Proteomic Mapping of Dental Enamel Matrix from Inbred Mouse Strains: Unraveling Potential New Players in Enamel.

Enamel formation is a complex 2-step process by which proteins are secreted to form an extracellular matrix, followed by massive protein degradation and subsequent mineralization. Excessive systemic exposure to fluoride can disrupt this process and lead to a condition known as dental fluorosis. The genetic background influences the responses of mineralized tissues to fluoride, such as dental fluorosis, observed in A/J and 129P3/J mice. The aim of the present study was to map the protein profile of enamel matrix from A/J and 129P3/J strains. Enamel matrix samples were obtained from A/J and 129P3/J mice and analyzed by 2-dimensional electrophoresis and liquid chromatography coupled with mass spectrometry. A total of 120 proteins were identified, and 7 of them were classified as putative uncharacterized proteins and analyzed in silico for structural and functional characterization. An interesting finding was the possibility of the uncharacterized sequence Q8BIS2 being an enzyme involved in the degradation of matrix proteins. Thus, the results provide a comprehensive view of the structure and function for putative uncharacterized proteins found in the enamel matrix that could help to elucidate the mechanisms involved in enamel biomineralization and genetic susceptibility to dental fluorosis.

In vitro re-hardening of artificial enamel caries lesions using enamel matrix proteins or self-assembling peptides.

OBJECTIVES: To assess the re-hardening potential of enamel matrix derivatives (EMD) and self-assembling peptides in vitro, hypothesizing that these materials may increase the mineralization of artificial carious lesions and improve hardness profiles. MATERIAL AND METHODS: Forty-eight enamel samples were prepared from extracted bovine lower central incisors. After embedding and polishing, nail varnish was applied, leaving a defined test area. One third of this area was covered with a flowable composite (non-demineralized control). The remaining area was demineralized in an acidic buffer solution for 18 d to simulate a carious lesion. Half the demineralized area was then covered with composite (demineralized control), while the last third was left open for three test and one control treatments: (A) Application of enamel-matrix proteins (EMD - lyophilized protein fractions dissolved in acetic acid, Straumann), (B) self-assembling peptides (SAP, Curodont), or (C) amine fluoride solution (Am-F, GABA) for 5 min each. Untreated samples (D) served as control. After treatment, samples were immersed in artificial saliva for four weeks (remineralization phase) and microhardness (Knoop) depth profiles (25-300 µm) were obtained at sections. Two-way ANOVA was calculated to determine differences between the areas (re-hardening or softening). RESULTS: Decalcification resulted in significant softening of the subsurface enamel in all groups (A-D). A significant re-hardening up to 125 µm was observed in the EMD and SAP groups. CONCLUSIONS: This study showed that EMD and SAP were able to improve the hardness profiles when applied to deep demineralized artificial lesions. However, further research is needed to verify and improve this observed effect.

In vitro re-hardening of artificial enamel caries lesions using enamel matrix proteins or self-assembling peptides

ABSTRACT Objectives To assess the re-hardening potential of enamel matrix derivatives (EMD) and self-assembling peptides in vitro, hypothesizing that these materials may increase the mineralization of artificial carious lesions and improve hardness profiles. Material and Methods Forty-eight enamel samples were prepared from extracted bovine lower central incisors. After embedding and polishing, nail varnish was applied, leaving a defined test area. One third of this area was covered with a flowable composite (non-demineralized control). The remaining area was demineralized in an acidic buffer solution for 18 d to simulate a carious lesion. Half the demineralized area was then covered with composite (demineralized control), while the last third was left open for three test and one control treatments: (A) Application of enamel-matrix proteins (EMD - lyophilized protein fractions dissolved in acetic acid, Straumann), (B) self-assembling peptides (SAP, Curodont), or (C) amine fluoride solution (Am-F, GABA) for 5 min each. Untreated samples (D) served as control. After treatment, samples were immersed in artificial saliva for four weeks (remineralization phase) and microhardness (Knoop) depth profiles (25-300 µm) were obtained at sections. Two-way ANOVA was calculated to determine differences between the areas (re-hardening or softening). Results Decalcification resulted in significant softening of the subsurface enamel in all groups (A-D). A significant re-hardening up to 125 µm was observed in the EMD and SAP groups. Conclusions This study showed that EMD and SAP were able to improve the hardness profiles when applied to deep demineralized artificial lesions. However, further research is needed to verify and improve this observed effect.

Amelogenin and enamelysin localization in human dental germs.

Odontogenesis is extensively studied in animal models but less understood in human. In early amelogenesis, amelogenin constitutes 90% of enamel organic matrix, which is degraded by enamelysin and replaced by hydroxyapatite crystals. Here, amelogenin and enamelysin distribution changes during amelogenesis were shown by co-localization experiments by confocal microscopy. Early bell stage showed more amelogenin labeling than enamelysin, as free immune-reactive granular patches towards basal membrane between ameloblast and odontoblast. Increased amelogenin expression and secretion towards extracellular matrix formation region was found. Enamelysin distribution was perinuclear in early bell stage. During late bell stage, a decreasing amelogenin labeling in contrast with enamelysin increasing along the cells was found, suggesting specific temporal amelogenin degradation. Enamelysin was located initially around nuclei and later was found in all the ameloblast and stellate reticulum cytoplasm. Amelogenin was observed inside ameloblast, stellate reticulum, and intermediate stratum cells in the enamel as well as in the newly formed dentin extracellular matrix. In contrast, in dentin more amelogenin than enamelysin was found located close to the periphery.

Resin replica in enamel deproteinization and its effect on acid etching.

PURPOSE: The goal of this in vitro study was to identify the topographical features of deproteinized (NaOCl) and etched with phosphoric acid (H3PO4) enamel surface, compared to phosphoric acid surface alone with a Resin Replica model. MATERIALS: Ten extracted lower first and second permanent molars were polished with pumice and water, and then divided into 3 equal buccal sections having similar physical and chemical properties. The enamel surfaces of each group were subjected to the following treatments: Group A: Acid Etching with H3PO4 37% for 15 seconds. Group B: Sodium Hypochlorite (NaOCl) 5.25% for 60 seconds followed by Acid Etching with H3PO4 37% for 15 seconds. Group C; No treatment (control). All the samples were treated as follow: Adhesive and resin were applied to all groups after A, B and C treatment were performed; Then enamel/dentin decalcification and deproteinization and topographic SEM Resin Replica assessment were used to identify resin tags enamel surface quality penetration. RESULTS: Showed that group B reached an area of 7.52 mm of the total surface, with a 5.68 mm2 (73%) resin tag penetration equivalent type I and II etching pattern, 1.71 mm2 (26%) equivalent to type III etching pattern and 0.07 mm2 (1%) unaffected surface. Followed by group A with 7.48 mm2 of the total surface, with a 3.47 mm2 (46 %)resin tag penetration equivalent to type I and II etching pattern, 3.30 mm2 (45%)equivalent to type III etching pattern and 0.71 mm2, and (9%) unaffected surface. Group C did not show any resin tag penetration. A significant statistical diference (P < 0.001) existed between groups A and B in resin quality penetration, leading to the conclusion that when the enamel is deproteinizated with 5.25% NaOCl for 1 minute prior H3PO4, the surface and topographical features of the replica resin penetration surface increases significantly with type I-II etching pattern.