Changes In Ameloblasts Research Articles

The phosphorylation of serine55 in enamelin is essential for murine amelogenesis

Amelogenesis imperfecta (AI) is an inherited developmental enamel defect affecting tooth masticatory function, esthetic appearance, and the well-being of patients. As one of the major enamel matrix proteins (EMPs), enamelin (ENAM) has three serines located in Ser-x-Glu (S-x-E) motifs, which are potential phosphorylation sites for the Golgi casein kinase FAM20C. Defects in FAM20C have similarly been associated with AI. In our previous study of EnamRgsc514 mice, the Glu57 in the S55-X56-E57 motif was mutated into Gly, which was expected to cause a phosphorylation failure of Ser55 because Ser55 cannot be recognized by FAM20C. The severe enamel defects in ENAMRgsc514 mice reminiscent of Enam-knockout mouse enamel suggested a potentially important role of Ser55 phosphorylation in ENAM function. However, the enamel defects and ENAM dysfunction may also be attributed to distinct physicochemical differences between Glu57 and Gly57. To clarify the significance of Ser55 phosphorylation to ENAM function, we generated two lines of Enam knock-in mice using CRISPR-Cas9 method to eliminate or mimic the phosphorylation state of Ser55 by substituting it with Ala55 or Asp55 (designated as S55A or S55D), respectively. The teeth of 6-day or 4-week-old mice were subjected to histology, micro-CT, SEM, TEM, immunohistochemistry, and mass spectrometry analyses to characterize the morphological, microstructural and proteomic changes in ameloblasts, enamel matrix and enamel rods. Our results showed that the enamel formation and EMP expression in S55D heterozygotes (Het) were less disturbed than those in S55A heterozygotes, while both homozygotes (Homo) had no mature enamel formation. Proteomic analysis revealed alterations of enamel matrix biosynthetic and mineralization processes in S55A Hets. Our present findings indicate that Asp55 substitution partially mimics the phosphorylation state of Ser55 in ENAM. Ser55 phosphorylation is essential for ENAM function during amelogenesis.

Deletion of epithelial cell-specific p130Cas impairs the maturation stage of amelogenesis

Amelogenesis consists of secretory, transition, maturation, and post-maturation stages, and the morphological changes of ameloblasts at each stage are closely related to their function. p130 Crk-associated substrate (Cas) is a scaffold protein that modulates essential cellular processes, including cell adhesion, cytoskeletal changes, and polarization. The expression of p130Cas was observed from the secretory stage to the maturation stage in ameloblasts. Epithelial cell-specific p130Cas-deficient (p130CasΔepi-) mice exhibited enamel hypomineralization with chalk-like white mandibular incisors in young mice and attrition in aged mouse molars. A micro-computed tomography analysis and Vickers micro-hardness testing showed thinner enamel, lower enamel mineral density and hardness in p130CasΔepi- mice in comparison to p130Casflox/flox mice. Scanning electron microscopy, and an energy dispersive X-ray spectroscopy analysis indicated the disturbance of the enamel rod structure and lower Ca and P contents in p130CasΔepi- mice, respectively. The disorganized arrangement of ameloblasts, especially in the maturation stage, was observed in p130CasΔepi- mice. Furthermore, expression levels of enamel matrix proteins, such as amelogenin and ameloblastin in the secretory stage, and functional markers, such as alkaline phosphatase and iron accumulation, and Na+/Ca2++K+-exchanger in the maturation stage were reduced in p130CasΔepi- mice. These findings suggest that p130Cas plays important roles in amelogenesis (197 words).

Efficacy of transforming growth factor- β on development of ameloblasts and odontoblasts in tooth germ of young albino rat

Transforming growth factors were known as secreted factors that induced the growth of non-transformed cells. The study investigating the growth response of ameloblasts and odontoblasts to TGF-β. Four adult male and 12 adult female albino rats were used in this study. Mating took place between them. Female rats were divided into two groups each group consist of 6 rats: group I (control) rats were injected with 0.1 m1 phosphate buffer saline every two days from the first day till the labor time. Their offspring were considered the control group. Group II were injected intramuscular with TGF-β 40 ng dissolved in PBS every two days during the pregnancy period. Thirty off-springs rats of both groups were killed after 4 days of birth and the lower jaw of each was dissected. The sections were stained by hematoxyline and eosin and immuno for histological study. All the data will statistically be analyzed. Histological examination of tissue sections, showed differences in ameloblast and odontoblastic layers. Enamel matrix, dentin and predentin layers showed variable thickness when compared to the control group. While Immunohistochemical examination of both ameloblasts and odontoblasts of group II showed negative nuclear and positive cytoplasmic reaction of MMP-9 in all stained tissue sections. It was concluded that: within the utilized doses and experiment duration, TGF-β was associated with noticeable histological changes in ameloblasts and odontoblasts. TGF-β has a relatively mild stimulatory effect on the organic matrix secretory potential of odontoblasts was observed in dentin.

Ultrastructural and immunohistochemical study of the influence of fluoride excess on the development of rat incisor tooth buds

Little information is available on the pathogenesis of fluorosis during the fetal and initial postnatal period. In the present study, female rats received 0 (control), 7 or 100 ppm of sodium fluoride in drinking water, one week before breeding and throughout gestation and nursing periods. The hemimandibles of the offspring were collected at 0, 7 and 14 days of postnatal life (n = 5) and processed for morphological analyses by light and electron microscopy, immunohistochemical analysis for amelogenin and morphometric study of enamel matrix and ameloblasts of incisors. The results showed a decrease in matrix production at the secretory phase at all study periods for the 100 ppm group. In this same group, the secretory ameloblasts showed reduction of enamel matrix secretion, disorganization of mitochondrial crests, large vacuoles at the apical portion of the cytoplasm, retention of intracisternal material and dilatation of some cisterns in the rough endoplasmic reticulum. In the groups of animals aged 7 and 14 days, analysis of variance showed significant reduction (p<0.05) in cytoplasmic volume of 23.80% and 24.75%, respectively, in relation to the control group. The smooth-ended maturation ameloblasts exhibited a large number of vacuoles with electron-dense endocytic matrix, suggesting a delay in the resorption process. Immunohistochemical analysis showed no difference in the intensity and labeling pattern of the enamel matrix in any study group. Interestingly, in offspring at the age of 14 days for the 7 ppm group, there was an increase in the matrix length at the secretory phase. Therefore, part of the excessive dose of sodium fluoride given to the mother in drinking water can reach the offspring through the placenta and mother’s milk, causing morphological changes in ameloblasts and suggesting a reduction in secretion and a delay in matrix resorption.

Short exposure to high levels of fluoride induces stage‐dependent structural changes in ameloblasts and enamel mineralization

We tested the hypothesis that the sensitivity of forming dental enamel to fluoride (F-) is ameloblast developmental stage-dependent and that enamel mineralization disturbances at the surface of fluorotic enamel are caused by damage to late-secretory- and transitional-stage ameloblasts. Four-day-old hamsters received a single intraperitoneal dose of 2.5-20 mg NaF/kg body weight and were examined, 24 h later, by histology and histochemistry. A single dose of >or=5 mg of NaF/kg induced the formation of a hyper- followed by a hypomineralized band in the secretory enamel, without changing the ameloblast structure. At 10 mg of NaF/kg, cystic lesions became apparent under isolated populations of distorted late-secretory- and transitional-stage ameloblasts. Staining with von Kossa stain showed that the enamel under these lesions was hypermineralized. At 20 mg of NaF/kg, cystic lesions containing necrotic cells were also found in the early stages of secretory amelogenesis and were also accompanied with hypermineralization of the enamel surface. We concluded that the sensitivity to F- is ameloblast developmental stage-dependent. Groups of transitional ameloblasts are most sensitive, followed by those at early secretory stages. These data suggest that a F-induced increase in cell death in the transitional-stage ameloblasts accompanies the formation of cystic lesions, which may explain the formation of enamel pits seen clinically in erupted teeth.

Developmental disturbances of the rat molar induced by two diphosphonates.

Very few reports have been published about the effects of diphosphonates on the cells and tissues of developing teeth. The present study was designed to investigate possible morphological changes in ameloblasts and odontoblasts and relate these changes to defects in the enamel surface of erupted teeth. Young rats were injected subcutaneously with single or multiple doses of HEDP or Cl2MDP (10 mg P/kg b.w.). Light microscopic examination of developing maxillary first molars showed that single injections of HEDP or Cl2MDP induced subameloblastic cysts between the secretory ameloblasts and the developing enamel. The ameloblastic lining of the cysts contained numerous calcified deposits. A few days after injection, hypoplasias were seen in the enamel in areas previously occupied by cysts. In the erupted teeth, scanning electron microscopic examination revealed enamel hypoplasias which were mainly localized on the mesial sides of the cusps. In addition to the previously mentioned disturbances, multiple injections resulted in more extensive cysts, some of which contained non-mineralized enamel matrix. Inhibition of the mineralization of dentin and alveolar bone was also noticed.

In vitro differentiation of tooth rudiments of embryonic mice. II. Growth of isolated thirds of embryonic mouse incisors

AbstractThe mandibular incisor of the 16 day embryonic mouse was divided into three visibly equal thirds; each third was explanted in vitro to test its capabilities for independent development. Ameloblasts and odontoblasts, present in the distal third at the time of isolation, fail to continue their differentiation. This third flattens, and the tissues become disorganized. Grouping several distal thirds in a single culture did not promote tissue differentiation. The embryonic proximal third remains cohesive The tissues differentiate; both ameloblasts and odontoblasts deposit extracellular matrix. Approximately half of the cultures of the intermediate third show differentiated odontoblasts and extracellular matrix after 11 culture days. Measurements of the volume changes of ameloblasts, odontoblasts, and general mesenchyme were taken over the first six culture days for each third of the rudiment. The volume changes recorded, and the counts of mitotic figures reflect the developmental behavior of each fraction of the rudiment.

Ultrastructural changes in ameloblasts from fluoride treated rats

Repeated doses of fluoride over 2 days produced marked alterations in the fine structure of ameloblasts from tooth germs of young rats: large confluent distensions of the endoplasmic reticulum and swelling of isolated mitochondria. The appearance was similar to the documented material on initial degenerative changes in hepatocytes by a variety of other toxic agents. The possible role of these changes in the production of enamel fluorosis is discussed.