Birefringence of the secretory-stage enamel organic extracellular matrix from rats submitted to successive injections of bisphosphonates.

The aim of the present study was to assess birefringence of the secretory-stage enamel organic extracellular matrix (ECM) and mechanical properties of mature enamel from rats treated with bisphosphonates. Longitudinal sections were obtained from upper incisors of rats that had been submitted to injections of bisodic etidronate (8 mg/Kg/day), sodium alendronate (30 microg/Kg/day), or sodium chloride as control (8 mg/Kg/day) for 42 days. Sections were immersed in 80% glycerin for 30 min and optical retardation of birefringence brightness in the secretory-stage enamel organic ECM was determined in nanometers. Etidronate-treated rats exhibited extensive morphological changes in the secretory-stage enamel organic ECM inclusive nonbirefringent conspicuous incremental lines, but presented optical retardation values similar to those showed by control rats (p > 0.05). Birefringence of secretory enamel organic ECM from etidronate rats presented an irregular aspect. Alendronate-treated rats did not show morphological alterations in the secretory-stage enamel organic ECM, however, they presented significant reduction in optical retardation of birefringence brightness when compared with control and etidronate rats (p < 0.01). Alendronate and etidronate groups exhibited reductions of approximately 6-10% in mature enamel cross-sectional microhardness when compared with control group (p < 0.01). Scanning electron microscopy analysis showed extensive alterations in mature enamel only from etidronate group with absence of enamel rods. The present work shows that bisphosphonates can affect the birefringence of the secretory-stage enamel organic ECM. The results presented here suggest that alterations in the supramolecular organization of the secretory-stage enamel organic ECM are a plausible mechanism by which environmental factors may cause enamel defects.

Anisotropic properties of the enamel organic extracellular matrix.

Enamel biosynthesis is initiated by the secretion, processing, and self-assembly of a complex mixture of proteins. This supramolecular ensemble controls the nucleation of the crystalline mineral phase. The detection of anisotropic properties by polarizing microscopy has been extensively used to detect macromolecular organizations in ordinary histological sections. The aim of this work was to study the birefringence of enamel organic matrix during the development of rat molar and incisor teeth. Incisor and molar teeth of rats were fixed in 2% paraformaldehyde/0.5% glutaraldehyde in 0.2 M phosphate-buffered saline (PBS), pH 7.2, and decalcified in 5% nitric acid/4% formaldehyde. After paraffin embedding, 5-microm-thick sections were obtained, treated with xylene, and hydrated. Form birefringence curves were obtained after measuring optical retardations in imbibing media, with different refractive indices. Our observations showed that enamel organic matrix of rat incisor and molar teeth is strongly birefringent, presenting an ordered supramolecular structure. The birefringence starts during the early secretion phase and disappears at the maturation phase. The analysis of enamel organic matrix birefringence may be used to detect the effects of genetic and environmental factors on the supramolecular orientation of enamel matrix and their effects on the structure of mature enamel.

Development of functional dentin incisors after a partial resection of the odontogenic organ of rat incisors.

The resection of the labial half of the odontogenic organ of rat incisors resulted in the development of teeth without enamel. Ten out of 26 operated rats developed a functional dentin incisor, i.e. a continuously growing and erupting tooth. These teeth were a little shorter and much thinner than normal incisors. The dentin and pulp presented a normal structure. Periodontal ligament and cement started to develop at the lingual face and gradually all tooth faces were invested by these tissues. The original socket space, to accommodate a thinner tooth, was narrowed by newly formed bone around the inner face of the socket. Eleven rats developed defective dentin incisors; these teeth showed signs of growth, however, their eruption was impaired. The operation failed in five rats. The odontogenic organ of the dentin incisor presented islands of epithelial cells at the labial aspect of a dense mass of mesenchyme cells. These islands, formed by densely packed, dark-staining cells encircling a few pale-staining cells, merged gradually, forming a root sheath and a cervical loop limiting a long apical foramen. The bulk of the bulbous part (apical bud) was absent; thus, there was no differentiation of ameloblasts and of the crown-analogue part of the incisor. The growth and eruptive behaviour of the dentin incisor, similar to that of a normal incisor, indicates that it has to bear a stem cell niche to retain its regenerative capacity. As in the apical bud, this niche is apparently located at the stellate reticulum of the cervical loop. The putative molecular mechanisms related to either the maintenance of the stem cell niche or the differentiation of the enamel organ and the root sheath are discussed. These data and our results, showing the development of a functional dentin incisor, suggest that the root-analogue part of the rodent incisor is an anatomic-physiological entity.

Ultramorphological analysis of resin-dentin interfaces produced with water-based single-step and two-step adhesives: nanoleakage expression.

This study evaluated the nanoleakage patterns in bonded interfaces using two single-step, self-etching adhesives (Adper Prompt-AD, and One-up Bond F-OB), two two-step, self-etching primers (Clearfil SE Bond-CF, and Unifil Bond-UB), and one two-step, total-etch adhesive (Single Bond-SB). Dentin surfaces were bonded with the adhesive systems and stored in water at 37 degrees C for 1 week and 6 months. After storage periods, teeth were sectioned into 0.8 mm-thick slabs, coated with nail varnish except for the bonded interfaces, and immersed in ammoniacal AgNO(3) for 24 h. After immersion in photodeveloping solution, bonded sections were prepared and observed under a SEM using the backscattered electron mode. Undemineralized, unstained, epoxy resin-embedded sections were prepared for TEM. Nanoleakage patterns were qualitatively compared between periods. Nanoleakage was observed in all bonded specimens at both periods. CF and UB presented silver deposits predominantly restricted to the thin (0.5 microm) hybrid layer (HL) at both periods. Although no evident differences were observed in the nanoleakage pattern of UB at 7 days and 6 months, CF presented enlarged areas of silver impregnation after 6 months. SB presented accumulation of silver particles mostly within the HL at 7 days, which was intensified after 6 months. AD and OB presented massive silver accumulation within the HL and the overlying adhesive layer. No evident differences were noticed between storage periods. Silver impregnation increased for all adhesive systems from 7 days to 6 months, except for UB.

The effect of partial damage to the enamel-related periodontium combined with root resection on eruption of the rat incisor eruption.

Previous work has indicated that the enamel-related periodontium (ERP) has a role in the eruptive process of the rat lower incisor. By combining partial damage of this tissue with resection of the odontogenic organ, we examined the effect of the damage on subsequent incisor eruption. The connective tissue of the enamel-related periodontium was regenerated in less than 2 weeks, showing morphology close to normal. The injured part of the enamel organ was neither regenerated nor repaired, and a cement-like tissue, continuous with the true acellular cement, was formed on the denuded enamel. Before tooth exfoliation, the operated teeth erupted at a slower rate compared with root-resected and sham-operated incisors, probably because of the absence of a substantial part of the enamel organ due to surgical damage. As with the coronal dental follicle and the enamel organ in rat molars, the enamel-related periodontium and the enamel organ of rat incisors may have some control on their eruptive process.

An ultrastructural study of the supporting tissues of sea urchin (Lytechinus variegatus) teeth

The teeth of sea urchins are connected to the calcareous jaw plates, known as pyramids, by a ligament consisting of collagen fibers and microfibrils synthesized by fibroblasts in the aboral growth zone of the tooth. This ligament needs to be sufficiently stiff to hold the teeth firmly when the animal scrapes hard surfaces, but also needs to be flexible enough to allow the teeth to move outwards during growth. To understand the mechanisms that regulate the growth and stiffness of sea urchin teeth, we have examined the ultrastructural organization of the supporting structures of Lytechinus variegatus teeth. Electron microscopy showed that collagen fibrils were mechanically attached to the jaws was formed by cavities that ramificated in the deep portions. The collagen fibrils were not mechanically linked to the jaws. These findings suggest that the stiffness of the ligament is mediated by chemical bonding between the collagen fibrils and the jaw surface. The cavities present in the pyramids greatly increased the surface area and strengthened the area for the bonding of collagen fibrils.