The application of zebrafish model in the study of cleft lip and palate development: A systematic review.

Objective: Craniofacial growth and development are more than a scientific curiosity; it is of tremendous interest to clinicians. Insights into the genetic etiology of cleft lip and palate development are essential for improving diagnosis and treatment planning. The purpose of this systematic review was to utilize a zebrafish model to highlight the role of the IRF6 gene in cleft lip and palate development in humans. Data: This review adhered to the guidelines outlined in the PRISMA statement. Nine studies were included in the analysis. Sources: This study used major scientific databases such as MEDLINE, EMBASE, Web of Science, and the Zebrafish Information Network and yielded 1275 articles. Two reviewers performed the screening using COVIDENCE™ independently, and a third reviewer resolved any conflicts. Study selection: After applying the inclusion and exclusion criteria and screening, nine studies were included in the analysis. The Systematic Review Center for Laboratory Animal Experimentation's (SYRCLE's) risk-of-bias tool was used to assess the quality of the included studies. Results: The main outcome supports the role of the IRF6 gene in zebrafish periderm development and embryogenesis, and IRF6 variations result in cleft lip and palate development. The overall SYRCLE risk of bias was low-medium. Conclusion: In conclusion, this review indicated the critical role of the IRF6 gene and its downstream genes (GRHL3, KLF17, and ESRP1/2) in the development of cleft lip and palate in zebrafish models. Genetic mutation zebrafish models provide a high level of insights into zebrafish craniofacial development. Clinical relevance: this review provides a productive avenue for understanding the powerful and conserved zebrafish model for investigating the pathogenesis of human cleft lip and palate.

Accuracy of and dental students' preferences toward manual and digital cephalometric landmark identification: A randomized cross-over study.

OBJECTIVE: To evaluate dental students' perceptions of manual and digital cephalometric landmark identification methods based on their preferences, difficulty level, and procedure time required to provide insights into the future of dental education, considering incorporating digital technology in dental schools. MATERIALS AND METHODS: Fifty-five second-year dental students were randomly divided into two groups: (1) group A, students who performed manual landmark identification first, followed by digital method; and (2) group B, students who performed digital method first, followed by manual method. The duration of the procedure was recorded. Subsequently, all students completed a questionnaire regarding the difficulty they experienced using a visual analog scale and their preferences. Landmark identification accuracy was measured. RESULTS: Digital landmark identification was preferred by 93% of students. The mean procedure time for digital method was significantly lower than that of manual method (13.00 ± 5.60 vs. 9.70 ± 4.60; p = 0.002). Group B completed manual and digital methods in a shorter time than group A. Group A experienced less difficulty with manual procedure than group B. However, statistically significant differences were not observed in the difficulty level of digital technique. A statistically significant difference in the mean accuracy was shown in favor of the manual method. However, this difference is clinically insignificant (p = 0.001). CONCLUSIONS: Students considered digital method to be effective for learning and preferred it over manual method. Furthermore, digital landmark identification demonstrated better performance and was faster than manual method, suggesting that this must be incorporated in undergraduate dental education.

How Effective Are Non-Frictional Techniques Compared to Sliding Techniques in the Retraction of Upper Anterior Teeth When Using Buccal Fixed-Appliance Therapy? A Systematic Review.

Methods for retracting the anterior teeth are divided into frictional methods and non-frictional methods. However, evidence regarding the superiority of one technique over the other is still lacking in the available literature. Therefore, we aimed to evaluate the current evidence regarding the effectiveness of frictional methods of anterior teeth retraction compared to the non-frictional ones. The extracted data included the mechanism of application of the retraction force and its intensity, the observation period, follow-up records, and outcome measures. Ten studies were included in this review; the results did not favor a specific technique regarding the rate of orthodontic tooth movement and loss of anchorage during canine retraction, although a preference was shown for the sliding technique in the rate of en-masse retraction (0.74 versus 0.39 mm/month) and the anchorage control during the retraction of the incisors (0.5 versus 0.1 mm/month). The control of the incisor's torque during the en-masse retraction was higher when frictionless techniques were used (-12° versus -7°). Regarding the rate of orthodontic tooth movement, the non-frictional technique is characterized by a high sensitivity to the quality of the design, and the sliding technique was generally effective. As for controlling the torque of the incisors, the preference is for the non-frictional technique. Overall, there is a need to conduct more studies with an appropriate design.

Facial Soft Tissue Characteristics Among Sagittal and Vertical Skeletal Patterns: A Cone-Beam Computed Tomography Study.

Background Facial esthetics depend on the skeletal and dental structures underlying variable facial soft tissue thickness. In this social context, determining the relationship between external soft tissue and underlying skeletal and dental hard tissue is essential for detailed orthodontic diagnosis and treatment planning. Objective This study aims to measure facial soft tissue thickness in different sagittal and vertical skeletal patterns. Methodology This is an observational study utilizing pre-existing cone-beam computed tomography (CBCT) images of 170 subjects (110 females and 60 males) with a mean age group of 37.45 ± 13.83 years. CBCT images were then classified sagittally based on the point A-Nasion-point B (ANB) angle from Steiner's analysis into skeletal Class I, Class II, and Class III. Furthermore, vertical patterns were grouped based on the Frankfort-mandibular plane angle (FMA) from Tweed's analysis into hyperdivergent, hypodivergent, and normodivergent facial types. One-way ANOVA was used to compare the means of facial soft tissue thickness between the skeletal groups, followed by Tukey's post-hoc test for individual comparison. Multinomial logistic regression analysis was used to test the association between gender, age, and skeletal groups. The significance level was 0.05. Results One-way ANOVA revealed statistically significant differences in both sagittal and vertical groups (p≤0.05). Tukey's post hoc analysis showed that the skeletal Class III group has increased soft tissue thickness in the subnasale, upper lip, and mention compared to Class I and Class II subjects. Moreover, the hypodivergent group demonstrated increased soft tissue thickness in gnathion and mentioned landmarks in relation to the other groups. Multinomial logistic regression analysis showed significant differences between groups according to both gender and sagittal skeleton patterns (p≤0.05), with males less likely to be in Class II. Conclusions Skeletal Class III and hypodivergent groups have thicker soft tissue in specific facial landmarks. Sexual dimorphism was marked in soft tissue measurements.

Assessment of mandibular asymmetry in different skeletal malocclusions and vertical patterns in adult individuals: A cone-beam computed tomography study.

OBJECTIVES: The study aims to measure the mandibular condylar height (CH), ramus height (RH), total height (CH+RH), asymmetry index, and condylar volume (Cvol) in individuals with different anteroposterior and vertical skeletal discrepancies. MATERIALS AND METHODS: The study sample consisted of 131 subjects (60 females and 71 males) with a mean age of 35.06 ± 12.79 years. Pre-existing CBCT images were divided into groups according to the anteroposterior and vertical skeletal discrepancies. The investigator analyzed the data using t-tests to assess the mandibular bilateral sides of the individuals and gender differences. The mean difference between groups was determined using a one-way analysis of variance (ANOVA). The Chi-square test was used to study the association between the asymmetry index and groups. RESULTS: Each individual's bilateral sides exhibited statistically significant differences in CH, RH, and Cvol (P = 0.033, P = 0.039, P = 0.005, respectively), but not in CH+RH (P = 0.458). There were, however, statistically significant gender differences in CH+RH (P < 0.001). Skeletal Class III and hypodivergent groups revealed the highest linear and volumetric values compared to other groups. The asymmetry index was increased in CH (P = 0.006) and Cvol (P = 0.002) in skeletal Class II subjects. CONCLUSIONS: Significant differences in CH, RH, and Cvol were found on the right and left sides of the same individual. This study found increased linear and volumetric values in males, skeletal Class III, and hypodivergent subjects. Class II individuals had an increased CH and Cvol asymmetry index. This study highlights in-depth knowledge of mandibular asymmetry, which is extremely important to achieve an accurate diagnosis and provide the best treatment outcome.

Synergistic roles of Wnt modulators R-spondin2 and R-spondin3 in craniofacial morphogenesis and dental development.

Wnt signaling plays a critical role in craniofacial patterning, as well as tooth and bone development. Rspo2 and Rspo3 are key regulators of Wnt signaling. However, their coordinated function and relative requirement in craniofacial development and odontogensis are poorly understood. We showed that in zebrafish rspo2 and rspo3 are both expressed in osteoprogenitors in the embryonic craniofacial skeleton. This is in contrast to mouse development, where Rspo3 is expressed in osteoprogenitors while Rspo2 expression is not observed. In zebrafish, rspo2 and rspo3 are broadly expressed in the pulp, odontoblasts and epithelial crypts. However, in the developing molars of the mouse, Rspo3 is largely expressed in the dental follicle and alveolar mesenchyme while Rspo2 expression is restricted to the tooth germ. While Rspo3 ablation in the mouse is embryonic lethal, zebrafish rspo3-/- mutants are viable with modest decrease in Meckel's cartilage rostral length. However, compound disruption of rspo3 and rspo2 revealed synergistic roles of these genes in cartilage morphogenesis, fin development, and pharyngeal tooth development. Adult rspo3-/- zebrafish mutants exhibit a dysmorphic cranial skeleton and decreased average tooth number. This study highlights the differential functions of Rspo2 and Rspo3 in dentocranial morphogenesis in zebrafish and in mouse.

Impact of low-frequency coding variants on human facial shape.

The contribution of low-frequency variants to the genetic architecture of normal-range facial traits is unknown. We studied the influence of low-frequency coding variants (MAF < 1%) in 8091 genes on multi-dimensional facial shape phenotypes in a European cohort of 2329 healthy individuals. Using three-dimensional images, we partitioned the full face into 31 hierarchically arranged segments to model facial morphology at multiple levels, and generated multi-dimensional phenotypes representing the shape variation within each segment. We used MultiSKAT, a multivariate kernel regression approach to scan the exome for face-associated low-frequency variants in a gene-based manner. After accounting for multiple tests, seven genes (AR, CARS2, FTSJ1, HFE, LTB4R, TELO2, NECTIN1) were significantly associated with shape variation of the cheek, chin, nose and mouth areas. These genes displayed a wide range of phenotypic effects, with some impacting the full face and others affecting localized regions. The missense variant rs142863092 in NECTIN1 had a significant effect on chin morphology and was predicted bioinformatically to have a deleterious effect on protein function. Notably, NECTIN1 is an established craniofacial gene that underlies a human syndrome that includes a mandibular phenotype. We further showed that nectin1a mutations can affect zebrafish craniofacial development, with the size and shape of the mandibular cartilage altered in mutant animals. Findings from this study expanded our understanding of the genetic basis of normal-range facial shape by highlighting the role of low-frequency coding variants in several novel genes.

An Irf6-Esrp1/2 regulatory axis controls midface morphogenesis in vertebrates.

Irf6 and Esrp1 are important for palate development across vertebrates. In zebrafish, we found that irf6 regulates the expression of esrp1 We detailed overlapping Irf6 and Esrp1/2 expression in mouse orofacial epithelium. In zebrafish, irf6 and esrp1/2 share expression in periderm, frontonasal ectoderm and oral epithelium. Genetic disruption of irf6 and esrp1/2 in zebrafish resulted in cleft of the anterior neurocranium. The esrp1/2 mutant also developed cleft of the mouth opening. Lineage tracing of cranial neural crest cells revealed that the cleft resulted not from migration defect, but from impaired chondrogenesis. Analysis of aberrant cells within the cleft revealed expression of sox10, col1a1 and irf6, and these cells were adjacent to krt4+ and krt5+ cells. Breeding of mouse Irf6; Esrp1; Esrp2 compound mutants suggested genetic interaction, as the triple homozygote and the Irf6; Esrp1 double homozygote were not observed. Further, Irf6 heterozygosity reduced Esrp1/2 cleft severity. These studies highlight the complementary analysis of Irf6 and Esrp1/2 in mouse and zebrafish, and identify a unique aberrant cell population in zebrafish expressing sox10, col1a1 and irf6 Future work characterizing this cell population will yield additional insight into cleft pathogenesis.

Salivary alkaline phosphatase activity and chronological age as indicators for skeletal maturity.

OBJECTIVES: To investigate the relationship between salivary alkaline phosphatase activity (ALP), protein concentration, and chronological age with cervical vertebral maturation stages (CVMS) as noninvasive biomarkers for skeletal maturity assessment. MATERIALS AND METHODS: This cross-sectional study included 79 subjects (48 females, 31 males; 7 to 23 years old) categorized into five CVMS based on lateral cephalographs evaluated by three examiners. ALP activity and protein concentration in unstimulated whole saliva were compared among five CVMS. The association between age and CVMS was assessed and five multinomial logistic regression models were utilized to predict CVMS based on salivary ALP activity, protein concentration, and chronological age. RESULTS: Salivary ALP reached the peak at early pubertal stage and then declined with a significant difference between CVMS I and CVMS II (P < .001) and between CVMS I and CVMS V (P = .004). A significant positive correlation between age and CVMS was found (rs = 0.763, P < .001). The models' overall correct classification rates for predicting CVMS were 32.9% using protein concentration, 35.4% using ALP activity, and 53.2% using both ALP activity and age. CONCLUSIONS: The combination of salivary ALP activity and chronological age may provide the best CVMS prediction.

The effect of orthodontic therapy on periodontal health: a review of the literature.

Objectives. This review aims to evaluate the effect of orthodontic therapy on periodontal health. Data. Original articles that reported on the effect of orthodontic therapy on periodontal health were included. The reference lists of potentially relevant review articles were also sought. Sources. A literature search was conducted using the databases, Medline, EMBASE, Cochrane Library, Web of Science, Google Scholar, and Scopus databases for relevant studies. The search was carried out by using a combined text and the MeSH search strategies: using the key words in different combinations: "periodontal disease," "orthodontics" and "root resorption." This was supplemented by hand-searching in peer-reviewed journals and cross-referenced with the articles accessed. Articles published only in English language were included. Letters to the Editor, historical reviews and unpublished articles were not sought. Conclusions. Within the limitations of the present literature review, it was observed that there is a very close inter-relationship between the periodontal health and the outcome of orthodontic therapy.