Hypocalcified Amelogenesis Imperfecta Research Articles

A multidisciplinary approach for the diagnosis of hypocalcified amelogenesis imperfecta in two Chilean families

Objective. The purpose of this study was to conduct a multidisciplinary analysis of a specific type of tooth enamel disturbance (amelogenesis imperfecta) affecting two Chilean families to obtain a precise diagnosis and to investigate possible underlying mutations. Materials and methods. Two non-related families affected with amelogenesis imperfecta were evaluated with clinical, radiographic and histopathological methods. Furthermore, pedigrees of both families were constructed and the presence of eight mutations in the enamelin gene (ENAM) and three mutations in the enamelysin gene (MMP-20) were investigated by PCR and direct sequencing. Results. In the two affected patients, the dental malformation presented as soft and easily disintegrated enamel and exposed dark dentin. Neither of the affected individuals presented with a dental and skeletal open bite. Histologically, a high level of an organic matrix with prismatic organization was found. Genetic analysis indicated that the condition is autosomal recessive in one family and either autosomal recessive or due to a new mutation in the other family. Molecular mutational analysis revealed that none of the eight mutations previously described in the ENAM gene or the three mutations in the MMP-20 gene were present in the probands. Conclusion. A multidisciplinary analysis allowed for a diagnosis of hypocalcified amelogenesis imperfecta, Witkop type III, which was unrelated to previously described mutations in the ENAM or MMP-20 genes.

Hardness and microshear bond strength to enamel and dentin of permanent teeth with hypocalcified amelogenesis imperfecta

Adhesive procedures are often required to restore teeth affected by hypocalcified amelogenesis imperfecta (HAI). To evaluate the hardness of enamel/dentin of teeth affected by HAI and the bond strength to these substrates, as well the influence of 5% NaOCl on bond strength. Permanent molars presenting HAI and sound third molars were used. Enamel surfaces were wet-flattened and Knoop hardness was assessed. The two-step, etch-and-rinse adhesive Single Bond 2 (3M ESPE) was applied and resin cylinders bonded to the surfaces and submitted to microshear testing. The subjacent medium dentin was then exposed by wet-grinding. Hardness readings and microshear testing were carried out again. The relationship between hardness and bond strength was assessed by nonlinear regression analysis. Hardness of normal enamel was higher than hardness of enamel affected by HAI, whereas dentin hardness did not differ from normal to HAI-affected teeth. Enamel and dentin hardness were similar for teeth affected by HAI. Higher bond strengths were obtained to the normal tooth tissues. Dentin bond strength was higher than enamel bond strength. NaOCl exposure did not influence bond strengths. A positive linear relationship between enamel hardness and bond strength was observed. HAI imposes challenges to bonding to enamel and dentin.

Ultrastructural Analyses of Deciduous Teeth Affected by Hypocalcified Amelogenesis Imperfecta from a Family with a Novel Y458X FAM83H Nonsense Mutation

Background: Nonsense mutations in FAM83H are a recently described underlying cause of autosomal dominant (AD) hypocalcified amelogenesis imperfecta (AI). Objective: This study aims to report a novel c.1374C>A p.Y458X nonsense mutation and describe the associated ultrastructural phenotype of deciduous teeth. Methods: A family of European origin from the Iberian Peninsula with AD-inherited AI was ascertained. Family members were assessed through clinical examination and supporting investigations. Naturally exfoliated deciduous teeth from 2 siblings were investigated by scanning electron microscopy (SEM), energy dispersive X-ray analysis (EDX) and transverse microradiography (TMR). Results: On clinical and radiographic investigation the appearances of the affected deciduous and permanent teeth were consistent with hypocalcified AI with small focal areas of more normal looking enamel. DNA sequencing identified a novel c.1374C>A p.Y458X FAM83H nonsense mutation in affected, but not in either unaffected family members or unrelated controls. Exfoliated teeth were characterised by substantial post-eruptive enamel loss on gross examination. Irregular, poor quality enamel prisms were observed on SEM. These were coated in amorphous material. TMR and EDX confirmed reduced mineral and increased organic content in enamel, respectively. Conclusions:FAM83H nonsense mutations have recently been recognised as a cause of AD hypocalcified AI. We report a novel nonsense FAM83H mutation and describe the associated preliminary ultrastructural phenotype in deciduous teeth. This is characterised by poorly formed enamel rods with inappropriate retention of amorphous material, which is likely to represent retained organic matrix that contributes to the overall hypomineralised phenotype.

Identification of a novel FAM83H mutation and microhardness of an affected molar in autosomal dominant hypocalcified amelogenesis imperfecta

To determine the underlying molecular genetic aetiology of a family with the hypocalcified form of amelogenesis imperfecta and to investigate the hardness of the enamel and dentine of a known FAM83H mutation. Mutational screening of the FAM83H on the basis of candidate gene approach was performed. All exons and exon-intron boundaries was amplified and sequenced. A microhardness test was performed to measure the Vickers microhardness value. A novel nonsense mutation (c.1354C>T, p.Q452X) was identified in the last exon of FAM83H, which resulted in soft, uncalcified enamel. The affected enamel was extremely soft (about 17% of the normal control), but the underlying dentine was as hard as the normal control. Mutational analysis revealed a novel mutation in FAM83H gene. Hardness of dentine was not affected by the mutation, whilst the enamel was extremely soft.

Novel FAM83H mutations in Turkish families with autosomal dominant hypocalcified amelogenesis imperfecta

Novel <i>FAM83H </i>mutations in Turkish families with autosomal dominant hypocalcified amelogenesis imperfecta

Mutational spectrum of FAM83H: the C-terminal portion is required for tooth enamel calcification.

Dental enamel forms through the concerted activities of specialized extracellular matrix proteins, including amelogenin, enamelin, MMP20, and KLK4. Defects in the genes encoding these proteins cause non-syndromic inherited enamel malformations collectively designated as amelogenesis imperfecta (AI). These genes, however, account for only about a quarter of all AI cases. Recently we identified mutations in FAM83H that caused autosomal dominant hypocalcified amelogenesis imperfecta (ADHCAI). Unlike other genes that cause AI, FAM83 H does not encode an extracellular matrix protein. Its location inside the cell is completely unknown, as is its function. We here report novel FAM83H mutations in four kindreds with ADHCAI. All are nonsense mutations in the last exon (c.1243G>T, p.E415X; c.891T>A, p.Y297X; c.1380G>A, p.W460X; and c.2029C>T, p.Q677X). These mutations delete between 503 and 883 amino acids from the C-terminus of a protein normally comprised of 1179 residues. The reason these mutations cause such extreme defects in the enamel layer without affecting other parts of the body is not known yet. However it seems evident that the large C-terminal part of the protein is essential for proper enamel calcification.

FAM83H Mutations in Families with Autosomal-Dominant Hypocalcified Amelogenesis Imperfecta

Amelogenesis imperfecta (AI) is a collection of diverse inherited disorders featuring dental-enamel defects in the absence of significant nondental symptoms. AI phenotypes vary and are categorized as hypoplastic, hypocalcified, and hypomaturation types. Phenotypic specificity to enamel has focused research on genes encoding enamel-matrix proteins. We studied two families with autosomal-dominant hypocalcified AI and have identified nonsense mutations (R325X and Q398X) in the FAM83H gene on chromosome 8q24.3. The mutations perfectly cosegregate with the disease phenotype and demonstrate that FAM83H is required for proper dental-enamel calcification.

Effect of deproteinization on composite bond strength in hypocalcified amelogenesis imperfecta

The aim of this study was to evaluate the effect of the treatment of sodium hypochlorite (NaOCl) after acid conditioning of the enamel and dentin of the primary teeth affected with hypocalcified amelogenesis imperfecta (HCAI) on the shear bond strength of the composite material. Primary teeth from a 12-year-old girl affected with HCAI and primary teeth collected from apparently healthy children were used. A total of four groups, experimental and control with and without NaOCl treatment were specified. In the control group conventional composite procedure was performed and in the treatment group 5% NaOCl was applied after acid conditioning and then the procedure continued as in the control group. In teeth affected with HCAI, enamel shear bond strengths were significantly enhanced in the treatment group compared with the conventional procedure. Deproteinization could be attributed as effective in enhancing the enamel bonding in HCAI teeth and could be used to overcome the high failure rates of adhesive restorations in HCAI cases.

Dentine structure and mineralization in hypocalcified amelogenesis imperfecta: a quantitative X-ray histochemical study.

This study was undertaken in order to establish the structural and mineralization pattern of the response of dentine to alterations in enamel in hypocalcified amelogenesis imperfecta (AI). The images and data obtained with scanning electron microscopy and electron probe X-ray microanalysis in enamel and dentine specimens from control and affected teeth were compared in this study. We compared 46 fragments of permanent teeth from patients with clinically diagnosed hypocalcified AI and 20 normal permanent teeth. All specimens were prepared for electron probe X-ray microanalysis. Dentine is characterized by thickening of the peritubular dentine and partial obliteration of the dentinal tubules that does not give rise to a compact sclerotic cast. In dentine, calcium levels were significantly higher in teeth with clinically hypocalcified AI in relation with control teeth (P < 0.001). Dentine is affected in hypocalcified AI increasing mineralization (narrower tubules and higher content of calcium) in response to enamel disorder.

Exclusion of candidate genes in two families with autosomal dominant hypocalcified amelogenesis imperfecta.

The amelogenesis imperfectas (AI) are a group of hereditary enamel defects characterized by clinical and genetic diversity. The most common AI types are inherited as autosomal traits. Three mutations of the enamelin (ENAM) gene have been found in cases of autosomal dominant hypoplastic AI. The gene(s) responsible for hypocalcified forms of AI have not been identified, although a number of autosomal genes have been proposed as candidates for AI based on their expression by ameloblasts, including ameloblastin and enamelin (chromosome 4q13.3), tuftelin (chromosome 1q21), enamelysin (chromosome 11q22.3-q23) and kallikrein 4 (chromosome 19q13.3-q13.4). To localize the gene(s) responsible for autosomal dominant hypocalcified AI, we evaluated support for/against linkage of AI to genetic markers spanning five AI candidate genes in two extended families. Our data excluded all proposed candidate gene regions as causal for autosomal dominant hypocalcified AI in these families. These linkage findings provide further evidence for genetic heterogeneity among families with autosomal dominant AI and indicate that, at least, some forms of autosomal dominant hypocalcified AI are not caused by a gene in the five most commonly reported AI candidate genes.

Immunochemical and biochemical characteristics of enamel proteins in hypocalcified amelogenesis imperfecta

Amelogenesis imperfecta is a hereditary disease of the enamel that is unassociated with generalized defects. Cases of the condition are clinically classified into three groups: hypoplastic, hypomaturation, and hypocalcified. In this study, soluble protein fractions of the enamel from three patients with hypocalcified amelogenesis imperfecta were examined through the use of immunochemical and biochemical techniques. In immunochemical analyses done with a polyclonal anti-amelogenin antibody, all samples from enamel in which there was amelogenesis imperfecta were found to contain considerable amounts of amelogenin peptides. When an enamel sample from one patient was examined by Western-blot transfer and immunobinding analysis, the amelogenin fraction was found to consist of a 26-kDa molecule thought to be normally present in the outer layer of secretory-stage enamel. This enamel was also found to contain albumin as one of the major constituents of the protein fraction. These results suggest that hypocalcified amelogenesis imperfecta may in part be caused by a disturbance in matrix protein degradation during the maturation phase.

The enamel proteins in human amelogenesis imperfecta

Amelogenesis imperfecta comprises a unique group of hereditary conditions that result in abnormal enamel development. The purpose of this study was to characterize the enamel proteins in different amelogenesis imperfecta types and to determine if amelogenin, the principal matrix protein in normal developing enamel, was retained. Primary and/or permanent amelogenesis imperfecta teeth were analysed from 11 individuals. Normal teeth served as controls. Thin sections were cut with a diamond blade and enamel was dissected for analysis. The enamel proteins were characterized by amino acid analysis, sodium dodecyl sulphate-polyacrylamide gel electrophoresis, and Western blot analysis using; antiamelogenin antibodies. An increased protein content was seen in all hypocalcified and hypomaturation amelogenesis imperfecta cases. A slightly increased protein content was seen in two of four hypoplastic amelogenesis imperfecta cases. The enamel protein amino acid composition varied between the different amelogenesis imperfecta types. All three cases of hypomaturation amelogenesis imperfecta enamel showed an increased proline content compared with normal enamel or other amelogenesis imperfecta types. Hypocalcified amelogenesis imperfecta enamel had an increased tyrosine content while the other amino acids were generally similar in amount to normal enamel. Fully developed hypomaturation and hypocalcified amelogenesis imperfecta enamel showed cross-reactivity to antiamelogenin antibodies while normal enamel did not. Although both amelogenesis imperfecta types showed cross-reactivity, the banding patterns on Western blot analyses were markedly different. This investigation provides additional evidence that abnormal post-secretory processing of amelogenin is involved in hypomaturation and hypocalcified amelogenesis imperfecta. Furthermore, these results indicate that amelogenin retention can occur in a variety of amelogenesis imperfecta types. The unique amino acid compositions and distinct enamel protein species seen by electrophoresis and Western blot analyses suggest that different developmental processes might be involved in hypomaturation and hypocalcified amelogenesis imperfecta.