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Background This study aimed to evaluate possible cytotoxic effects to gingival epithelial cells exposed to children toothpastes containing different detergent. Methods Tissues required for the isolation of human gingival epithelial cells were obtained by biopsy during the extraction of the impacted third molar tooth. Toothpaste solutions of different concentrations were prepared from five different children’s toothpastes with different detergent contents. Isolated gingival epithelial cells were stimulated with experimental groups consisting of toothpaste solutions (Colgate, Sensodyne, Splat, Nenedent, Perlodent) at different concentrations and a control group consisting of complete Dulbecco’s modified eagle medium. After the experiments, cell viability was evaluated using flow cytometry. 2 Way ANOVA was used to see the interaction effect of the main effects of toothpaste solution and concentration factors. Pairwise comparisons were made by Tukey post hoc tests. In the study, the significance level was taken as 0.05. Results As a result of the analysis, it was seen that the toothpaste solution and concentration factors and the interactions of these 2 factors were effective on the viable, early apoptotic, late apoptotic and necrotic cell rates. The statistically highest live cell ratios were detected in Splat’s toothpaste solutions (90.14% at 0.4% concentration) after the control group (90.82%) and the group with the lowest viability values was determined in Colgate group (75.74% at 0.4% concentration) (p Conclusions According to the results of the study, it was observed that toothpastes containing SLS affected the viability of cells more negatively than toothpastes with other detergent contents.

Amelogenesis imperfecta (AI) is a group of rare genetic diseases affecting the tooth enamel. AI is characterized by an inadequate quantity and/or quality of tooth enamel and can be divided into three major categories: hypoplastic, hypocalcified and hypomaturation types. Even though there are some overlapping phenotypes, hypomaturation AI enamel typically has a yellow to brown discoloration with a dull appearance but a normal thickness indicating a less mineralized enamel matrix. In this study, we recruited four Turkish families with hypomaturation AI and performed mutational analysis using whole exome sequencing. These analyses revealed two novel homozygous mutations in the KLK4 gene: a nonsense mutation in exon 3 (NM_004917.4:c.170C>A, p.(Ser57*)) was found in families 1, 2 and 3 and a missense mutation in exon 6 (c.637T>C, p.(Cys213Arg)) in family 4. Functional analysis showed that the missense mutation transcript could not translate the mutant protein efficiently or generated an unstable protein that lacked functional activity. The two novel inactivating KLK4 mutations we identified caused a hypomaturation AI phenotype similar to those caused by the four previously described KLK4 nonsense and frameshift mutations. This study improves our understanding of the normal and pathologic mechanisms of enamel formation.

Human ACP4 (OMIM*606362) encodes a transmembrane protein that belongs to histidine acid phosphatase (ACP) family. Recessive mutations in ACP4 cause non-syndromic hypoplastic amelogenesis imperfecta (AI1J, OMIM#617297). While ACP activity has long been detected in developing teeth, its functions during tooth development and the pathogenesis of ACP4-associated AI remain largely unknown. Here, we characterized 2 AI1J families and identified a novel ACP4 disease-causing mutation: c.774_775del, p.Gly260Aspfs*29. To investigate the role of ACP4 during amelogenesis, we generated and characterized Acp4R110C mice that carry the p.(Arg110Cys) loss-of-function mutation. Mouse Acp4 expression was the strongest at secretory stage ameloblasts, and the protein localized primarily at Tomes’ processes. While Acp4 heterozygous (Acp4+/R110C) mice showed no phenotypes, incisors and molars of homozygous (Acp4R110C/R110C) mice exhibited a thin layer of aplastic enamel with numerous ectopic mineralized nodules. Acp4R110C/R110C ameloblasts appeared normal initially but underwent pathology at mid-way of secretory stage. Ultrastructurally, sporadic enamel ribbons grew on mineralized dentin but failed to elongate, and aberrant needle-like crystals formed instead. Globs of organic matrix accumulated by the distal membranes of defective Tomes’ processes. These results demonstrated a critical role for ACP4 in appositional growth of dental enamel probably by processing and regulating enamel matrix proteins around mineralization front apparatus.

Mutations in Dentin Sialophosphoprotein (DSPP) are known to cause, in order of increasing severity, dentin dysplasia type-II (DD-II), dentinogenesis imperfecta type-II (DGI-II), and dentinogenesis imperfecta type-III (DGI-III). DSPP mutations fall into two groups: a 5′-group that affects protein targeting and a 3′-group that shifts translation into the −1 reading frame. Using whole-exome sequence (WES) analyses and Single Molecule Real-Time (SMRT) sequencing, we identified disease-causing DSPP mutations in 12 families. Three of the mutations are novel: c.53T>C/p.(Val18Ala); c.3461delG/p.(Ser1154Metfs*160); and c.3700delA/p.(Ser1234Alafs*80). We propose genetic analysis start with WES analysis of proband DNA to identify mutations in COL1A1 and COL1A2 causing dominant forms of osteogenesis imperfecta, 5′-DSPP mutations, and 3′-DSPP frameshifts near the margins of the DSPP repeat region, and SMRT sequencing when the disease-causing mutation is not identified. After reviewing the literature and incorporating new information showing distinct differences in the cell pathology observed between knockin mice with 5′-Dspp or 3′-Dspp mutations, we propose a modified Shields Classification based upon the causative mutation rather than phenotypic severity such that patients identified with 5′-DSPP defects be diagnosed as DGI-III, while those with 3′-DSPP defects be diagnosed as DGI-II.

Hereditary dentin defects are conventionally classified into three types of dentinogenesis imperfecta (DGI) and two types of dentin dysplasia (DD). Mutations in the dentin sialophosphoprotein (DSPP) gene have been identified to cause DGI type II and III and DD type II; therefore, these are not three different conditions, but rather allelic disorders. In this study, we recruited three families with varying clinical phenotypes from DGI-III to DD-II and performed mutational analysis by candidate gene analysis or whole-exome sequencing. Three novel mutations including a silent mutation (NM_014208.3: c.52-2del, c.135+1G>C, and c.135G>A; p.(Gln45=)) were identified, all of which affected pre-mRNA splicing. Comparison of the splicing assay results revealed that the expression level of the DSPP exon 3 deletion transcript correlated with the severity of the dentin defects. This study did not only expand the mutational spectrum of DSPP gene, but also advanced our understanding of the molecular pathogenesis impacting the severity of hereditary dentin defects.

Amelogenesis imperfecta (AI) is a group of rare genetic diseases affecting the tooth enamel. AI is characterized by an inadequate quantity and/or quality of tooth enamel and can be divided into three major categories: hypoplastic, hypocalcified and hypomaturation types. Even though there are some overlapping phenotypes, hypomaturation AI enamel typically has a yellow to brown discoloration with a dull appearance but a normal thickness indicating a less mineralized enamel matrix. In this study, we recruited four Turkish families with hypomaturation AI and performed mutational analysis using whole exome sequencing. These analyses revealed two novel homozygous mutations in the

Amelogenesis imperfecta (AI) is a rare genetic condition affecting the quantity and/or quality of tooth enamel. Hypomaturation AI is characterized by brownish-yellow discoloration with increased opacity and poorly mineralized enamel prone to fracture and attrition. We recruited three families affected by hypomaturation AI and performed whole exome sequencing with selected individuals in each family. Bioinformatic analysis and Sanger sequencing identified and confirmed mutations and segregation in the families. Family 1 had a novel homozygous frameshift mutation in GPR68 gene (NM_003485.3:c.78_83delinsC, p.(Val27Cysfs*146)). Family 2 had a novel homozygous nonsense mutation in SLC24A4 gene (NM_153646.4:c.613C>T, NP_705932.2:p.(Arg205*)). Family 3 also had a homozygous missense mutation in SLC24A4 gene which was reported previously (c.437C>T, p.(Ala146Val)). This report not only expands the mutational spectrum of the AI-causing genes but also improves our understanding of normal and pathologic amelogenesis.

Background: This study aimed to evaluate possible cytotoxic effects to gingival epithelial cells exposed to children toothpastes containing different detergent. Methods: Tissues required fort he isolation of human gingival epithelial cells were obtained by biopsy during the extraction of the impacted third molar tooth. Toothpaste solutions of different concentrations were prepared from five different children’s toothpastes with different detergent contents. Isolated gingival epithelial cells were stimulated with experimental groups consisting of toothpaste solutions (Colgate, Sensodyne, Splat, Nenedent, Perlodent) at different concentrations and a control group consissting of complete Dulbocco’s modified eagle medium. After the experiments, cell viability was evaluated using flow cytometry. Data analysis were done using One Way ANOVA test and Tukey post-hoc test. Results: In all experimental groups, there was a decrease in live cell rates and an increase in dead cell rates due to increased concentration. The statistically highest live cell ratios were detected in Splat’s toothpaste solutions after the control group and the group with the lowest viability values was determined in Colgate group (pConclusions: According to the results of the study, it was observed that toothpastes containing SLS affected the viability of cells more negatively than toothpastes with other detergent contents.

Amelogenesis imperfecta (AI) is an innate disorder that affects the formation and mineralization of the tooth enamel. When diagnosed with AI, one’s teeth can be hypoplastic (thin enamel), hypomature (normal enamel thickness but discolored and softer than normal enamel), hypocalcified (normal enamel thickness but extremely weak), or mixed conditions of the above. Numerous studies have revealed the genes that are involved in causing AI. Recently, ACP4 (acid phosphatase 4) was newly found as a gene causing hypoplastic AI, and it was suggested that mutant forms of ACP4 might affect access to the catalytic core or the ability to form a homodimer. In this study, a Korean and a Turkish family with hypoplastic AI were recruited, and their exome sequences were analyzed. Biallelic mutations were revealed in ACP4: paternal (NM_033068: c.419C>T, p.(Pro140Leu)) and maternal (c.262C>A, p.(Arg88Ser)) mutations in family 1 and a paternal (c.713C>T, p.(Ser238Leu)) mutation and de novo (c.350A>G, p.(Gln117Arg)) mutation in the maternal allele in family 2. Mutations were analyzed by cloning, mutagenesis, immunofluorescence, immunoprecipitation, and acid phosphatase activity test. Comparison between the wild-type and mutant ACP4s showed a decreased amount of protein expression from the mutant forms, a decreased ability to form a homodimer, and a decreased acid phosphatase activity level. We believe that these findings will not only expand the mutational spectrum of ACP4 but also increase our understanding of the mechanism of ACP4 function during normal and pathologic amelogenesis.

The goal of the present work was to revisit published data to test if genetic variation in interferon regulatory factor 6 (IRF6) is associated with children born with cleft lip with or without cleft palate (CL/P) for cases with positive history of maternal smoking. From the 573 individuals originally studied, this reanalysis focused on 57 who had a positive history of maternal smoking during pregnancy (39 born with CL/P and 18 born without CL/P). Seven IRF6 markers (rs4844880, rs2235371, rs2013162, ra861019, rs2073487, rs642961, and rs658860) were tested for over-transmission of alleles and an alpha of 0.05 was considered statistically significant. All individuals born with CL/P were homozygous for the wild type allele of rs2235371 in comparison to just two individuals born without clefts (p = 0.0000001). For rs861019, individuals born with CL/P were more likely to have the variant allele (p = 0.006). A similar trend was seen for rs642961 (p = 0.09). The results suggest that statistical evidence of over-representation of IRF6 alleles in individuals born with CL/P may be unveiled only when maternal smoking during pregnancy is used as the inclusion criterion in the analysis.

Background Detergents are the most commonly used compounds in toothpastes due to their foaming and cleaning peoperties. This study aimed to investigate the effects of children’s toothpastes with different detergent content on the viability, the osteogenic and chondrogenic differentiation potentials of human mesenchymal stem cells. Methods The necessary tissues for human periodontal ligament mesenchymal stem cells (hPDLMSCs) and human gingival mesenchymal stem cells (hGMSCs) isolation were obtained during extraction of 10 impacted third molar teeth. The viability of the cells stimulated with different concentratiaons of Colgate, Sensodyne, Splat, Nenedent, Perlodent toothpaste solutions and complete Dulbocco’s modified eagle medium (control group) were evaluated by using the flow cytometer. In addition, the osteogenic and chondrogenic differentiation potential of human gingival and periodontal ligament mesenchymal stem cells exposed to toothpaste solutions were examined morphologically. Datas were analyzed with IBM SPSS V23. One way ANOVA test was used to determine the differences between the groups for multiple comparisons, while the Tukey post-hoc test was used for pair wise comparisons in determining which groups differed. Results A higher percentage of cell viability was detected in Control group at 20 %, 50 % and 80 % (p = 0.000) on hGMSCs. After the Control group, the highest cell viability ratios were observed in the detergent-free Splat group (p = 0.000) followed by the Sensodyne experimental group containing CABP (p = 0.000). While the cell viability rates in Nenedent group was found significantly higher than the Perlodent group at other concentrations except for 20 % concentration (p = 0.000). Colgate group had the lowest percentage of cell viability among the experimental groups at all concentrations on hPDMSCs (p = 0.000). The highest live cell ratios was detected in Control group (p = 0.000), followed by Splat and Sensodyne groups (p = 0.000). The cell viability ratios at 50 % concentration were higher in Perlodent group than Nenedent group (p = 0.000). The highest osteogenic and chondrogenic differentiation potential of mesenchymal stem cells stimulated with different toothpaste was determined in Control and Splat group. Conclusions As a result of the findings, it was observed that toothpaste containing SLS had a more negative effect on the viability of the cells and the differentiation potentials than the other groups.

Background: The aim of this study was to assess the effect of traumatic dental injuries and dental caries on the Oral Health-Related Quality of Life (OHRQoL) of preschool children.Methods: The 13 questions and six domains of Early Childhood Oral Health Impact Scale (ECOHIS) were administered to 206 preschool children’s parents to measure Oral Health-Related Quality of Life. Type of traumatic dental injuries, dental caries (dmft scores), and anterior malocclusions traits of the children were recorded after the oral examination.Results: The children’s mean age was 4.09±0.97. Dental caries showed a negative impact on the overall Oral Health-Related Quality of Life mean and symptoms, function, psychological, parental distress, and family function domains. The presence of anterior malocclusion traits showed a negative effect only on the social interaction domain. Complicated traumatic dental injuries showed a negative effect on symptoms domain than uncomplicated traumatic dental injuries.Conclusion: Having a tooth with traumatic dental injuries and malocclusions have a negative impact on the Oral Health-Related Quality of Life of preschool children only one domain (child domain), although dental caries, has a strong association (child domain+family domain) with the Oral Health-Related Quality of Life of preschool children and their parents., Amaç: Bu çalışmanın amacı, travmatik diş yaralanmalarının (TDY) ve diş çürüğünün okul öncesi çocukların Ağız Sağlığı ile İlişkili Yaşam Kalitesi (OHRQoL) üzerindeki etkisini değerlendirmektir.Gereç ve yöntemler: Ağız Sağlığı ile İlişkili Yaşam Kalitesini ölçmek için 206 okul öncesi çocuğun velisine 13 soruluk Erken Çocukluk Çağı Ağız Sağlığı Etki Ölçeği (ECOHIS) uygulanmıştır. Çocukların travmatik diş yaralanmaları, diş çürüğü (dmft skorları) ve ön bölge maloklüzyon özelliklerine bakılmıştır.Bulgular: Çocukların yaş ortalaması 4.09 ± 0.97 bulunmuştur. Diş çürüğü, Ağız Sağlığı ile İlişkili Yaşam Kalitesi üzerinde olumsuz bir etki göstermiştir. Diş çürüğü; Ağız Sağlığı ile İlişkili Yaşam Kalitesi ortalaması, semptomları, fonksiyonu, psikolojisi, ailesel sıkıntı ve aile fonksiyon alanları üzerinde olumsuz bir etki göstermiştir. Ön bölgede maloklüzyon varlığı sadece sosyal etkileşim alanı üzerinde olumsuz bir etki göstermiştir. Komplike travmatik diş yaralanmaları, komplike olmayan travmatik diş yaralanmalarına göre semptomlar açısından daha olumsuz bir etki göstermiştir.Sonuç: Travmatik dental yaralanma ve maloklüzyon gözlenen dişin varlığı, okul öncesi çocukların Ağız Sağlığı ile İlişkili Yaşam Kalitesi üzerinde yalnızca çocuk alanında olumsuz bir etkiye sahiptir; ancak, diş çürüğü, hem çocuk alanında hem aile alanında Ağız Sağlığı ile İlişkili Yaşam Kalitesi ile güçlü bir ilişkiye sahiptir. Anahtar Kelimeler: Ağız Sağlığı ile İlişkili Yaşam Kalitesi, dental travma, diş çürüğü, Erken Çocukluk Çağı Ağız Sağlığı Etki Ölçeği, okul öncesi çocuklar

Dental enamel malformations, or amelogenesis imperfecta (AI), can be isolated or syndromic. To improve the prospects of making a successful diagnosis by genetic testing, it is important that the full range of genes and mutations that cause AI be determined. Defects in WDR72 (WD repeat-containing protein 72; OMIM *613214) cause AI, type IIA3 (OMIM #613211), which follows an autosomal recessive pattern of inheritance. The defective enamel is normal in thickness, severely hypomineralized, orange-brown stained, and susceptible to attrition. We identified 6 families with biallelic WDR72 mutations by whole exome sequence analyses that perfectly segregated with the enamel phenotype. The novel mutations included 3 stop-gains [NM_182758.2: c.377G>A/p.(Trp126*), c.1801C>T/p.(Arg601*), c.2350A>T/p.(Arg784*)], a missense mutation [c.1265G>T/p.(Gly422Val)], and a 62,138–base pair deletion (NG_017034.2: g.35441_97578del62138) that removed WDR72 coding exons 3 through 13. A previously reported WDR72 frameshift was also observed [c.1467_1468delAT/p.(Val491Aspfs*8)]. Three of the affected patients showed decreased serum pH, consistent with a diagnosis of renal tubular acidosis. Percentiles of stature and body weight varied among 8 affected individuals but did not show a consistent trend. These studies support that WDR72 mutations cause a syndromic form of AI and improve our ability to diagnose AI caused by WDR72 defects.

OBJECTIVE: Amelogenesis imperfecta (AI) is a rare hereditary disorder affecting the quality and quantity of the tooth enamel. The purpose of this study was to identify the genetic etiology of hypoplastic AI families based on the candidate gene approach. Materials and methods We recruited three Turkish families with hypoplastic AI and performed a candidate gene screening based on the characteristic clinical feature to find the pathogenic genetic etiology. RESULTS: The candidate gene sequencing of the LAMB3 gene for family 1 revealed a heterozygous nonsense mutation in the last exon [c.3431C > A, p.(Ser1144*)]. FAM20A gene sequencing for families 2 and 3 identified a homozygous deletion [c.34_35delCT, p.(Leu12Alafs*67)] and a homozygous deletion-insertion (c.1109 + 3_1109 + 7delinsTGGTC) mutation, respectively. CONCLUSION: The candidate gene approach can be successfully used to identify the genetic etiology of the AI in some cases with characteristic clinical features. CLINICAL RELEVANCE: Identification of the genetic etiology of the AI will help both the family members and dentist understand the nature of the disorder. Characteristic clinical feature can suggest possible genetic causes.

Made available in DSpace on 2018-12-11T16:52:03Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-04-01 Objective: The goal of the present work was to use dental conditions that have been independently associated with cleft lip and palate (CL/P) as a tool to identify a broader collection of individuals to be used for gene identification that lead to clefts. Study design: We studied 1573 DNA samples combining individuals that were born with CL/P or had tooth agenesis, supernumerary teeth, molar incisor hypomineralization, or dental caries with the goal to identify genetic associations. We tested 2 single-nucleotide polymorphisms that were located in the vicinity of regions suggested to contribute to supernumerary teeth. Overrepresentation of alleles were determined for combinations of individuals as well as for each individual phenotypic group with an α of.05. Results: We determined that the allele C of rs622260 was overrepresented in all individuals studied compared with a group of unrelated individuals who did not present any of the conditions described earlier. When subgroups were tested, associations were found for individuals with hypomineralization. Conclusions: Although we did not test this hypothesis directly in the present study, based on associations reported previously, we believe that CL/P is actually a syndrome of alterations of the dentition, and considering it that way may allow for the identification of genotype-phenotype correlations that may be useful for clinical care. Department of Pedodontics School of Dentistry Istanbul University Department of Pedodontics School of Dentistry Istanbul Medipol University Department of Oral Biology University of Pittsburgh School of Dental Medicine Department of Pediatric Dentistry UNESP Department of Pediatric Dentistry University of Pittsburgh School of Dental Medicine Department of Pediatric Dentistry UNESP

Background/Aims: Enamel-renal syndrome is characterized by nephrocalcinosis, enamel defects, gingival hyperplasia and eruption failures. It has been recently identified that recessive mutations in the FAM20A gene result in amelogenesis imperfecta (AI)-gingival fibromatosis. The aim of this research to determine whether AI patients with known ­FAM20A mutations also have nephrocalcinosis. Methods: Complete oral and radiological examinations were performed for all participating family members. Renal examinations were performed using ultrasound. Results: The teeth were evaluated for severe loss, and multiple eruption failures were evident from the clinical and radiological examinations. Unexpected extensive and fast crown resorption was found by radiological examination. Renal ultrasound revealed bilateral nephrocalcinosis in both affected individuals. Recessive FAM20A mutations can cause nephrocalcinosis in addition to the oral phenotype. Conclusion: AI patients with similar clinical phenotypes and FAM20A mutations should be examined for nephropathy even if they lack pertinent symptoms. Nephrology referral is warranted for patients who have clinical phenotypes related to AI-gingival fibromatosis even if they are not symptomatic.

Amaç: Bu çalışmada diş çürüğü deneyimi ile ergen bireyler ve anneler arasındaki tutarlılık duygusu (SOC) ilişkisinin saptanması ve tutarlılık duygusunun bireylerin oral hijyenleri üzerindeki etkisinin incelenmesi amaçlanmıştır. Gereç ve Yöntemler: Bu çalışma 11-14 yaş grubu arasındaki sağlıklı 100 ergen birey ve anneleri dahil edilerek oral klinik muayeneleri DMFT indeksi ve Silness - Löe plak indeksi kullanılarak yapılmıştır. Demografik verilerden kardeş sayısı, anne eğitim durumu, baba eğitim durumu ve aile geliri değerlendirilmiştir. Tutarlılık duygusu, tutarlılık duygu skalasının 13 sorudan oluşan Türkçe’ye uyarlanmış kısa anket versiyonu kullanılarak uygulanmıştır. Yanıtlar, her soru için 7’li Likert ölçeği ile değerlendirilmiştir. Tutarlılık duygusu ile DMFT, plak indeksi, demografik veriler arasındaki ilişki istatistiksel olarak Spearman Korelasyon Analizi kullanılarak incelenmiştir. Bulgular: Çocukların SOC değeriyle annelerin SOC değeri arasında orta düzeyde anlamlı ilişki bulunmuştur. Kardeş sayısı ve aile geliriyle çocukların SOC ve annelerin SOC değerleri arasında istatistiksel olarak anlamlı ilişki yoktur. Anne eğitim durumlarıyla çocukların SOC değeri arasında zayıf düzeyde anlamlı ilişki bulunurken, anne eğitim durumlarıyla annelerin SOC değeri arasında anlamlı ilişki bulunmamıştır. Çocukların DMFT değerleriyle çocukların SOC ve annelerin SOC değerleri arasında anlamlı bir ilişki bulunmazken çocukların DMFT değerleriyle çocuk plak indeksi arasında ise zayıf düzeyde anlamlı bir ilişki mevcuttur.Sonuç: Sonuç olarak anneyle ergen bireylerin tutarlılık duyguları arasında ilişki gözlenirken, diş çürüğü ve tutarlılık duygusu arasında ilişki bulunamamıştır., Aim: The aim of this study was to investigate relationship between caries experience and sense of coherence (SOC) among adolescent and their mothers and to investigate effects of SOC on oral hygiene of individuals and their experience of caries.Material and Methods: In this study, 100 healthy adolescents aged 11-14 years and their mothers were evaluated and oral clinical examinations were performed using DMFT index and Silness - Lee plaque index. The number of siblings, parents’ educational level, and family income were recorded as a demographic data. Sense of coherence was implemented using a short questionnaire version adapted to Turkish, consisting of 13 questions of the coherence emotion scale. The answers were evaluated with a 7-point Likert scale for each question. The correlation between SOC and DMFT, plaque index and demographic data were evaluated statistically using Spearman Correlation Analysis (p

Amelogenesis imperfecta (AI) is a collection of genetic disorders affecting the quality and/or quantity of tooth enamel. More than 20 genes are, so far, known to be responsible for this condition. In this study, we recruited 3 Turkish families with hypomaturation AI. Whole-exome sequence analyses identified disease-causing mutations in each proband, and these mutations cosegregated with the AI phenotype in all recruited members of each family. The AI-causing mutations in family 1 were a novel AMELX mutation [NM_182680.1:c.143T>C, p.(Leu48Ser)] in the proband and a novel homozygous MMP20 mutation [NM_004771.3:c.616G>A, p.(Asp206Asn)] in the mother of the proband. Previously reported compound heterozygous MMP20 mutations [NM_004771.3:c.103A>C, p.(Arg35=) and c.389C>T, p.(Thr130Ile)] caused the AI in family 2 and family 3. Minigene splicing analyses revealed that the AMELX missense mutation increased exonic definition of exon 4 and the MMP20 synonymous mutation decreased exonic definition of exon 1. These mutations would trigger an alteration of exon usage during RNA splicing, causing the enamel malformations. These results broaden our understanding of molecular genetic pathology of tooth enamel formation.

Background/purpose: Dental pulp stem cells (DPSCs) play a crucial role in the tis-sue healing process through odontoblast like cell differentiation. The aim of this study was to evaluate the biocompatibility and compare the potential invitro cytotoxic effects of NeoMTA Plus, ProRootMTA and Biodentine on human dental pulp stem cells (hDPSCs). Materials and methods: To assess the effects of NeoMTA Plus, ProRoot MTA and Biodentine ex-tracts at 1st, 3rd and 7th d on hDPCs, cell populations was determined by flow cytometry using an Annexin V detection kit. The data were analyzed statistically using the Kruskal-Wallis test. A p < 0.05 was considered as statistically significant. Results: All groups showed cell viability similar to that of the control group on 1st, 3rd and 7th d. Although Biodentine exhibited higher cell viability rates than the other material groups, no statistically significant differences were noted between the sampled days (p > 0.05). Conclusion: All materials extracts are not cytotoxic and do not induce apoptosis in the hDPSCs. These results suggest that all the tested materials can lead to positive outcomes when used as reparative biomaterials. (c) 2020 Association for Dental Sciences of the Republic of China. Publishing services by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons. org/licenses/by-nc-nd/4.0/).

Purpose The objective of this case series was to determine the oral, dental and craniofacial features of patients with EvC syndrome. Material and methods Eight patients with EvC syndrome were enrolled. A complete family history, pedigree analysis, detailed medical history were collected. Findings of clinical examination, including craniofacial and orodental manifestations, and radiological investigations were thoroughly studied. Results All eight patients had characteristic face, hypertrophic frenulum, conical and peg-shaped teeth, hypodontia of deciduous and/or permanent teeth and also skeletal dysplasia, small chest, short stature and hypoplastic nails. Additionally dysmorphic filtrum, serrated appearance of gingiva, diastema, enamel hypoplasia, microdontia, taurodontism, single rooted permanent molar, delayed eruption and high caries rate were observed with varying degrees. Cephalometric evaluation revealed skeletal Class III growth pattern in four subjects and Class II growth pattern in one subject. Conclusion Evaluation of craniofacial and orodental anomalies of EvC syndrome is required for accurate differential diagnosis from other congenital syndromes.

Aim: Although there are many materials that can be used for retrograde filling in surgical endodontics, none of them can be regarded as an ideal material yet. The purpose of this study was to compare the surface topography of three different root-end filling materials. Methods: 36 extracted single rooted human incisor teeth were cleaned and decoronated to standardized 10 mm root lengths. The root segments were prepared and 2 mm apical resection were performed. The samples were randomly separeted to three groups (Group A: Ca(OH)2, Group B: MTA Angelus, Group C: ProRoot MTA), each one of them was comprised of 12 roots. Materials were placed as 2 mm apical barriers and obturated with guttapercha and AH-Plus sealer. Each group was dimidiated into two subgroups (A1,A2,B1,B2,C1,C2). Groups A1,B1,C1 were stored in normal saline (NS), groups A2,B2,C2 were stored in neutral phosphate buffer saline (NPBS) solution and samples were incubated at 370C for 2 weeks. Stereomicroscope (32X) was used to photograph the root-end filling. Results: All specimens demonstrated white crystals formation and sediment over the root-end filling materials and on the superficial border of the root-end cavities’ wall as a white plaque. A2,B2,C2 samples had more crystal sediment on root-end fillings than samples A1,B1,C1. Dissolution and corrosion were observed in groups A1, A2. Conclusions: The results of this study revealed that calcium hydroxide is more resorbable than MTA Angelus and ProRoot MTA. The crystals formation and precipitation were observed in neutral phosphate buffer saline solution which were more than normal saline solution for all groups as a hydroxiapatite crystals.

WOS: 000458956100004 PubMed ID: 30506946 Amelogenesis imperfecta (AI) is a collection of isolated (non-syndromic) inherited diseases affecting dental enamel formation or a clinical phenotype in syndromic conditions. We characterized three consanguineous AI families with generalized irregular hypoplastic enamel with rapid attrition that perfectly segregated with homozygous defects in a novel gene: RELT that is a member of the tumor necrosis factor receptor superfamily (TNFRSF). RNAscope in situ hybridization of wild-type mouse molars and incisors showed specific Relt mRNA expression by secretory stage ameloblasts and by odontoblasts. Relt(-/-) mice generated by CRISPR/Cas9 exhibited incisor and molar enamel malformations. Relt(-/-) enamel had a rough surface and underwent rapid attrition. Normally unmineralized spaces in the deep enamel near the dentino-enamel junction (DEJ) were as highly mineralized as the adjacent enamel, which likely altered the mechanical properties of the DEJ. Phylogenetic analyses showed the existence of selective pressure on RELT gene outside of tooth development, indicating that the human condition may be syndromic, which possibly explains the history of small stature and severe childhood infections in two of the probands. Knowing a TNFRSF member is critical during the secretory stage of enamel formation advances our understanding of amelogenesis and improves our ability to diagnose human conditions featuring enamel malformations. National Institute of Dental and Craniofacial Research [DE015846DE016276]; National Research Foundation of Korea [NRF-2017R1A2A2A05069281NRF-2018R1A5A2024418] National Institute of Dental and Craniofacial Research, Grant/Award Number: DE015846DE016276; National Research Foundation of Korea, Grant/Award Number: NRF-2017R1A2A2A05069281NRF-2018R1A5A2024418

Aplasia of lacrimal and salivary glands (ALSG) is a rare autosomal dominant inherited disease, characterized by aplasia, atresia, or hypoplasia of the lacrimal and salivary systems with variable expressivity. The purpose of this study was to identify genetic etiology of an ALSG family. We recruited a Turkish family with ALSG and performed a mutational analysis, based on the candidate gene approach, to clarify the molecular genetic etiology. The candidate gene sequencing of the FGF10 gene identified a novel heterozygous nonsense mutation (c.237G > A, p.Trp79*) in the exon 1. The identified novel mutation would result in a haploinsufficiency of the FGF10, because of nonsense-mediated mRNA decay caused by a premature stop codon. This report further confirms that ALSG is caused by the haploinsufficiency of functional FGF10. Identification of the genetic etiology of the ALSG will help both the family members and dentist understand the nature of the disorder. Therefore, it will positively motivate oral health care to avoid further destruction of the tooth due to the lack of salivary production.

Truncation mutations in FAM83H (family with sequence similarity 83, member H) cause autosomal dominant hypocalcified amelogenesis imperfecta (ADHCAI), but little is known about FAM83H function and the pathogenesis of ADHCAI. We recruited three ADHCAI families and identified two novel (p.Gln457*; p.Lys639*) and one previously documented (p.Q452*) disease-causing FAM83H mutations. We generated and characterized Fam83h-knockout/lacZ-knockin mice. Surprisingly, enamel thickness, density, Knoop hardness, morphology, and prism patterns were similar in Fam83h (+/+), Fam83h (+/-), and Fam83h (-/-) mice. The histology of ameloblasts in all stages of development, in both molars and incisors, was virtually identical in all three genotypes and showed no signs of pathology, although the Fam83h (-/-) mice usually died after 2 weeks and rarely survived to 7 weeks. LacZ expression in the knockin mice was used to report Fam83h expression in the epithelial tissues of many organs, notably in skin and hair follicles, which manifested a disease phenotype. Pull-down studies determined that FAM83H dimerizes through its N-terminal phospholipase D-like (PLD-like) domain and identified potential FAM83H interacting proteins. Casein kinase 1 (CK1) interacts with the FAM83H PLD-like domain via an F(270)-X-X-X-F(274)-X-X-X-F(278) motif. CK1 can phosphorylate FAM83H in vitro, and many phosphorylation sites were identified in the FAM83H C-terminus. Truncation of FAM83H alters its subcellular localization and that of CK1. Our results support the conclusion that FAM83H is not necessary for proper dental enamel formation in mice, but may act as a scaffold protein that localizes CK1. ADHCAI is likely caused by gain-of-function effects mediated by truncated FAM83H, which potentially mislocalizes CK1 as part of its pathological mechanism.

Nonsyndromic dentin defects classified as type II dentin dysplasia and types II and III dentinogenesis imperfecta are caused by mutations in DSPP (dentin sialophosphoprotein). Most reported disease‐causing DSPP mutations occur within the repetitive DPP (dentin phosphoprotein) coding sequence. We characterized the DPP sequences of five probands with inherited dentin defects using single molecule real‐time (SMRT) DNA sequencing. Eight of the 10 sequences matched previously reported DPP length haplotypes and two were novel. Alignment with known DPP sequences showed 32 indels arranged in 36 different patterns. Sixteen of the 32 indels were not represented in more than one haplotype. The 25 haplotypes with confirmed indels were aligned to generate a tree that describes how the length variations might have evolved. Some indels were independently generated in multiple lines. A previously reported disease‐causing DSPP mutation in Family 1 was confirmed and its position clarified (c.3135delC; p.Ser1045Argfs*269). A novel frameshift mutation (c.3504_3508dup; p.Asp1170Alafs*146) caused the dentin defects in Family 2. A COL1A2 (c.2027G>A or p.Gly676Asp) missense mutation, discovered by whole‐exome sequencing, caused the dentin defects in Family 3. We conclude that SMRT sequencing characterizes the DPP repeat region without cloning and can improve our understanding of normal and pathological length variations in DSPP alleles.

Objective: The aim of this in vitro study was to evaluate antimicrobial activities of three different pulp capping materials; Biodentine, mineral trioxide aggregate (MTA) Angelus, and Dycal against Enterococcus faecalis and their durability with time. Materials and Methods: Direct contact test was used for the assessment. Three sets of sealers were mixed and placed on microtiter plate wells: One set was used within 20 min of recommended setting time while others were used after 24-h and 1-week. E. faecalis suspension was placed directly on the materials for 1 h and then transferred to another plate with fresh media. Nine wells of bacteria without the tested cements served as the positive control. One well of the tested cements without bacteria served as the negative control. Bacterial growth was evaluated by a temperature-controlled microplate spectrophotometer for 1-h intervals among 24 h. Data were analyzed using Kruskal–Wallis test. Results: All tested materials showed less bacterial density than the control group. MTA, Biodentine, and Dycal showed significantly higher bacterial density than the control group in freshly mixed samples (P < 0.05). And MTA showed significantly higher antibacterial activity than Dycal (P < 0.05). In 24 h, materials did not show any differences (P > 0.05). MTA and Biodentine samples showed significant differences than Dycal; MTA also showed higher antibacterial activity than control in 1-week samples (P < 0.05). Conclusion: While freshly mixed MTA showed the best antibacterial activity over time, Biodentine had shown similar antibacterial activity to MTA.

Biyoteknoloji alanindaki gelismeler, medikal arastirmalarda gen tedavisini on plana cikarmis ve gunumuz genetik prensiplerinin agiz dis sagligini gelistirmek icin kullanilmasina yol acmistir. Bu yeni teknolojilerinin dis hekimligi ile bulusmasini; restoratif biyometaryallerin uretiminde kaydedilen bilimsel asamalar, doku muhendisligi, molekuler biyolojinin kullanimi ve insan genom projesi saglamistir. Gen tedavisi, defektli genin ozel bir yontem kullanilarak tedavi edici bir gen ile degistirilmesi veya tamir edilmesidir. Bircok hastaligin tedavisinde ve genetik bozukluklarin duzeltilmesinde kullanilabilmektedir. Gen tedavisinin dis hekimliginde dentofasiyal uygulama alanlari arasinda kemik rejenerasyonu, DNA bazli immunizasyon (curuk ve periodontal hastaliklar), agri tedavisi, otoimmun hastaliklarin tedavisi ve kanser tedavisi yer almaktadir. Genel olarak arastirmalarda yeni vektor sistemlerinin ve tasima metotlarinin gelistirilmesi, bunlarin test edilmesi, vektor transduksiyonunu iceren mekanizmalarin acikliga kavusturulmasi ve en iyi sonuc verebilecek yaklasimlarin klinik deneylerde kullanilmasina calisilmaktadir. Gectigimiz 15 yilda gen ve ilac tasiyici sistemler icin cesitli vektorler gelistirilmeye calisilmistir. Gen tedavisi, dis hekimligi alaninda bilimsel bazda onemli etkiye sahiptir. Bu derlemede gen tedavisi metotlari, ilgili vektorler, gen tedavisinin dis hekimliginde klinik etkisi ve konuya iliskin bilimsel bilgilerin guncellenmesi amaclanmistir. Anahtar Kelimeler: dis hekimligi, genetik, gen tedavisi, genetik vektorler GENE THERAPY IN DENTISTRY (REVIEW) ABSTRACT Advances in biotechnology have brought gene therapy to a very important and active area of medical research and c urrently genetic principles are being applied along to improve oral health. Scientific advances in restorative biomaterials, tissue engineering, molecular biology and the human genome project provide the basis for the introduction of new technologies into dentistry. Gene therapy is a particular method by which missing or defective genes are replaced or repaired by normal genes. It can be used to treat wide range of diseases and correct genetics disorders. In dentistry the application of gene therapy to dentofacial problems includes bone regeneration, DNA based immunization (for caries and periodontal disease), pain, for treating auto immune disease and cancer. The general approach is to develop new vector systems and delivery methods, test them, uncover the mechanisms involved in vector transduction, and use the most promising approaches in clinical trials. For the past 15 years various vectors have been designed for improved gene and drug delivery. This field is having significant impact on topics related to dental practice. The aim of this review is to provide an update on gene therapy methods, its clinical implication in dentistry and scientific information associated with gene therapy. Key Words: dentistry; genetics; gene therapy; genetic vectors

Purpose The aim of this study is to evaluate clinical success rate of resin-based fissure sealants applied at Istanbul University, Department of Pediatric Dentistry after 1 year of application. Materials and Methods Children with at least one pair of caries-free permanent first molars with deep pits and fissures were included in the study. The ages of children ranged from 7-13 (mean age: 9.2±1.22). Resin-based fissure sealant was applied to the 322 fissures of the first permanent molars in 100 children. 12 months after the application, children were recalled for examination. Recall examinations were carried out by the same dentist. Fissure sealants were evaluated at 12th month by using Ryge criteria: Retention (R), precence of caries (PC), marginal adaptation (MA), marginal discoloration (MD) and cracking (C). Results 12 months after the application, all of the children were recalled and fissure sealants were examinated. Our findigs are: R: Alpha 95 (29.5%), Bravo: 143 (44.4%), Charlie: 84 (26%); PC: Alpha 274, Bravo 48 (14.9%); MD: Alpha 322; MA: Alpha 279, Bravo 43 (13.3%); C: Alpha 321, Bravo 1 (0.3%). Conclusion The preventive effects of the sealant are only maintained as long as it remains completely intact and bonded in place. After applying fissure sealants, patients must be recalled and sealants must be checked to provide retention and marginal adaptation.

In this case report of autosomal recessive pigmented hypomaturation amelogenesis imperfecta (AI), we identify a novel homozygous missense mutation (g.165151 T>G; c.1317 T>G; p.Leu436 Arg) in SLC24A4, a gene encoding a potassium-dependent sodium-calcium exchanger that is critical for hardening dental enamel during tooth development.

Objective Hereditary defects in tooth enamel formation, amelogenesis imperfecta (AI), can be non-syndromic or syndromic phenotype. Integrins are signaling proteins that mediate cell–cell and cell–extracellular matrix communication, and their involvement in tooth development is well known. The purposes of this study were to identify genetic cause of an AI family and molecular pathogenesis underlying defective enamel formation. Materials and Methods We recruited a Turkish family with isolated AI and performed mutational analyses to clarify the underlying molecular genetic etiology. Results Autozygosity mapping and exome sequencing identified a novel homozygous ITGB6 transversion mutation in exon 4 (c.517G>C, p.Gly173Arg). The glycine at this position in the middle of the βI-domain is conserved among a wide range of vertebrate orthologs and human paralogs. Clinically, the enamel was generally thin and pitted with pigmentation. Thicker enamel was noted at the cervical area of the molars. Conclusions In this study, we identified a novel homozygous ITGB6 mutation causing isolated AI, and this advances the understanding of normal and pathologic enamel development.

The aim of this paper was to review the literature and present a case of regional odontodysplasia with details of clinical and radiographic features,thereby facilitating management of this rare anomaly. A 6-year-old girl was referred to our department with chief complaint of missing or unerupted permanent maxillary left central, lateral and canine teeth. At the affected area, the gingiva was enlarged, fibrous, and tense. Radiographic examination shows abnormally formed teeth with short roots. After 3 year follow up with temporary prosthetic rehabilitation, periodontal surgery was applied.Following forced eruption and levelling, abnormal tooth eruption and root development were observed in these three teeth. After root canal treatment with ProRoot MTA, an intracanal fiber post and the access cavity was restored with composite resin and zirconia ceramic crown. The choice of the best treatment option for ROD depends on the time of diagnosis, present/existing symptoms,functional and esthetic needs,and available treatment modalities.

Amaç: Travmatik dental yaralanmalar, dünyadaki çocuk popülasyonunda sıklıkla görülen problemlerden birisidir. Bu çalışmanın amacı 6-12 yaş arası okul çağı çocuklarında görülen travmatik dental yaralanmaların meydana gelme sıklığı ve travma ile ilişkili faktörlerin değerlendirilmesidir. Gereç ve Yöntemler:Bu çalışma, Nevşehir ve İstanbul illerinde bulunan toplamda beş farklı okuldan, ortalama yaşı 8.9±0.84 olan 897 çocuk (472 kız , 425 erkek) üzerinde yapılmıştır. Kayıtlar diş hekimleri tarafından cinsiyet, yaş, travmanın tipi, etkilenen dişin tipi, sağlanan tedavi, travmanın nedeni ve dental bilgilere göre (diş eksikliği, supernumerer diş, baş başa kapanış, overjet, openbite, deepbite, croosbite) değerlendirildi ve ailedeki ebeveynlerin çalışma durumu ile kardeş sayısının sosyoekonomik ve sosyokültürel olarak verileri kaydedildi. İstatistiksel değerlendirmeler p

Enamel-renal syndrome is characterized by nephrocalcinosis, enamel defects, gingival hyperplasia and eruption failures. It has been recently identified that recessive mutations in the FAM20A gene result in amelogenesis imperfecta (AI)-gingival fibromatosis. The aim of this research to determine whether AI patients with known -FAM20A mutations also have nephrocalcinosis.Complete oral and radiological examinations were performed for all participating family members. Renal examinations were performed using ultrasound.The teeth were evaluated for severe loss, and multiple eruption failures were evident from the clinical and radiological examinations. Unexpected extensive and fast crown resorption was found by radiological examination. Renal ultrasound revealed bilateral nephrocalcinosis in both affected individuals. Recessive FAM20A mutations can cause nephrocalcinosis in addition to the oral phenotype.AI patients with similar clinical phenotypes and FAM20A mutations should be examined for nephropathy even if they lack pertinent symptoms. Nephrology referral is warranted for patients who have clinical phenotypes related to AI-gingival fibromatosis even if they are not symptomatic.

Ectodermal dysplasia (ED) is a hereditary disease characterized by anomalies in the structures of ectodermal origin. This report aims to determine the oral features of patients with ED syndrome and to present treatment approaches. Because of nutrition, speech and aesthetical problems due to the lack of teeth 22 patients with ED syndrome referred to faculty dentistry clinic. Facial physiognomy was typical for ED and the facial height was decreased due to the vertical dimension and the delalet exfoliation of primary teeth. Also anodontia, hypodontia, delayed eruption, diffuse enamel hypoplasia, conical tooth structure, talon cusps, microdontia, and, transposition in teeth were seen in patients. In most cases, prosthesis were applied to improve the facial aesthetics, speech and oral function due to the common hypodontia. The primary goals of the dental treatment of patients with ED are enhancing aesthetics and improving masticatory function. The oral rehabilitation of the patients’ needs multidisciplinary dental treatment.

Background Periodontal ligament (PDL) healing and long term prognosis of replanted avulsed teeth should rely on several factors including length of extra-oral dry time and type of the storage medium. The status of periodontal ligament is critical for the healing of replanted teeth. Different substances have been used for root surface treatment to promote formation of PDL and increase the survival of avulsed teeth submitted to replantation. Aim The purpose of this study was to assess the effect of recombinant basic fibroblast growth factor 2 (bFGF) and enamel matrix derivative (EMD) on root resorption after delayed replantation. Design 18 freshly extracted single-rooted incisor and premolar teeth were extracted from the beagle dogs and immersed in whole bovine milk for 45 and 60 min (n = 3 each). Following storage period, sockets washed and teeth were treated with bFGF and EMD and replanted into the sockets. After 8 weeks, dogs were sacrificed, specimens processed to 4-μm thick serial sections for histopathologic examination and morphometric assessments. Thus, the proportions of the roots that exhibited signs of surface resorption, inflammatory resorption, and replacement resorption, that is, ankylosis and normal PDL were noted. Results The percentage of root resorption was in the following order: EMD>milk>bFGF for 45 min and milk>EMD>bFGF for 60 min. For all groups, teeth stored 60 min showed significantly higher incidence of PDL resorption than those stored for 45 min (P

Mutations in the WNT10A gene were first detected in the rare syndrome odonto-onycho-dermal dysplasia (OODD, OMIM257980) but have now also been found to cause about 35–50% of selective tooth agenesis (STHAG4, OMIM150400), a common disorder that mostly affects the permanent dentition. In our random sample of tooth agenesis patients, 40% had at least one mutation in the WNT10A gene. The WNT10A Phe228Ile variant alone reached an allele frequency of 0.21 in the tooth agenesis cohort, about 10 times higher than the allele frequency reported in large SNP databases for Caucasian populations. Patients with bi-allelic WNT10A mutations have severe tooth agenesis while heterozygous individuals are either unaffected or have a mild phenotype. Mutations in the coding areas of the WNT10B gene, which is co-expressed with WNT10A during odontogenesis, and the WNT6 gene which is located at the same chromosomal locus as WNT10A in humans, do not contribute to the tooth agenesis phenotype. © 2014 Wiley Periodicals, Inc.

Aim Some of the commonly encountered dental emergencies are traumatic dental injuries. The aim of this study was to evaluate traumatic dental injuries with regard to age, gender, etiology, classification of trauma, teeth involved, place of injury, and treatment received by patients who were referred over a 2-year period to the Istanbul University, Faculty of Dentistry, Clinics of Pedodontics, Istanbul, Turkey. Materials and Methods The study was based on the clinical data of 154 patients (337 traumatized teeth). The children were examined clinically for dental injuries by the same investigator. The following information was recorded: age, gender, etiology, localization, place, number of injured teeth, type of trauma, type of tooth, time elapsed between injury and treatment, and treatment provided. Traumatized teeth were assessed according to the classification of the World Health Organization slightly modified. Results A total of 154 patients aged 1–13 years presented a total of 337 traumatized teeth (255 permanent and 82 primary). 94 boys (61%) and 60 girls (39%) with a mean age of 7.91 ± 3.15 years participated in the study. Dental injuries were frequent in the 6–12 year age group. The most common type of dental injuries recorded was luxation injuries (43.3%), uncomplicated crown fractures (20.5%), and complicated crown fractures (19.4%). The main causes were falls (55.2%) and being struck by an object (22.1%). The most frequent treatment was examination only (31.8%). Conclusions In childhood, a correct diagnosis may affect the management methods and prevent the occurrence of complications after treatment for traumatic dental injuries. Therefore, continuing education programs offering the latest updates in the management of traumatized teeth should be provided for dental and medical practitioners.

The aim of this clinical case series is to present a diagnosis and different treatment methods of patients in different ages with amelogenesis imperfecta (AI) as well as further treatments during a 3-6 years follow-up period. A number of 31 patients (16 female, 15 male with a mean age of 10.77 ± 2.65 years) with AI have been examined for the study group between 2007 and 2010 years. A detailed anamnesis was recorded, followed by a clinical and radiological assessment of oral health. The types of AI classified for each patient according to clinical and radiographic evaluation. The main complaints of patients, presence of dental caries and dental anomalies were noted. Necessary treatments had been planned for the individual cases of AI. A number of 19 patients had hypoplastic (HP) form, and 10 patients showed hypomaturation (HM) form of AI, while one patient showed hypocalcified form of AI and one patient had HM-HP form with taurodontism. Main complaints were chiefly related to dissatisfactory esthetics and dental sensitivity. Caries prevalence index was 93.5%. Mean decayed, missing, filling permanent teeth (DMF) and DMF surface (DMFS) were found as 2.74 ± 1.71 and 6.23 ± 3.99; df (decayed, filling primary teeth) and dfs (decayed, filling primary teeth surface) were found as 3.12 ± 2.85 and 5.24 ± 4.97, respectively. All patients received individual clinical care, including preventive, restorative, and prosthetic treatments. Patients have scheduled for regular follow-up in every 3 months. Composite restorations were used as the most common treatment (25 patients, 80.6%). The treatment plan should be based on patient's age, type of defects and individual needs of the patients. Necessary treatment plan is essential, not only due to functional and aesthetic reasons, but also for the positive psychological impact on young patients.

Evidence suggests caries experience is higher in children with asthma. This study compared caries experience in asthmatic and non-asthmatic children and defined whether variation in the distribution of caries experience differed between the two groups and was dependent on the presence of genetic variation in enamel formation genes. Children with asthma were recruited at the Istanbul University, Faculty of Medicine, Department of Paediatrics, Division of Paediatric Allergy and Pulmonary Diseases, and non-affected children were recruited at the Istanbul University, Faculty of Dentistry, Department of Paedodontics. Cases (N = 100) were defined as children between the ages of 6 and 12 years with asthma and controls (N = 100) as children without asthma. Cases and controls were matched by sex and age. All study subjects received a complete dental exam, provided demographic and other caries and asthma risk factors data, and a saliva sample for DNA extraction. Caries experience was defined based on DMFT/dmft and DMFS/dmfs scores. Genotypes of 11 SNPs were selected in intronic regions of enamel development genes. PCR with TaqMan chemistry was used for genotyping all selected markers. Association between caries experience (caries-free versus caries affected) depending on asthma status and SNPs was tested with PLINK by logistic regression, adjusting by risk, and other preventive measures. p values below 0.0045 (0.05/11) were considered statistically significant. Logistic regression analysis showed an association between AMBN rs4694075 and caries experience (p = 2.525e−007). This study provides, for the first time, evidence that ameloblastin is associated with caries in asthmatic children.

Amelogenesis imperfecta (AI) can be either isolated or part of a larger syndrome. Junctional epidermolysis bullosa (JEB) is a collection of autosomal-recessive disorders featuring AI associated with skin fragility and other symptoms. JEB is a recessive syndrome usually caused by mutations in both alleles of COL17A1, LAMA3, LAMB3, or LAMC2. In rare cases, heterozygous carriers in JEB kindreds display enamel malformations in the absence of skin fragility (isolated AI). We recruited two kindreds with autosomal-dominant amelogenesis imperfecta (ADAI) characterized by generalized severe enamel hypoplasia with deep linear grooves and pits. Whole-exome sequencing of both probands identified novel heterozygous mutations in the last exon of LAMB3 that likely truncated the protein. The mutations perfectly segregated with the enamel defects in both families. In Family 1, an 8-bp deletion (c.3446_3453del GACTGGAG) shifted the reading frame (p.Gly 1149Glufs*8). In Family 2, a single nucleotide substitution (c.C3431A) generated an in-frame translation termination codon (p.Ser1144*). We conclude that enamel formation is particularly sensitive to defects in hemidesmosome/basement-membrane complexes and that syndromic and non-syndromic forms of AI can be etiologically related.

Amelogenesis imperfecta is a group of rare inherited disorders that affect tooth enamel formation, quantitatively and/or qualitatively. The aim of this study was to identify the genetic etiologies of two families presenting with hypomaturation amelogenesis imperfecta. DNA was isolated from peripheral blood samples obtained from participating family members. Whole exome sequencing was performed using DNA samples from the two probands. Sequencing data was aligned to the NCBI human reference genome (NCBI build 37.2, hg19) and sequence variations were annotated with the dbSNP build 138. Mutations in MMP20 were identified in both probands. A homozygous missense mutation (c.678T>A; p.His226Gln) was identified in the consanguineous Family 1. Compound heterozygous MMP20 mutations (c.540T>A, p.Tyr180* and c.389C>T, p.Thr130Ile) were identified in the non-consanguineous Family 2. Affected persons in Family 1 showed hypomaturation AI with dark brown discoloration, which is similar to the clinical phenotype in a previous report with the same mutation. However, the dentition of the Family 2 proband exhibited slight yellowish discoloration with reduced transparency. Functional analysis showed that the p.Thr130Ile mutant protein had reduced activity of MMP20, while there was no functional MMP20 in the Family 1 proband. These results expand the mutational spectrum of the MMP20 and broaden our understanding of genotype-phenotype correlations in amelogenesis imperfecta.

We have previously shown that AQP5 and BTF3 genetic variation and expression in whole saliva are associated with caries experience suggesting that these genes may have a functional role in protecting against caries. To further explore these results, we tested ex vivo if variants in these genes are associated with subclinical dental enamel mineral loss. DNA and enamel samples were obtained from 53 individuals. Enamel samples were analyzed for Knoop hardness of sound enamel, integrated mineral loss after subclinical carious lesion creation, and change in integrated mineral loss after remineralization. DNA samples were genotyped for single nucleotide polymorphisms using TaqMan chemistry. Chi-square and Fisher's exact tests were used to compare individuals above and below the mean sound enamel microhardness of the cohort with alpha of 0.05. The A allele of BTF3 rs6862039 appears to be associated with harder enamel at baseline (p = 0.09), enamel more resistant to demineralization (p = 0.01), and enamel that more efficiently regain mineral and remineralize (p = 0.04). Similarly, the G allele of AQP5 marker rs3759129 and A allele of AQP5 marker rs296763 are associated with enamel more resistant to demineralization (p = 0.03 and 0.05, respectively). AQP5 and BTF3 genetic variations influence the initial subclinical stages of caries lesion formation in the subsurface of enamel. United States Department of Health & Human Services National Institutes of Health (NIH) - USA

Non-syndromic amelogenesis imperfecta (AI) is a collection of isolated inherited enamel malformations that follow X-linked, autosomal-dominant, or autosomal-recessive patterns of inheritance. The AI phenotype is also found in syndromes. We hypothesized that whole-exome sequencing of AI probands showing simplex or recessive patterns of inheritance would identify causative mutations among the known candidate genes for AI. DNA samples obtained from 12 unrelated probands with AI were analyzed. Disease-causing mutations were identified in three of the probands: a novel single-nucleotide deletion in both KLK4 alleles (g.6930delG; c.245delG; p.Gly82Alafs*87) that shifted the reading frame, a novel missense transition mutation in both MMP20 alleles (g.15390A>G; c.611A>G; p.His204Arg) that substituted arginine for an invariant histidine known to coordinate a structural zinc ion, and a previously described nonsense transition mutation in a single allele of FAM83H (c.1379G>A; g.5663G>A; p.W460*). Erupted molars and cross-sections from unerupted parts of the mandibular incisors of Mmp20 null mice were characterized by scanning electron microscopy. Their enamel malformations closely correlated with the enamel defects displayed by the proband with the MMP20 mutation. We conclude that whole-exome sequencing is an effective means of identifying disease-causing mutations in kindreds with AI, and this technique should prove clinically useful for this purpose.

Caries is a multifactorial disease and little is still known about the host genetic factors influencing susceptibility. Our previous genome-wide linkage scan has identified the interval 5q12.1–5q13.3 as linked to low caries susceptibility in Filipino families. Here we fine-mapped this region in order to identify genetic contributors to caries susceptibility. Four hundred and seventy-seven subjects from 72 pedigrees with similar cultural and behavioral habits and limited access to dental care living in the Philippines were studied. DMFT scores and genotype data of 75 single-nucleotide polymorphisms were evaluated in the Filipino families with the Family-Based Association Test. For replication purposes, a total 1,467 independent subjects from five different populations were analyzed in a case-control format. In the Filipino cohort, statistically significant and borderline associations were found between low caries experience and four genes spanning 13 million base pairs (PART1, ZSWIM6, CCNB1, and BTF3). We were able to replicate these results in some of the populations studied. We detected PART1 and BTF3 expression in whole saliva, and the expression of BTF3 was associated with caries experience. Our results suggest BTF3 may have a functional role in protecting against caries. Fil: Shimizu, T.. Nihon University of Dentistry; Japón Fil: Deeley, K.. University of Pittsburgh; Estados Unidos Fil: Briseño Ruiz, J.. University of Pittsburgh; Estados Unidos Fil: Faraco Junior, I. M.. University of Pittsburgh; Estados Unidos Fil: Poletta, Fernando Adrián. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. CEMIC-CONICET. Centro de Educaciones Médicas e Investigaciones Clínicas "Norberto Quirno". CEMIC-CONICET.; Argentina Fil: Brancher, J. A.. Pontifical Catholic University of Paraná; Brasil Fil: Pecharki, G. D.. Pontifical Catholic University of Paraná; Brasil Fil: Küchler, E. C.. Universidade Federal Fluminense; Brasil Fil: Tannure, P. N.. Universidade Federal do Rio de Janeiro; Brasil Fil: Lips, A.. Universidade Federal do Rio de Janeiro; Brasil Fil: Vieira, T. C. S.. Universidade Federal Fluminense; Brasil Fil: Patir, A.. Istanbul Medipol Universit; Turquía Fil: Yildirim, M.. Istanbul University; Turquía Fil: Mereb, J. C.. Centro de Educación Médica e Investigaciones Clínicas “Norberto Quirno”; Argentina Fil: Resick, J. M.. University of Pittsburgh; Estados Unidos Fil: Brandon, C. A.. University of Pittsburgh; Estados Unidos Fil: Cooper, M. E.. University of Pittsburgh; Estados Unidos Fil: Seymen, F.. Istanbul University; Turquía Fil: Costa, M. C.. Universidade Federal do Rio de Janeiro; Brasil Fil: Granjeiro, J. M.. Universidade Federal Fluminense; Brasil Fil: Trevilatto, P. C.. Pontifical Catholic University of Paraná; Brasil Fil: Orioli, I. M.. Universidade Federal do Rio de Janeiro; Brasil. Centro de Educación Médica e Investigaciones Clínicas “Norberto Quirno”; Argentina Fil: Castilla, Eduardo Enrique. Instituto Oswaldo Cruz; Brasil. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. CEMIC-CONICET. Centro de Educaciones Médicas e Investigaciones Clínicas "Norberto Quirno". CEMIC-CONICET.; Argentina Fil: Marazita, M. L.. University of Pittsburgh; Estados Unidos Fil: Vieira, A. R.. University of Pittsburgh; Estados Unidos

The runt-related transcription factor 2 gene (RUNX2), which is also known as CBFA1, is a master regulatory gene in bone formation. Mutations in RUNX2 have been identified in cleidocranial dysplasia (CCD) patients. CCD is a rare autosomal dominant skeletal dysplasia that is characterized by delayed closure of cranial sutures, aplastic or hypoplastic clavicle formation, short stature, and dental anomalies, including malocclusion, supernumerary teeth, and delayed eruption of permanent teeth. In this study, we recruited three de novo CCD families and performed mutational analysis of the RUNX2 gene as a candidate gene approach. The mutational study revealed three disease-causing mutations: a missense mutation (c.674G>A, p.Arg225Gln), a frameshift mutation (c.1119delC, p.Arg374Glyfs*), and a nonsense mutation (c.1171C>T, p.Arg391*). Clinical examination revealed a unique dental phenotype (no typical supernumerary teeth, but duplication of anterior teeth) in one patient. We believe that this finding will broaden the understanding of the mechanism of supernumerary teeth formation and CCD-related phenotypes.