Molar Tissue Research Articles

Effects of premature contact in maxillary alveolar bone in rats: relationship between experimental analyses and a micro scale FEA computational simulation study.

The aim of the investigation was to evaluate the maxillary alveolar bone morphology, bone architecture, and bone turnover in relation to the mechanical strain distribution in rats with dental premature contact. Fifty 2-month-old male Wistar rats were used. The premature contact group (N=40) received a unilateral (right side) resin cementation on the occlusal surface of the upper first molar. The animals were distributed in 4 subgroups according to the periods of euthanasia: 7, 14, 21, and 28 days after cementation (N=10, for each period). For the control group (N=10), the teeth were kept without resin, featuring a normal occlusion. The pieces including the upper first molars, alveolar bone, and periodontal tissue were processed to histological and immunohistochemical evaluation of RANK-L and TRAP protein expression. A three-dimensional bone microarchitecture analysis was performed, where the heads of animals were scanned using microtomography and analyzed using CT-Analyser software (Bruker, Kontich, Belgium). In the computer simulation by finite element analysis, two micro-scaled three-dimensional finite element models of first molar and dentoalveolar tissues were constructed, in representation of control and premature contact groups, using Materialise MIMICS Academic Research v18 (Materialise, Leuven, Belgium). The analysis was set to simulate a maxillary molar biting during the power stroke phase. The total deformation, equivalent strain, and minimum principal strain distribution were calculated. The expression of RANK-L and TRAP presented higher positive ratio in the 7-day period compared to the control group. The three-dimensional morphometry showed decrease of bone volume in the premature contact, with significant values between the control and the 7-day and 14-day groups (P = 0.007). In FEA, the premature contact model presented a uniform compressive strain distribution in the alveolar bone crest compared to a non-uniform compressive strain distribution in the control model. The results from FEA, 3D bone microarchitecture, and histological and immunohistochemical analyses showed that a model with dental traumatic occlusion resulted in changes of alveolar bone mechanobiology and, consequently, its morphology. These results could be applied in dental treatment planning bringing biological and mechanical feedback to provide an effective mechanism to obtain physiological bone loss responses. Furthermore, this association between experimental and computational analyses will be important to figure out the alveolar bone response to mechanical stimulation in different clinical conditions.

The Effect of Ethynil Estradiol and Desogestrel on Proliferation and Apoptosis Hydatidiform Mole Trophoblast Cell

Background: To explore the effect of ethynil estradiol and desogestrel on proliferation and apoptosis hydatidiform mole trophoblast cell.
 Methods: From April 2008 until March 2009, we collected 15 samples of hydatidiform mole tissue. Trophoblast cells was isolated and culture at RPMI 20% FBS medium. Only 7 samples (46%) shown good growth. Cell was identified using cytology and β HCG test. Trophoblast cells good quality than devided into three groups observation. First group get ethynil estradiol 10 nmol/mL, second group get desogestrel 100 nmol/mL, Third group get DMSO 1%. Cells incubated and observe at 24, 48, 72, 96 hours. Cell cycle, apoptosis and β HCG was evaluated at each time observation. Cell cycle evaluation using BD cycle test plus DNA reagent, apoptosis evaluation using FITC-Annexin V. Analyze using FACSCalibur flowcytometer. β HCG evaluation using Abbott AxSym total β-HCG reagent pack.
 Results: The group of cell that get ethynil estradiol in concentration 10 nmol/mL had cell proliferation index, amount cells and β HCG level higher than control after 72 hours observation. The group of cell that get desogestrel in concentration 100 nmol/mL have cell proliferation index, amount cells and β HCG level lower than control after 48 hours observation. There are no differences of apoptosis between the two group and control.
 Conclusion: Ethynil estradiol will increase proliferation of hydatidiform mole trophoblast cell, while desogestrel will decrease proliferation of hydatidiform mole trophoblast cell. There are no effect of ethynil estradiol and desogestrel on apoptosis of hydatidiform mole trophoblast cell.

Successful delivery of a twin pregnancy with complete hydatidiform mole and coexistent live fetus: a case report and review of literature

Background: A twin pregnancy consisting of either a complete or partial hydatidiform mole and a fetus is rare. The reported incidence ranges from 1:22,000–100,000 pregnancies, and complete hydatidiform mole with a coexistent fetus (CHMCF) comprises the majority of these cases. The management of CHMCF is controversial, as maternal risk with continuation of the pregnancy should be weighed against fetal survival. Women with CHMCF are at risk of developing preeclampsia, gestational diabetes, hyperthyroidism, antepartum hemorrhage, and gestational trophoblastic neoplasia. Case: We report a case of a healthy 32-year-old woman in her third pregnancy. She presented at 18 weeks gestation with vaginal bleeding and a significantly large uterus relative to the gestational age. Ultrasound showed CHMCF with a beta-hCG value of 398,800 IU/L. After careful discussion with the patient and after considering her options, she elected to continue the pregnancy. She was closely monitored for complications and had no maternal or fetal concerns. An elective cesarean delivery was performed at 32 weeks. A live female infant was delivered together with a normal placenta and a complete mole. The mother and baby were discharged in good condition after 2 days. A histopathological examination of the molar tissue confirmed the CHMCF diagnosis. No finding of gestational trophoblastic neoplasia (GTN) was discovered throughout one-year follow-up. Conclusion: Successful pregnancy outcomes can be achieved in cases of CHMCF. Comprehensive counseling with the patient regarding possible complications is important. Closely monitoring the mother for any complications and performing ongoing fetal surveillance are essential. Delivery should be planned at a tertiary center with good facilities and neonatal support.

Gingival Absorption of Rat Periodontal Disease Model and Rat Tooth Movement Model

In order to study the relationship between periodontal disease and alveolar bone resorption and the relationship between orthodontic treatment and alveolar bone resorption, two models of periodontal disease rats and tooth movement rats were constructed. The clinical diagnosis of gingival index, plaque index and depth of probing in the rat model of periodontal disease confirmed the successful construction of the periodontal disease model, and then the hematoxylin and eosin staining of the maxillary first molar tissue section revealed periodontal disease can cause aggregation of cementoclast and observation of nonstained sections showed that the area of absorption of periodontal disease rats was significantly increased compared with healthy rats. In this study, the rat model of tooth movement was also used. The left side of the maxillary teeth of the rat was injected with Ibanonate sodium phosphate in the experimental group, and the right side was injected with normal saline as the control group. It was found that in the tissue sections treated with Ibanonate sodium phosphate, the cementoclast were reduced and the degree of fang alveolar bone resorption was reduced. It indicated that the treatment has an inhibitory effect on fang alveolar bone resorption, which proved that Ibanonate sodium phosphate can be used in the treatment of orthodontic treatment of root resorption.

Gene Therapy Using Adeno-Associated Virus Serotype 8 Encoding TNAP-D10 Improves the Skeletal and Dentoalveolar Phenotypes in Alpl-/- Mice.

ABSTRACTHypophosphatasia (HPP) is caused by loss‐of‐function mutations in the ALPL gene that encodes tissue‐nonspecific alkaline phosphatase (TNAP), whose deficiency results in the accumulation of extracellular inorganic pyrophosphate (PPi), a potent mineralization inhibitor. Skeletal and dental hypomineralization characterizes HPP, with disease severity varying from life‐threatening perinatal or infantile forms to milder forms that manifest in adulthood or only affect the dentition. Enzyme replacement therapy (ERT) using mineral‐targeted recombinant TNAP (Strensiq/asfotase alfa) markedly improves the life span, skeletal phenotype, motor function, and quality of life of patients with HPP, though limitations of ERT include frequent injections due to a short elimination half‐life of 2.28 days and injection site reactions. We tested the efficacy of a single intramuscular administration of adeno‐associated virus 8 (AAV8) encoding TNAP‐D10 to increase the life span and improve the skeletal and dentoalveolar phenotypes in TNAP knockout (Alpl −/−) mice, a murine model for severe infantile HPP. Alpl −/− mice received 3 × 1011 vector genomes/body of AAV8‐TNAP‐D10 within 5 days postnatal (dpn). AAV8‐TNAP‐D10 elevated serum ALP activity and suppressed plasma PPi. Treatment extended life span of Alpl −/− mice, and no ectopic calcifications were observed in the kidneys, aorta, coronary arteries, or brain in the 70 dpn observational window. Treated Alpl −/− mice did not show signs of rickets, including bowing of long bones, enlargement of epiphyses, or fractures. Bone microstructure of treated Alpl −/− mice was similar to wild type, with a few persistent small cortical and trabecular defects. Histology showed no measurable osteoid accumulation but reduced bone volume fraction in treated Alpl −/− mice versus controls. Treated Alpl −/− mice featured normal molar and incisor dentoalveolar tissues, with the exceptions of slightly reduced molar enamel and alveolar bone density. Histology showed the presence of cementum and normal periodontal ligament attachment. These results support gene therapy as a promising alternative to ERT for the treatment of HPP. © 2021 The Authors. Journal of Bone and Mineral Research published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research (ASBMR).

Therapeutic approach to the iatrogenic invasive mole: A report of two cases

Introduction. Invasive mole, a form of gestational trophoblastic neoplasia (GTN), is defined as penetration of molar tissue into the myometrium and/or presence of extra-uterine metastases. An invasive mole arising from a complete hydatidiform mole is more common than an invasive mole arising from the partial hydatidiform mole. Dilatation and uterine evacuation and/or curettage (D&E/C) is the first step in managing molar pregnancy. Uterine perforation is the most serious complication of this procedure. A less common one is the false passage. Case report. The first case report describes a 47-year-old woman who was referred to our Clinic under the suspicion of GTN, with elevated serum beta human chorionic gonadotropin (beta hCG) levels. Intraoperatively, devitalized ovular tissue arising from the uterine perforation was observed. Histopathological exam (HPE) of tissue obtained from hysterectomy confirmed an invasive mole as a result of uterine perforation made during D&E/C. The second patient, a 32-year-old woman with vaginal bleeding, nausea, and high levels of serum beta hCG levels was admitted to our Clinic. After four D&E/C, and persistently high levels of serum beta hCG levels, explorative laparotomy has been performed. A false passage created during D&E/C with necrotic and molar tissue was observed. The partial hydatidiform mole was confirmed by the HPE. Conclusion. This is the second reported case in the literature of postmolar GTN as a result of iatrogenic perforation of the uterus, and also the first described case of postmolar GTN arising from a false passage, created as an iatrogenic complication of D&E/C. A nation-al survey of iatrogenic events during the treatment of gestational trophoblastic disaeses is needed since these events could completely change the therapeutic strategies in managing these diseases.

MiR-30a-5p inhibits proliferation and metastasis of hydatidiform mole by regulating B3GNT5 through ERK/AKT pathways.

Hydatidiform moles are gestational trophoblastic disease. They are abnormal proliferations of trophoblast cells that have the potential to become cancerous. miR‐miR30a‐5p is a tumour suppressor that participates in the development of numerous diseases. However, the role of miR‐30a in hydatidiform moles and the mechanisms underlying its effects are presently unclear. This study explored the levels of miR‐30a and B3GNT5 expression in human hydatidiform mole tissue. The results showed that miR‐30a and B3GNT5 were differentially expressed in normal placenta and hydatidiform mole, and miR‐30a decreased cell proliferation, invasion and migration in trophoblast cell lines. Upon further examination, it was confirmed that miR‐30a directly targeted the 3’untranslated region of B3GNT5 using a dual‐luciferase assay. The results of the present study also revealed that miR‐30a reduced the proliferation, invasion and migration ability in JAR and BeWo cells by regulating B3GNT5, which may inactivate the ERK and AKT signalling pathways. This study demonstrated that miR‐30a was a novel target B3GNT5 that serves an important role in the development of hydatidiform moles, suggesting that miR‐30a may serve as a novel potential biomarker or useful diagnostic and therapeutic tool for hydatidiform moles in clinical settings.

ABO blood type compatibility is not a risk factor for gestationaltrophoblastic neoplasia development from androgenetic completehydatidiform moles.

Complete hydatidiform moles (CHMs) are allografts to patients in terms of an androgenetic origin. Thus, some immunological reactions may be involved in the development of gestational trophoblastic neoplasia (GTN) from CHMs. This study aimed to evaluate the effect of ABO blood group on the prognosis of androgenetic CHMs. A total of 129 patients who were diagnosed as having CHMs based on multiplex short tandem repeat polymorphism analysis were included. The ABO blood types of molar tissues were determined by single-nucleotide polymorphisms in the ABO gene using a high-resolution melting assay. The incidence of GTN was compared based on ABO compatibility between the patients and their molar tissues. The overall incidence of GTN was 17.1% (22/129). Gestational trophoblastic neoplasia occurred in 10.8% (4/37), 14.8% (8/54), 22.2% (6/27), and 36.4% (4/11) of type O, A, B, and AB patients, respectively. Type AB patients tended to develop GTN compared with other blood type patients (P=.093). In ABO type of CHMs, GTN occurrence was not significantly different as it was 16.4% (10/64), 16.0% (8/50), and 22.2% (4/18) for types O, A, and B, respectively (P=.854). According to the ABO incompatibility between patients and molar tissues, GTN occurred in 19.1% (18/94) of the compatible cases and 11.4% (4/35) of the incompatible cases; the occurrence was not significantly different (P=.223). Patients with type AB tended to develop GTN. However, ABO compatibility between patients and molar tissues had no relationship with GTN occurrence.

Complete Hydatidiform Mole Coexisting Two Live Fetuses in Triplet Pregnancy

We introduce a case of triplet pregnancy with a complete hydatidiform mole coexisting two viable fetuses and important issues for prenatal care in both mothers and fetuses due to a significantly high risk for both mothers and fetuses. A 32-year-old patient conceived a triplet pregnancy after intrauterine insemination. At 14 weeks of gestation, ultrasonography revealed hydatidiform moles that were adjacent to two fetuses. Fetal karyotyping of the two fetuses showed normal results. The second baby developed fetal growth restriction at 24 weeks of gestation and preeclampsia developed at 29 weeks of gestation. At 31 weeks and one day of gestation, cesarean section was performed due to fetal distress of the second baby. Microscopically, the molar tissue was favorably diagnosed as complete hydatidiform mole. This is successful delivery report of a pregnant woman who had triplet pregnancy complicated with one complete hydatidiform mole coexisting two viable fetuses in Korea.

Comprehensive analysis of 204 sporadic hydatidiform moles: revisiting risk factors and their correlations with the molar genotypes.

Hydatidiform mole (HM) is an aberrant human pregnancy characterized by excessive trophoblastic proliferation and abnormal embryonic development. HM has two morphological types, complete (CHM) and partial (PHM), and non-recurrent ones have three genotypic types, androgenetic monospermic, androgenetic dispermic, and triploid dispermic. Most available studies on risk factors predisposing to different types of HM and their malignant transformation mainly suffer from the lack of comprehensive genotypic analysis of large cohorts of molar tissues combined with accurate postmolar hCG follow-up. Moreover, 10-20% of patients with one HM have at least one non-molar miscarriage, which is higher than the frequency of two pregnancy losses in the general population (2-5%), suggesting a common genetic susceptibility to HM and miscarriages. However, the underlying causes of the miscarriages in these patients are unknown. Here, we comprehensively analyzed 204 HM, mostly from patients referred to the Quebec Registry of Trophoblastic Diseases and for which postmolar hCG monitoring is available, and 30 of their non-molar miscarriages. We revisited the risk of maternal age and neoplastic transformation across the different HM genotypic categories and investigated the presence of chromosomal abnormalities in their non-molar miscarriages. We confirm that androgenetic CHM is more prone to gestational trophoblastic neoplasia (GTN) than triploid dispermic PHM, and androgenetic dispermic CHM is more prone to high-risk GTN and choriocarcinoma (CC) than androgenetic monospermic CHM. We also confirm the association between increased maternal age and androgenetic CHM and their malignancies. Most importantly, we demonstrate for the first time that patients with an HM and miscarriages are at higher risk for aneuploid miscarriages [83.3%, 95% confidence interval (CI): 0.653-0.944] than women with sporadic (51.5%, 95% CI: 50.3-52.7%, p value = 0.0003828) or recurrent miscarriages (43.8%, 95% CI: 40.7-47.0%, p value = 0.00002). Our data suggest common genetic female germline defects predisposing to HM and aneuploid non-molar miscarriages in some patients.

A KHDC3L mutation resulting in recurrent hydatidiform mole causes genome-wide DNA methylation loss in oocytes and persistent imprinting defects post-fertilisation

BackgroundMaternal effect mutations in the components of the subcortical maternal complex (SCMC) of the human oocyte can cause early embryonic failure, gestational abnormalities and recurrent pregnancy loss. Enigmatically, they are also associated with DNA methylation abnormalities at imprinted genes in conceptuses: in the devastating gestational abnormality biparental complete hydatidiform mole (BiCHM) or in multi-locus imprinting disease (MLID). However, the developmental timing, genomic extent and mechanistic basis of these imprinting defects are unknown. The rarity of these disorders and the possibility that methylation defects originate in oocytes have made these questions very challenging to address.MethodsSingle-cell bisulphite sequencing (scBS-seq) was used to assess methylation in oocytes from a patient with BiCHM identified to be homozygous for an inactivating mutation in the human SCMC component KHDC3L. Genome-wide methylation analysis of a preimplantation embryo and molar tissue from the same patient was also performed.ResultsHigh-coverage scBS-seq libraries were obtained from five KHDC3Lc.1A>G oocytes, which revealed a genome-wide deficit of DNA methylation compared with normal human oocytes. Importantly, germline differentially methylated regions (gDMRs) of imprinted genes were affected similarly to other sequence features that normally become methylated in oocytes, indicating no selectivity towards imprinted genes. A range of methylation losses was observed across genomic features, including gDMRs, indicating variable sensitivity to defects in the SCMC. Genome-wide analysis of a pre-implantation embryo and molar tissue from the same patient showed that following fertilisation methylation defects at imprinted genes persist, while most non-imprinted regions of the genome recover near-normal methylation post-implantation.ConclusionsWe show for the first time that the integrity of the SCMC is essential for de novo methylation in the female germline. These findings have important implications for understanding the role of the SCMC in DNA methylation and for the origin of imprinting defects, for counselling affected families, and will help inform future therapeutic approaches.

Microsatellite DNA Genotyping and Flow Cytometry Ploidy Analyses of Formalin-fixed Paraffin-embedded Hydatidiform Molar Tissues

Hydatidiform mole (HM) is an abnormal human pregnancy characterized by excessive trophoblastic proliferation and abnormal embryonic development. There are two types of HM based on microscopic morphological evaluation, complete HM (CHM) and partial HM (PHM). These can be further subdivided based on the parental contribution to the molar genomes. Such characterization of HM, by morphology and genotype analyses, is crucial for patient management and for the fundamental understanding of this intriguing pathology. It is well documented that morphological analysis of HM is subject to wide interobserver variability and is not sufficient on its own to accurately classify HM into CHM and PHM and distinguish them from hydropic non-molar abortions. Genotyping analysis is mostly performed on DNA and tissues from formalin-fixed paraffin-embedded (FFPE) products of conception, which have less than optimal quality and may consequently lead to wrong conclusions. In this article, detailed protocols for multiplex genotyping and flow cytometry analyses of FFPE molar tissues are provided, along with the interpretation of the results of these methods, their troubleshooting, and integration with the morphological evaluation, p57KIP2 immunohistochemistry, and fluorescence in situ hybridization (FISH) to reach a correct and robust diagnosis. Here, the authors share the methods and lessons learned in the past 10 years from the analysis of approximately 400 products of conception.

2,4-D causes oxidative stress induction and apoptosis in human dental pulp stem cells (hDPSCs).

2,4-Dicholorophenoxy acetic acid (2,4-D) is a worldwide used hormone herbicide. Human dental pulp stem cells (hDPSCs) as a potential source of mesenchymal stem cells provide a confident model system for the assessments of chemicals in vitro. The main objective of this study was to examine the biological effects and damages attributed to 2,4-D on hDPSCs. hDPSCs were isolated from third molar pulp tissues and their mesenchymal identity were evaluated. Then, hDPSCs were treated with increasing concentrations of 2,4-D (0.1μM-10mM). Cell viability assay and cumulative cell counting were carried out to address 2,4-D effects on biological parameters of hDPSCs. Cell cycle distribution, ROS level and ALP activity were measured before and after treatment. AO/EB staining and caspase 3/7 activity were investigated to detect the possible mechanisms of cell death. Flow-cytometric immunophenotyping and differentiation data confirmed the mesenchymal identity of cultivated hDPSCs. 2,4-D treatment caused a hormetic response in the viability and growth rate of hDPSCs. G0/G1 cell cycle arrest, enhanced ROS level, and reduced ALP activity were detected in hDPSCs treated with EC50 dose of 2,4-D. AO/EB staining showed a higher percentage of alive cells in lower concentrations of the herbicide. The increment in 2,4-D dose and the number of early and late apoptotic cells were increased. DAPI staining and caspase 3/7 assay validated the induction of apoptosis. 2,4-D concentrations up to 100μM did not affect hDPSCs viability and proliferation. The intense cellular oxidative stress and apoptosis were observed at higher concentration.

The effect of ethynyl estradiol and desogestrel on proliferation and apoptosis hydatidiform mole trophoblast cell

This study purpose to explore the effect of ethynyl estradiol and desogestrel on proliferation and apoptosis hydatidiform mole trophoblast cell. Hydatidiform mole tissue, trophoblast cells were isolated and culture at RPMI 20% FBS medium. Trophoblast cells divided into three groups observation. First group get ethynyl estradiol 10 nmol/mL, second group get desogestrel 100 nmol/mL, third group get DMSO 1%. Cells incubated and observe at 24, 48, 72, 96 hours. Cell cycle, apoptosis and β HCG was evaluated. The group of cells that get ethynyl estradiol in concentration 10 nmol/mL had cell proliferation index, amount cells and β HCG level higher than control after 72 hours observation. The group of cells that get desogestrel in concentration 100 nmol/mL have cell proliferation index, amount cells and β HCG level lower than control after 48 hours observation. There are no differences of apoptosis between the two group and control. Conclusion: Ethynyl estradiol will increase proliferation of hydatidiform mole trophoblast cell, while desogestrel will decrease proliferation of hydatidiform mole trophoblast cell. There is no effect of ethynyl estradiol and desogestrel on apoptosis of hydatidiform mole trophoblast cell.

Surgical reintervention in hemifacial microssomy

Background: The Hemifacial Microsomia (HFM) is a progressive deformity of the skeleton and mole tissue, thus, as a variable that results in hypoplasia of the affected structures, being the first manifestations in the mandible.

NLRP7 and KHDC3L variants in Chinese patients with recurrent hydatidiform moles.

Recurrent hydatidiform moles are reportedly biparental complete moles and related to mutated NLRP7 and KHDC3L. This study was designed to identify mutations of gene NLRP7 and KHDC3L in biparental complete moles. In this study, we have screened NLRP7 and KHDC3L mutations in five patients with recurrent moles and five with sporadic moles. Molar tissues and blood samples were collected from patients and their partners. Genotypes of the molar tissues were determined based on short tandem repeat polymorphism. The coding exons of NLRP7 and KHDC3L were sequenced. Two patients with recurrent moles had biparental complete moles, while all other patients had androgenetic complete moles. Three non-synonymous variants in NLRP7 (c.955 G>A, c.1280 T>C and c.1441 G>A) and one in KHDC3L (c.602 C>G) were identified in patients with recurrent moles. NLRP7 c.1441 G>A and c.1280 T>C were mutations found in the Chinese population, while c.1441 G>A was only detected in patients with biparental complete moles in this study. Genotyping can be used to differentiate biparental complete moles from androgenetic moles and to predict the risk of recurrent moles in future pregnancies. NLRP7 c.1441 G>A may associate with biparental complete moles. Biparental complete moles exhibit genetic heterogeneity.

Proteomic identification of predictive biomarkers for malignant transformation in complete hydatidiform moles

IntroductionProtein expression in cells are associated with oncogenesis. This study aims to explore proteomic profiles and discover potential biomarkers that can predict malignant transformation of hydatidiform mole. MethodsRetrospective analysis was done in 14 cases of remission hydatidiform mole and 14 cases of hydatidiform mole who later developed malignancy (GTN group). Molar tissues were retrieved from −70 °C frozen tissue. Subsequently, a large-scale proteomic analysis was performed to identify proteins and compare their abundance levels in the preserved molar tissues from these two groups using a dimethyl-labeling technique coupled with liquid chromatography-tandem mass spectrometry (LC-MS/MS). ResultsA total of 2,153 proteins were identified from all samples. 22 and 10 proteins were significantly up-regulated and down-regulated, respectively, in the GTN group compared with the mole group. These altered proteins were found in several biological groups such as cell-cell adhesion, secreted proteins, and ribonucleoproteins. Several hormone-related proteins were among the most up-regulated proteins in the GTN group including choriogonadotropin subunit beta (β-hCG) and alpha (α-hCG), growth/differentiation factor 15, as well as both pregnancy-specific beta-1-glycoproteins 2 and 3. In contrast, protein S100-A11 and l-lactate dehydrogenase A chain, were down-regulated in molar tissue from most patients in the GTN group. DiscussionThis study identified a set of differentially expressed proteins in molar tissues that could potentially be further examined as predictive biomarkers for the malignant transformation of CHMs. A molar proteome database was constructed and can be accessible online at http://sysbio.chula.ac.th/Database/GTD_DB/Supplementary_Data.xlsx.

The genetics of recurrent hydatidiform moles: new insights and lessons from a comprehensive analysis of 113 patients

AbstractHydatidiform mole is an aberrant human pregnancy characterized by early embryonic arrest and excessive trophoblastic proliferation. Recurrent hydatidiform moles are defined by the occurrence of at least two hydatidiform moles in the same patient. Fifty to eighty percent of patients with recurrent hydatidiform moles have biallelic pathogenic variants in NLRP7 or KHDC3L. However, in the remaining patients, the genotypic types of the moles are unknown. We characterized 80 new hydatidiform mole tissues, 57 of which were from patients with no mutations in the known genes, and we reviewed the genotypes of a total of 123 molar tissues. We also reviewed mutation analysis in 113 patients with recurrent hydatidiform moles. While all hydatidiform moles from patients with biallelic NLRP7 or KHDC3L mutations are diploid biparental, we demonstrate that those from patients without mutations are highly heterogeneous and only a small minority of them are diploid biparental (8%). The other mechanisms that were found to recur in patients without mutations are diploid androgenetic monospermic (24%) and triploid dispermic (32%); the remaining hydatidiform moles were misdiagnosed as moles due to errors in the analyses and/or their unusual mechanisms. We compared three parameters of genetic susceptibility in patients with and without mutations and show that patients without mutations are mostly from non-familial cases, have fewer reproductive losses, and more live births. Our data demonstrate that patients with recurrent hydatidiform moles and no mutations in the known genes are, in general, different from those with mutations; they have a milder genetic susceptibility and/or a multifactorial etiology underlying their recurrent hydatidiform moles. Categorizing these patients according to the genotypic types of their recurrent hydatidiform moles may facilitate the identification of novel genes for this entity.

Specificity Protein 7 Is Required for Proliferation and Differentiation of Ameloblasts and Odontoblasts.

The Sp7/Osterix transcription factor is essential for bone development. Mutations of the Sp7 gene in humans are associated with craniofacial anomalies and osteogenesis imperfecta. However, the role of Sp7 in embryonic tooth development remains unknown. Here we identified the functional requirement of Sp7 for dentin synthesis and tooth development. Sp7-null mice exhibit craniofacial dysmorphogenesis and are completely void of alveolar bone. Surprisingly, initial tooth morphogenesis progressed normally in Sp7-null mice. Thus the formation of alveolar bone is not a prerequisite for tooth morphogenesis. Sp7 is required for mineralization of palatal tissue but is not essential for palatal fusion. The reduced proliferative capacity of Sp7-deficient ectomesenchyme results in small and misshapen teeth with randomly arranged cuboidal preodontoblasts and preameloblasts. Sp7 promotes functional maturation and polarization of odontoblasts. Markers of mature odontoblast (Col1a, Oc, Dspp, Dmp1) and ameloblast (Enam, Amelx, Mmp20, Amtn, Klk4) are barely expressed in incisors and molar tissues of Sp7-null mice. Consequently, dentin and enamel matrix are absent in the Sp7-null littermates. Interestingly, the Sp7 expression is restricted to cells of the dental mesenchyme indicating the effect on oral epithelium-derived ameloblasts is cell-nonautonomous. Abundant expression of Fgf3 and Fgf8 ligand was noted in the developing tooth of wild-type mice. Both ligands were remarkably absent in the Sp7-null incisor and molar, suggesting cross-signaling between mesenchyme and epithelium is disrupted. Finally, promoter-reporter assays revealed that Sp7 directly controls the expression of Fgf-ligands. Together, our data demonstrate that Sp7 is obligatory for the differentiation of both ameloblasts and odontoblasts but not for the initial tooth morphogenesis. © 2018 American Society for Bone and Mineral Research.

Management of bilaterally immature permanent teeth using various treatment modalities

Management of immature permanent teeth exhibiting symptomatic or necrotic pulp is clinically challenging. Appropriate diagnosis and case selection, and good management, ensure good outcomes. Here, the management of the bilateral, immature, permanent mandibular first molars of a 5.6-year-old male who exhibited reversible pulpitis of the mandibular first right molar (tooth #46) and normal apical tissue and pulpal necrosis with asymptomatic apical periodontitis of the mandibular first left molar (#36) is presented. The former tooth was managed using vital pulp therapy (pulpotomy) and the latter employing revascularization. Follow-up examinations revealed complete formation of the mesial and distal roots of tooth #46, complete formation of the mesial root, but incomplete formation of the distal root of tooth #36, with radiolucency evident within the root.