Assessment of gingival translucency at the mandibular incisors with two different probing systems. A cross sectional study.

OBJECTIVES: Increasing evidence indicates that the thickness of periodontal soft tissues plays an important role in various clinical scenarios, thus pointing to the need of further clinical research in this area. Aim of the present study was to assess gingival thickness at the mandibular incisors by translucency judgement with two different probes and to validate if these methods are comparable and applicable as diagnostic tools. MATERIALS AND METHODS: A total of 200 participants were included; gingival tissue thickness was measured by judging probe translucency at both central mandibular incisors, mid-facially on the buccal aspect of each tooth using a standard periodontal probe and a set of color-coded probe, each with a different color at the tip, i.e. Colorvue Biotype Probe (CBP). Frequencies and relative frequencies were calculated for probe visibility. Agreement between the standard periodontal probe and the CBP was evaluated via the kappa statistic. RESULTS: When the periodontal probe was visible, the frequency of CBP being visible was very high. Kappa statistic for the agreement between the standard periodontal probe and the CBP was 0.198 (71.5% agreement; p-value < 0.001) for tooth 41 and 0.311 (74.0% agreement; p-value < 0.001) for tooth 31, indicating a positive association of the two methods. CONCLUSIONS: An agreement that reached 74% was estimated between the standard periodontal probe and the color-coded probe at central mandibular incisors.  CLINICAL RELEVANCE: In the context of the present study, the two methods of evaluating gingival thickness seem to produce comparable measurements with a substantial agreement. However, in the 1/4 of the cases, the visibility of the color-coded probe could not assist in the categorization of the gingival phenotype.

Accuracy of cone-beam computed tomography in imaging the components of the periodontal phenotype.

The components and dimensions of the periodontal and peri-implant phenotype have a high relevance in contemporary dental research and should be taken into consideration in the decision-making process in the management of a variety of clinical scenarios to optimize the outcomes of therapy. Various assessment methods for quantifying and classifying the phenotypical dimensions have emerged and developed in recent decades. Nevertheless, the use of cone-beam computed tomography (CBCT) scans remains the most commonly used approach worldwide. However, the accuracy to adequately imaging and measuring the dimensions of the hard and soft tissue components around teeth may represent a significant challenge in different clinical scenarios due to factors such as the age of the patient and motion during the scan, presence of metallic artifacts causing streaks and gray-value distortion, overlapping of soft tissue structures, machine performance, file processing, and small voxel size among others. These factors pose a particular challenge when tiny structures are under investigation, for example, the buccal/lingual bony or soft tissue layer of lower/upper incisors. Therefore, this review addresses the underlying technical information of the use of CBCT scans, and suggests some recommendations on the utilization of this method of assessment to optimally use it despite its' system-inherent limitations.

Long-term occlusal tooth wear at the onset of permanent dentition.

OBJECTIVES: This study quantified the long-term occlusal wear in the natural posterior teeth and the associations per tooth type within the dentition. METHODS: The sample included 70 orthodontically treated subjects (52 females and 18 males; median age, 14.3 years), followed for a 12.7-year period. They were consecutively selected with no tooth wear-related criteria. Post-treatment (T1) and follow-up dental casts (T2) were scanned and superimposed through three-dimensional methods. Occlusal wear volume of posterior teeth and tooth wear patterns were investigated through non-parametric statistics and analysis of covariance. RESULTS: There were no significant differences between contralateral teeth. The average occlusal wear per posterior tooth was 2.3 mm3, with 65.2% of teeth showing values greater than 1 mm3. Males, mandibular teeth, and first molars exhibited slightly greater wear levels than females (median, 2.57 and 2.21 mm3, respectively; p = 0.005), maxillary teeth, and first or second premolars, respectively. In all first premolars and in the mandibular second premolars, the buccal cusps were primarily affected with no other distinct patterns. There were weak to moderate correlations between tooth types, apart from certain strong correlations detected in males. CONCLUSIONS: Posterior tooth wear was highly prevalent after a 13-year period starting at the onset of permanent dentition. The detected patterns are in accordance with the concept of canine guidance occlusion that is transforming into group synergy through function. CLINICAL RELEVANCE: The widespread tooth wear occurrence and the high intra- and inter-individual variability underline the need for individual patient monitoring to identify high-risk patients at early stages.

Reflecting the human lip in vitro: Cleft lip skin and mucosa keratinocytes keep their identities.

OBJECTIVES: Cell models have shown great promise as tools for research, potentially providing intriguing alternatives to animal models. However, the original tissue characteristics must be maintained in culture, a fact that is often assumed, but seldom assessed. We aimed to follow the retention of the original tissue identities of cleft lip-derived skin and mucosa keratinocytes in vitro. METHODS: Cleft lip-derived keratinocytes were isolated from discarded tissue along the cleft margins during cheiloplasty. Cell identities were assessed by immunohistochemistry and quantitative real-time PCR for tissue-specific markers and compared with native lip tissue. Moreover, keratinocytes were regularly analyzed for the retention of the original tissue characteristics by the aforementioned methods as well as by differentiation assays. RESULTS: The various anatomical zones of the human lip could be distinguished using a panel of differentiation and functional-based markers. Using these markers, retention of the original tissue identities could be followed and confirmed in the corresponding primary keratinocytes in culture. CONCLUSIONS: Our findings promote patient-derived cells retaining their original identities as astonishing and clinically relevant in vitro tools. Such cells allow a better molecular understanding of various lip-associated pathologies as well as their modeling in vitro, including but not restricted to orofacial clefts.

Facial asymmetry and midsagittal plane definition in 3D: A bias-free, automated method.

Symmetry is a fundamental biological concept in all living organisms. It is related to a variety of physical and social traits ranging from genetic background integrity and developmental stability to the perception of physical appearance. Within this context, the study of human facial asymmetry carries a unique significance. Here, we validated an efficient method to assess 3D facial surface symmetry by best-fit approximating the original surface to its mirrored one. Following this step, the midsagittal plane of the face was automatically defined at the midpoints of the contralateral corresponding vertices of the superimposed models and colour coded distance maps were constructed. The method was tested by two operators using facial models of different surface size. The results show that the midsagittal plane definition was highly reproducible (maximum error < 0.1 mm or°) and remained robust for different extents of the facial surface model. The symmetry assessments were valid (differences between corresponding bilateral measurement areas < 0.1 mm), highly reproducible (error < 0.01 mm), and were modified by the extent of the initial surface model. The present landmark-free, automated method to assess facial asymmetry and define the midsagittal plane of the face is accurate, objective, easily applicable, comprehensible and cost effective.

Accuracy of facial skeletal surfaces segmented from CT and CBCT radiographs.

The accuracy of three-dimensional (3D) facial skeletal surface models derived from radiographic volumes has not been extensively investigated yet. For this, ten human dry skulls were scanned with two Cone Beam Computed Tomography (CBCT) units, a CT unit, and a highly accurate optical surface scanner that provided the true reference models. Water-filled head shells were used for soft tissue simulation during radiographic imaging. The 3D surface models that were repeatedly segmented from the radiographic volumes through a single-threshold approach were used for reproducibility testing. Additionally, they were compared to the true reference model for trueness measurement. Comparisons were performed through 3D surface approximation techniques, using an iterative closest point algorithm. Differences between surface models were assessed through the calculation of mean absolute distances (MAD) between corresponding surfaces and through visual inspection of facial surface colour-coded distance maps. There was very high reproducibility (approximately 0.07 mm) and trueness (0.12 mm on average, with deviations extending locally to 0.5 mm), and no difference between radiographic scanners or settings. The present findings establish the validity of lower radiation CBCT imaging protocols at a similar level to the conventional CT images, when 3D surface models are required for the assessment of facial morphology.

Synchrotron X-rays reveal the modes of Fe binding and trace metal storage in the brown algae Laminaria digitata and Ectocarpus siliculosus.

Iron is accumulated symplastically in kelp in a non-ferritin core that seems to be a general feature of brown algae. Microprobe studies show that Fe binding depends on tissue type. The sea is generally an iron-poor environment and brown algae were recognized in recent years for having a unique, ferritin-free iron storage system. Kelp (Laminaria digitata) and the filamentous brown alga Ectocarpus siliculosus were investigated using X-ray microprobe imaging and nanoprobe X-ray fluorescence tomography to explore the localization of iron, arsenic, strontium, and zinc, and micro-X-ray absorption near-edge structure (µXANES) to study Fe binding. Fe distribution in frozen hydrated environmental samples of both algae shows higher accumulation in the cortex with symplastic subcellular localization. This should be seen in the context of recent ultrastructural insight by cryofixation-freeze substitution that found a new type of cisternae that may have a storage function but differs from the apoplastic Fe accumulation found by conventional chemical fixation. Zn distribution co-localizes with Fe in E. siliculosus, whereas it is chiefly located in the L. digitata medulla, which is similar to As and Sr. Both As and Sr are mostly found at the cell wall of both algae. XANES spectra indicate that Fe in L. digitata is stored in a mineral non-ferritin core, due to the lack of ferritin-encoding genes. We show that the L. digitata cortex contains mostly a ferritin-like mineral, while the meristoderm may include an additional component.

Impact of fixed orthodontic appliances on blood count and high-sensitivity C-reactive protein levels: A prospective cohort study.

INTRODUCTION: The aim was to elucidate the magnitude of alterations in systemic blood counts in healthy patients during the first 14 days after fixed orthodontic appliance placement. METHODS: This prospective cohort study consecutively included 35 White Caucasian patients starting orthodontic treatment with fixed appliances. The mean age was 24.48 ± 6.68 years. All patients were physically and periodontally healthy. Blood samples were collected at 3 time points: (1) baseline (exactly before the placement of appliances), (2) 5 days after bonding, and (3) 14 days after baseline. Whole blood and erythrocyte sedimentation rates were analyzed in automated hematology and erythrocyte sedimentation rate analyzer. Serum high-sensitivity C-reactive protein levels were measured by the nephelometric method. Standardized sample handling and patient preparation procedures were adopted to reduce preanalytical variability. RESULTS: A total of 105 samples were analyzed. All clinical and orthodontic procedures were performed without complications or side effects during the study period. All laboratory procedures were performed per protocol. Significantly lower white blood cell counts were detected 5 days after bracket bonding, compared with baseline (P <0.05). Hemoglobin levels were lower at 14 days than baseline (P <0.05). No other significant shifts or alteration patterns were observed over time. CONCLUSIONS: Orthodontic fixed appliances led to a limited and transient change in white blood cell counts and hemoglobin levels during the first days after bracket placement. The fluctuation of high-sensitivity C-reactive protein levels was not significant, demonstrating a lack of association between systemic inflammation and orthodontic treatment.

Survival of conventionally bonded mandibular retainers with or without enamel sandblasting in orthodontic patients over a 12-month period. A single-centre, split-mouth randomized clinical trial.

BACKGROUND: The aim of this trial was to assess the effect of enamel sandblasting in addition to the acid-etch technique in reducing first-time failures of fixed mandibular retainers placed over a 12-month period. MATERIALS AND METHODS: Ethical approval was obtained. Participants were recruited in a single private practice. The primary outcome of this study was any first-time failure of the mandibular fixed retainer assessed at three timepoints over a 12-month period. Three consecutive teeth either on the left or right side (from lower canine-lower central incisor) were randomly allocated to the intervention (sandblasting) and the control (non-sandblasted). Randomization was performed using a centralized randomization service. The patients were randomized in blocks of four and eight with allocation concealment secured by contacting the sequence generator for group assignment. Blinding of either the patient or clinicians was not possible at time of placement of the mandibular retainer. RESULTS: One hundred and ninety-seven participants were randomized to receive enamel sandblasting (intervention) and non-sandblasting (control) in the region of the six anterior mandibular teeth in a split-mouth fashion. Participants were recruited between December 2018 to October 2020. The data for all participants were analysed resulting in 394 observations. Overall, the risk of first-time failure was 11.4%. No difference in first-time failures was observed between the intervention (sandblasting) and control (non-sandblasting) groups [hazard ratio (HR), 1.05; 95% confidence interval (CI), 0.59, 1.88, P = 0.88]. Males had a higher instant probability of first-time failures (HR, 3.18; 95% CI, 1.65-6.14; P < 0.01). Participants with a fair level of co-operation had a lower instant probability of first-time failures (HR, 0.37; 95% CI, 0.16-0.86; P = 0.02). There were no harms reported to either the participant or their dentition. CONCLUSIONS: No difference in the first-time failures of mandibular bonded retainers placed with conventional etch-bond technique with or without enamel sandblasting was observed. The overall risk of first-time failure was 11.4%. REGISTRATION: This trial was not registered prior to trial commencement.

Accurate gingival recession quantification using 3D digital dental models.

OBJECTIVES: To develop and validate a method for accurate quantitative assessment of gingival recessions based on superimposition of serial 3D digital models. MATERIALS AND METHODS: Gingival recessions of mild (0.5-2 mm) and increased (3-7 mm) severity were simulated on stone casts and surface models were created. The outlines of the gingival margins of the mild (A) and severe recessions (B) were compared to the original gingival margins following 3D best fit superimposition through a gold standard technique (GS), which used intact adjacent structures, and the tested method (CC), which used single tooth crowns at the position of recessions, as superimposition reference. The primary outcome was the distance between the most apical point of each corresponding gingival margin along the respective tooth long axis. RESULTS: For mild recessions, the median difference of the test methods (CC_A) from the reference method (GS_A) was 0.008 mm (IQR: 0.093; range: - 0.143, 0.147). For severe recessions, the median difference of the test method (CC_B) from the reference method (GS_B) was 0.009 mm (IQR: 0.091; range: - 0.170, 0.198). The proposed method (CC) showed very high intra- and inter-operator reproducibility (median: 0.025 and 0.033 mm, respectively). CONCLUSIONS: The suggested method offers highly accurate monitoring of gingival margin changes and diagnosis of gingival recessions using 3D digital dental models. The method is applicable irrespective of changes in tooth position or form, allowing for assessments over any time span. CLINICAL RELEVANCE: The accurate detection and visualization of gingival margin changes in 3D will enhance diagnosis and patient-doctor communication.

Consistent downregulation of the cleft lip/palate-associated genes IRF6 and GRHL3 in carcinomas.

Interferon Regulatory Factor 6 (IRF6) and Grainyhead Like Transcription Factor 3 (GRHL3) are transcription factors that orchestrate gene regulatory networks required for the balance between keratinocyte differentiation and proliferation. Absence of either protein results in the lack of a normal stratified epidermis with keratinocytes failing to stop proliferating and to terminally differentiate. Numerous pathological variants within IRF6 and GRHL3 have been identified in orofacial cleft-affected individuals and expression of the two transcription factors has been found to be often dysregulated in cancers. However, whether orofacial cleft-associated IRF6 and GRHL3 variants in patients might also affect their cancer risk later in life, is not clear yet. The fact that the role of IRF6 and GRHL3 in cancer remains controversial makes this question even more challenging. Some studies identified IRF6 and GRHL3 as oncogenes, while others could attribute tumor suppressive functions to them. Trying to solve this apparent conundrum, we herein aimed to characterize IRF6 and GRHL3 function in various types of carcinomas. We screened multiple cancer and normal cell lines for their expression, and subsequently proceeded with functional assays in cancer cell lines. Our data uncovered consistent downregulation of IRF6 and GRHL3 in all types of carcinomas analyzed. Reduced levels of IRF6 and GRHL3 were found to be associated with several tumorigenic properties, such as enhanced cell proliferation, epithelial mesenchymal transition, migration and reduced differentiation capacity. Based on our findings, IRF6 and GRHL3 can be considered as tumor suppressor genes in various carcinomas, which makes them potential common etiological factors for cancer and CLP in a fraction of CLP-affected patients.

Effect of hydration on the anatomical form of human dry skulls.

In radiology research soft tissues are often simulated on bone specimens using liquid materials such as water, or gel-like materials, such as ballistic gel. This study aimed to test the effect of hydration on the anatomical form of dry craniofacial bone specimens. Sixteen human dry skulls and 16 mandibles were scanned with an industrial scanner in dry conditions and after water embedding. Ten skulls were also embedded for different time periods (5 or 15 min). The subsequent 3D surface models were best-fit superimposed and compared by calculating mean absolute distances between them at various measurement areas. There was a significant, primarily enlargement effect of hydration on the anatomical form of dry skeletal specimens as detected after water embedding for a short time period. The effect was smaller in dry skulls (median 0.20 mm, IQR 0.17 mm) and larger in mandibles (median 0.56 mm, IQR 0.57 mm). The effect of different water embedding times was negligible. Based on the present findings, we suggest to shortly hydrate the skeletal specimens prior to reference model acquisition so that they are comparable to hydrated specimens when liquid materials are used as soft-tissue simulants for various radiologic research purposes.

Discovery and characterization of heterogeneous and multipotent fibroblast populations isolated from excised cleft lip tissue.

BACKGROUND: Regularly discarded lip tissue obtained from corrective surgeries to close the cleft lip represents an easily accessible and rich source for the isolation of primary fibroblasts. Primary fibroblasts have been described to show compelling similarities to mesenchymal stem cells (MSCs). Hence, cleft lip and palate (CLP) lip-derived fibroblasts could be thought as an intriguing cell source for personalized regenerative therapies in CLP-affected patients. METHODS: Initially, we thoroughly characterized the fibroblastic nature of the lip-derived mesenchymal outgrowths by molecular and functional assays. Next, we compared their phenotype and genotype to that of bone marrow-mesenchymal stem cells (BM-MSCs) and of human lung-derived fibroblasts WI38, by assessing their morphology, surface marker expression, trilineage differentiation potential, colony-forming (CFU) capacity, and immunomodulation property. Finally, to better decipher the heterogeneity of our CLP cultures, we performed a single cell clonal analysis and tested expanded clones for surface marker expression, as well as osteogenic and CFU potential. RESULTS: We identified intriguingly similar phenotypic and genotypic properties between CLP lip fibroblasts and BM-MSCs, which makes them distinct from WI38. Furthermore, our own data in combination with the complex anatomy of the lip tissue indicated heterogeneity in our CLP cultures. Using a clonal analysis, we discovered single cell-derived clones with increased levels of the MSC markers CD106 and CD146 and clones with variabilities in their commitment to differentiate into bone-forming cells and in their potential to form single cell-derived colonies. However, we were not able to gain clones possessing superior MSC-like capacities when compared to the heterogeneous parental CLP population. Additionally, all clones could still generate contractile forces and retained robust levels of the fibroblast specific marker FSP1, which was not detectable in BM-MSCs. CONCLUSIONS: Our results suggest that we isolate heterogeneous populations of fibroblasts from discarded CLP lip tissue, which show a prominently multipotent character in their entirety avoiding the need for elaborate subpopulation selections in vitro. These findings suggest that CLP lip fibroblasts might be a novel potential cell source for personalized regenerative medicine of clinical benefit for CLP patients.

Precision of a Hand-Held 3D Surface Scanner in Dry and Wet Skeletal Surfaces: An Ex Vivo Study.

Three-dimensional surface scans of skeletal structures have various clinical and research applications in medicine, anthropology, and other relevant fields. The aim of this study was to test the precision of a widely used hand-held surface scanner and the associated software's 3D model generation-error in both dry and wet skeletal surfaces. Ten human dry skulls and ten mandibles (dry and wet conditions) were scanned twice with an industrial scanner (Artec Space Spider) by one operator. Following a best-fit superimposition of corresponding surface model pairs, the mean absolute distance (MAD) between them was calculated on ten anatomical regions on the skulls and six on the mandibles. The software's 3D model generation process was repeated for the same scan of four dry skulls and four mandibles (wet and dry conditions), and the results were compared in a similar manner. The median scanner precision was 31 µm for the skulls and 25 µm for the mandibles in dry conditions, whereas in wet conditions it was slightly lower at 40 µm for the mandibles. The 3D model generation-error was negligible (range: 5-10 µm). The Artec Space Spider scanner exhibits very high precision in the scanning of dry and wet skeletal surfaces.

Gingival thickness threshold and probe visibility through soft tissue: a cross-sectional study.

OBJECTIVES: The aim was to retrieve the threshold of gingival thickness (GT), where the attribute of gingival translucency through probe visibility was altered. METHODS: In 200 patients, the soft tissue thickness was evaluated at both central mandibular incisors using ultrasound quantification (USD). Additionally, probe visibility was determined using a standard periodontal probe (PB) (CPU 15 UNC, Hu-Friedy), inserted 1 mm deep into the gingival sulcus. Frequencies and relative frequencies were calculated. Repeatability analyses and receiver operating characteristics (ROC) were conducted to determine the USD cut-off point for probe visibility. RESULTS: Regression model indicated that the probe was not visible at a thickness of 0.82 mm for the mandibular left central incisor (95% CIs 0.77, 0.86) and became visible at a thickness of 0.69 mm (95% CIs 0.65, 0.72). The respective values for the mandibular right central incisor were 0.82 mm (95% CIs 0.77, 0.87) and 0.70 mm (0.68, 0.74). ROC analysis confirmed the retrieved regression results by indicating the best fitting balance for specificity and sensitivity at a thickness of 0.8 mm for both mandibular incisors. CONCLUSIONS: In the frame of the current study, the data revealed that gingiva becomes non-transparent at a thickness of approximately 0.8 mm. CLINICAL RELEVANCE: Probe visibility at mandibular incisors for the discrimination between thin and thick soft tissues was correlated with a gingival thickness of 0.8 mm and a high repeatability.

Responses of the Mediterranean seagrass Cymodocea nodosa to combined temperature and salinity stress at the ionomic, transcriptomic, ultrastructural and photosynthetic levels.

The Little Neptune grass Cymodocea nodosa is a key seagrass species in the Mediterranean Sea, forming extensive and patchy meadows in shallow coastal and transitional ecosystems. In such habitats, high temperatures and salinities, separately and in combination, can be significant stressors in the context of climate change, particularly during heatwave events, and seawater desalination plant effluents. Despite well-documented negative, macroscopic effects, the underlying cellular and molecular processes of the combined effects of increasing temperature and salinities have remained largely elusive in C. nodosa - which are addressed by the present study. High salinity and high temperature, alone and in combination, affected ion equilibrium in the plant cells. Non-synonymous mutations marked the transcriptomic response to salinity and temperature stress at loci related to osmotic stress. Cell structure, especially the nucleus, chloroplasts, mitochondria and organization of the MT cytoskeleton, was also altered. Both temperature and salinity stress negatively affected photosynthetic activity as evidenced by ΔF/Fm', following an antagonistic interaction type. Overall, this study showed that all biological levels investigated were strongly affected by temperature and salinity stress, however, with the latter having more severe effects. The results have implications for the operation of desalination plants and for assessing the impacts of marine heat waves.

Effect of combined periodontal and orthodontic treatment of tilted molars and of teeth with intra-bony and furcation defects in stage-IV periodontitis patients: A systematic review.

AIM: To assess the effect of combined periodontal and orthodontic treatment (OT) in stage-IV periodontitis patients. MATERIALS AND METHODS: Three focused questions were addressed using the Population, Intervention, Comparison, Outcome, and Study Design criteria. Randomized controlled trials (RCTs), controlled clinical trials, follow-up studies, case series, and controlled/uncontrolled before/after studies were assessed for inclusion. Primary outcomes included mean changes in pocket probing depth (PPD) and clinical attachment level (CAL). Qualitative synthesis of results was performed and reported according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines. RESULTS: Out of 916 records, 1 retrospective case series study reported the effect of OT of tilted molars, 2 RCTs and 10 prospective and 2 retrospective case series studies reported the effect of OT of treated intra-bony defects and 0 articles reported the effect of OT of treated furcation defects. Mean PPD changes were reported in 14 articles, and mean CAL changes were reported in 8 articles. Risk of bias was high in both included RCTs, critical in nine articles, and serious in four articles. No articles included patient-reported outcomes, and three articles reported harms/adverse effects. CONCLUSIONS: Evidence is limited by (i) the lack or low number of included studies, (ii) the apparent methodological and clinical heterogeneity, and (iii) the high risk of bias of the retrieved studies. No solid conclusions could be drawn concerning OT in stage-IV periodontitis patients with respect to tilted molars, teeth with treated intra-bony defects, and teeth with treated furcation defects.

Bone turnover markers in gingival crevicular fluid and blood serum of patients with fixed orthodontic appliances.

AIM: Bone remodelling can be followed through the bone turnover markers (BTMs). Aim of the present study was to record the fluctuation of an osteoclastic and an osteoblastic BTM [C-terminal telopeptide of type I collagen (CTX) and N-terminal pro-peptide of type I pro-collagen (PINP), respectively] in both the gingival crevicular fluid (GCF) and the serum of orthodontic patients before and after the initial application of orthodontic forces. MATERIALS AND METHODS: Twenty-one Caucasian patients were prospectively evaluated. GCF and blood samples were collected in order to measure the selected biomarkers by ELISA at three time-points: exactly before, 5 days, and 14 days after bonding of the appliances. Standardized sample handling and patient preparation procedures were adopted in order to reduce pre-analytical variability. RESULTS: GCF and serum CTX levels were found to be independent of age, although higher in the serum of female subjects. PINP levels were found higher in the serum of patients ≥25 years old, as well as in the GCF of males. A positive correlation between serum and GCF baseline PINP levels was observed. LIMITATIONS: The effect of orthodontic treatment on bone remodelling might not be absolutely representative of the local bone microenvironment as the levels of the specific BTMs where measured within the GCF of the lower front teeth. CONCLUSIONS: This is the first time PINP and CTX have been evaluated in the GCF and serum of orthodontic patients with fixed appliances. No statistically significant alterations of CTX and PINP levels in the GCF and the serum of patients were recorded over time during the initial stages of orthodontic treatment.

Success and complication rate of miniscrew assisted non-surgical palatal expansion in adults - a consecutive study using a novel force-controlled polycyclic activation protocol.

INTRODUCTION: Bone-borne miniscrew assisted palatal expansion (MAPE) is a common technique to improve maxillary transverse deficiency in young adolescents. Adult patients usually present a challenge, as they often require additional surgical assisted maxillary expansion (SARPE). There is still no clear statement about non-surgical expansion in adult patients using this technique. The aim of this study was to evaluate the success and complication rate of non-surgical palatal expansion in adults utilizing MAPE with a novel force-controlled polycyclic expansion protocol (FCPC). METHODS: This consecutive study consisted of 33 adult patients with an average age of 29.1 ± 10.2 years (min. 18 years, max. 58 years), including one dropout patient. First, four miniscrews were inserted and after 12-weeks latency, the expander was placed and the FCPC protocol was applied (MAPE group). In case of missing expansion, a SARPE was performed (SARPE group). After maximum expansion, a cone beam CT was made and widening of the midpalatal suture was measured. The outcome variables were successful non-surgical expansion and, with sample size power above 80%, the odds of failed non-surgical expansion and associated complications were evaluated. The primary predictor variable was age. Statistical analysis was performed using R (Version 3.1) to calculate power, to construct various models for measuring the odds of requiring surgical intervention/complications, and others. RESULTS: Successful non-surgical expansion was achieved in 27 patients (84.4%), ranging from 18 to 49 years. Mean age differed significantly between both groups (26.8 ± 8.2 years vs. 41.3 ± 9.9 years; p < 0.001). Mean expansion at the anterior and posterior palate for the MAPE group was 5.4 ± 1.5 mm and 2.5 ± 1.1 mm, respectively. Among these subjects' complications were observed in 18.5%. Age significantly increased the odds of complications (p = 0.019). CONCLUSIONS: 1. The success rate of MAPE among individuals aged 18 to 49 years was 84.4%. 2. A V-shaped expansion pattern in the antero-posterior dimension was mostly observed. 3. Complications were significantly associated with age. 4. A careful expansion protocol seems to be beneficial to prevent unfavorable results in adult patients. TRIAL REGISTRATION: Consecutive cohort study, Review Board No. EK-2-2014/0016.

Lack of IRF6 Disrupts Human Epithelial Homeostasis by Altering Colony Morphology, Migration Pattern, and Differentiation Potential of Keratinocytes.

Variants within the gene encoding for the transcription factor Interferon Regulatory Factor 6 (IRF6) are associated with syndromic and non-syndromic Cleft Lip/Palate (CLP) cases. IRF6 plays a vital role in the regulation of the proliferation/differentiation balance in keratinocytes and is involved in wound healing and migration. Since a fraction of CLP patients undergoing corrective cleft surgery experience wound healing complications, IRF6 represents an interesting candidate gene linking the two processes. However, Irf6 function has been mainly studied in mice and knowledge on IRF6 in human cells remains sparse. Here, we aimed to elucidate the role of IRF6 in human postnatal skin- and oral mucosa-derived keratinocytes. To do so, we applied CRISPR/Cas9 to ablate IRF6 in two TERT-immortalized keratinocyte cultures, which we used as model cell lines. We show that IRF6 controls the appearance of single cells and colonies, with the latter being less cohesive in its absence. Consequently, IRF6 knockout keratinocytes often moved as single cells instead of a collective epithelial sheet migration but maintained their epithelial character. Lack of IRF6 triggered severe keratinocyte differentiation defects, which were already apparent in the stratum spinosum and extended to the stratum corneum in 3D organotypic skin cultures, while it did not alter their growth rate. Finally, proteomics revealed that most of the differentially expressed proteins in the absence of IRF6 could be associated with differentiation, cell-cell adhesion as well as immune response. Our data expand the knowledge on IRF6 in human postnatal keratinocytes, which will help to better understand IRF6-related pathologies.