury characteristics. The predictor variable was mechanism of injury and the outcome variables were type of injury, severity and hospital resources utilization. Results During the investigation period, n = 62,196 patients were enclosed with a prevalence of maxillofacial injuries of 20.3% (MFT positive). The injury severity score of MFT-positive patients was higher than in the MTF-negative subgroup (27 ± 12.8 vs. 23.0 ± 12.7). If MFT positive, 39.8% show minor, 37.1% moderate, 21.5% serious and 1.6% severe maxillofacial injuries. Injuries of the midface occurred in 60.3% of MTF-positive patients. A relevant blood loss (> 20% of total blood volume) occurred in 1.9%. MFT-positive patients had a higher coincidence with cervical spine fractures (11.3% vs. 7.8%) and traumatic brain injuries (62.6% vs. 34.8%) than MFT-negative patients. There was a noticeable decrease in the incidence of facial injuries in car/truck drivers during the study period. Conclusions Every 5th patient after RTA shows a MFT and the whole trauma team must be aware that this indicates a high prevalence of traumatic brain and cervical spine injuries. Clinical relevance Even if sole injuries of the face are seldom life threatening, maxillofacial expertise in interdisciplinary trauma centres is strongly recommended.

Investigation of the relationship between sleep disorders and xerostomia Abstract Objectives To investigate the relationship between sleep disorders, morning hyposalivation, and subjective feeling of dry mouth. Materials and methods A cross-sectional, observational, clinical study was carried out in a homogenous population sample which consists of Greek male soldiers without any medical history. After the application of oral modified Schirmer test, the sample was divided into a study group (n = 63) (MST < 25 mm/3 min) and a control group (n = 110) (MST ≥ 25 mm/3 min). In order to assess daytime sleepiness, risk of obstructive sleep apnea (OSA), sleep quality, sleep bruxism (SB), and subjective feeling of dry mouth, all the participants filled in the following scales in Greek version: Epworth Sleepiness Scale (ESS), Pittsburgh Sleep Quality Index (PSQI), Berlin Questionnaire (BQ), a SB questionnaire, and Xerostomia Inventory (XI) respectively. In every subgroup that came of ESS, PSQI, BQ, and SB questionnaire scoring, subjective feeling of dry mouth was evaluated, based on XI values. Results Statistically significant difference (p < 0.001) through PSQI scores was found between the study and control group. In contrast, a statistically significant difference was not obtained for the scores of ESS (p = 0.293), BQ (p = 0.089), and SB questionnaire (p = 0.730). XI scores introduced statistically significant difference between the subgroups of PSQI (p < 0.001), BQ (p = 0.001), SB questionnaire (p = 0.004)  and statistically weak between the subgroups of ESS (p = 0.049). Conclusions This is the first research study so far suggesting that patients with morning hyposalivation exhibit poor sleep quality using an objective method. The present results have, also, shown that subjective feeling of dry mouth is related to excessive daytime sleepiness, poor sleep quality, high risk of obstructive sleep apnea, and sleep bruxism, but larger-scale studies are still needed. Clinical Relevance These findings should keep dentists aware of a possible association between xerostomia and sleep disorders and support larger-scale studies.

Incorporation of AuNP-PLL nanocomplexes in DPSC: a new tool for 3D analysis in pulp regeneration Abstract Objectives To assess the viability of dental pulp stem cells loaded with gold nanoparticles complexed with poly (l-lysine) (AuNP-PLL) and to track the cellular behavior in a 3D analysis by micro-CT. Materials and methods DPSC (dental pulp stem cells) were cultured and incorporated with AuNP-PLL (0.2 mg/ml) and assessed for cell viability (24 h, 48 h, and 72 h) using MTS assay. Apoptosis/cell death index and cell cycle were analyzed by propidium iodide. AuNP-PLL-RITC were used for observation in confocal microscopy and quantification of the incorporation rates. Cells were also suspended in agarose and analyzed three-dimensionally in μCT, assessing their radiopacity. Quantitative data (cell viability and apoptosis) were analyzed by t test (p < 0.05). Results AuNP-PLL labeling did not affect cellular viability in any of the periods analyzed nor interfered with the apoptosis index of DPSC. AuNP-PLL nanocomplexes were identified in the cytoplasm of cells by immunofluorescence, mainly in the perinuclear region. The observed incorporation rate was 98%. Micro-CT analysis has shown that incorporated cells are now visible using x-ray, with a clear increase in radiopacity when compared to the control group (non-incorporated cells). Conclusion These results indicate that it is possible to incorporate AuNP-PLL complex into DPSC and track the cells by using μCT; furthermore, this incorporation of 0.2 mg/ml of AuNP-PLL does not interfere in the DPSC basic behavior. Clinical relevance This methodology can be a useful tool for cellular labeling to observe cell behavior and their interaction with scaffolds in a 3D manner, opening an array of new approaches in regenerative endodontics.

Correction to: Sinus augmentation analysis of the gradient of graft consolidation: a split-mouth histomorphometric study Figure 1 was reused with permission from "Wiley", the publisher of a previous article by the same authors, DOI:10.1111/cid.12518.

Correction to: The fate of root canals obturated with Thermafil: 10-year data for patients treated in a master’s program The author names in the original version of this article were inadvertently interchange. Correct presentation of author names is reflected here.