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[Histoautoradiographic studies on protein synthesis by odontoblasts].

The protein-producing function of the odontoblasts was studied in 120 Wistar rats with an initial weight ranging from 30 to 40 g determining the rate of the migration of radio-active isotopes (glycine-2-14C, lysine-1-14C) from the odontoblasts to the predentine. The odontoblasts exert a protein-synthetizing function and play an active part in predentine formation. Under the influence of a cariogenic saccharose-casein diet and the simultaneous induction of experimental caries, the intensity of the migration of the labelled amino acids from the odontoblasts to the predentine increases during the first and second stages of caries, which must be interpreted as an increase of the protein-synthetizing function of the odontoblasts associated with an intensification of dentinogenesis. During the third stage of caries, the protein-producing function of the odontoblasts decreases.

Animals↗

[Capdepont dysplasia and vertical excess of the lower half of the face. Genetic syndrome or fortuitous association?].

The study of an Algerian family revealed that four of the seven children presented dentinogenesis imperfecta associated with an anomaly of the facial skeleton (maxillary hypoplasia, anterior expansion of skeletal origin due to widening of the angle of the gonion). This type of association and its very unusual type of transmission (no doubt recessive) leads to a discussion of genetic mechanisms of these syndromes.

Child↗

[Histological data on the pharyngeal jaws of Trachinotus teraia (Cuvier 1832), Carangidae (Osteichthyes, Perciformes) of tropical Africa].

The thick pharyngeal jaws of Trachinotus teraia are made up of small teeth embedded in an abundant acellular vascularized bone tissue. The diameter of the vascular canals is progressively reduced from the center of the jaw to the abrasive surface where they are wholly obliterated by bone tissue. Each tooth consists of a small cone of orthodentine without an apical hypermineralized cape. The teeth have a weak masticatory function compared to the surrounding bone tissue. The genesis of these teeth probably takes place in the inner part of the pharyngeal jaws; they then move progressively towards the abrasive surface. The inner localization of the first stages of dentinogenesis raises the question of an effective epidermal participation in the ontogenesis of the pharyngeal teeth in T. teraia.

Animals↗

Dental findings in osteogenesis imperfecta: II. Dysplastic and other developmental defects.

Orthopantomograms of 49 patients with osteogenesis imperfecta (OI) classified according to Sillence were examined for dysplastic dentin defects and other developmental abnormalities of the teeth. Thistle-tube-shaped pulps in the permanent teeth were observed in five patients. Four of them had pulp stones in several teeth. Apically extended pulp chambers were present in 3/49 patients, and the structure of the mandible was cystic in 1/49. These defects occurred in patients with type I or unclassifiable OI and were mainly not associated with type I dentinogenesis imperfecta (DI). The prevalence rates of invaginations (10.2%) and hypodontia of permanent teeth (18.4%) exceeded those in the normal population. The frequent developmental disturbances of the teeth in OI may be secondary to the connective tissue defect. The relation of the dysplastic defects other than type I DI to OI remains to be clarified.

Connective Tissue↗

The time scale of tooth development and replacement in Xenopus laevis (Daudin).

One hundred and seventy two larval specimens of Xenopus laevis were reared in such a way that their rates of development (as measured by external criteria) were similar, and so the course of dental development could be examined histologically in a cross sectional study. In this way the events of tooth development were observed, and a time scale constructed for these events. The teeth took an average time of 26 days to develop, erupt and become ankylosed to the bony pedestal, after which each tooth was in a functional position for only about 7 days. Individual tooth replacement was assessed to occur about every 16 days. By comparing the number and size of the odontoblasts responsible for dentinogenesis in 18 months old adult Xenopus laevis with the odontoblasts in the larval specimens, the conclusion was drawn that, despite the larger size of the adult teeth, the time involved in their development and replacement may well be of similar duration to the smaller larval teeth. The significance of the findings for the Zahnreihen Theory is discussed.

Animals↗

A histological study of the innervation of developing mouse teeth.

The innervation of developing mouse teeth between initial formation and crown formation was investigated using silver-stained serial sections. The developing innervation correlated with the stage of development of individual teeth rather than the chronological age of the mice. Nerves approached the developing dental papilla during the bud stage and formed a basal plexus below the dental papilla in the early cap stage. Nerve fibres from this plexus spread into the dental follicle as it began to develop. However, nerves did not enter the dental papilla until crown formation commenced, when the innervation was fairly rapid. Innervation commenced in the incisor teeth as soon as dentinogenesis started but not until a thin layer of enamel had been formed in the molar teeth. Although some of the early fibres were associated with blood vessels, many nerves lay free in the pulp. The absence of nerves in intimate relationship to the presumptive dental regions during the inductive phase of tooth development suggests that neural induction plays no part in the initiation of odontogenesis. However, it is not possible, from a purely histological study such as this, to attribute any function to the nerves at other stages of tooth development until the neurotransmitter content, and hence the type and likely function of the nerves, is established.

Amelogenesis↗

From serum to the mineral phase. The role of the odontoblast in calcium transport and mineral formation.

Dentin may be considered as a calcified connective tissue and is in its composition as well as in its mode of formation closely related to bone. Dentin is formed by two simultaneous processes in which the odontoblasts are instrumental: the formation of the proteinaceous dentin matrix, and mineral crystal formation in this matrix. As part of this, the odontoblasts actively transport Ca2+ ions towards the site of mineral formation. The cells maintain a delicate intracellular Ca2+ ion balance by the concerted action of transmembraneous transport mechanisms, including Ca-ATPase, Na+/Ca2+ exchangers and calcium channels of the L-type, and possibly intracellular Ca(2+)-binding proteins. The net effect of this is a maintenance of a cytoplasmic sub-micromolar Ca2+ activity and an extracellular accumulation of Ca2+ ions at the mineralization front. In addition to the major matrix constituent, collagen, non-collagenous macromolecules, such as dentin phosphoprotein (phosphophoryn), dentin sialoprotein, and proteoglycan, are synthesized by the odontoblasts and deposited in the matrix. Such polyanionic macromolecules are presumably responsible for the extracellular induction of hydroxyapatite crystals, but may also function to inhibit mineral growth and to regulate crystal size. Accordingly, it can be concluded that dentinogenesis comprises an interplay between several factors in the tissue, cellular as well as extracellular.

Animals↗

[Dental complications of radiotherapy of tumors of the nasal cavity in childhood].

Authors have presented a case-report of a child with no or hardly developed roots of permanent teeth in the maxilla. Histological examinations of the removed teeth showed amelogenesis imperfecta and dentinogenesis imperfecta. The disturbance in root development is considered to be the consequence of the radiological treatment which was given to the child at the age of four because of the endodermal sinus tumor of the left middle nasal passage.

Amelogenesis Imperfecta↗

Dental manifestations of osteogenesis imperfecta and abnormalities of collagen I metabolism.

The in vitro protein-chemical features and the molecular background of osteogenesis imperfecta (OI), a heritable disorder of collagen I metabolism, have been elucidated in recent years. The aim of our study was to find the prevalence of dentinogenesis imperfecta (DI) and other dental anomalies in 88 patients with OI, to compare clinical with radiologic abnormalities, and to correlate these clinical/radiologic findings with the results of gel electrophoresis and molecular studies of collagen I. Twenty-eight percent of OI patients had DI. Most patients with DI had radiologic abnormalities, but some patients had radiologic signs compatible with DI, but no clinical signs of DI. OI type I patients with DI were more severely affected by OI than those without DI. In OI type III and IV, in contrast, there was no difference in overall severity between patients with and without DI. DI was not associated with any particular molecular aberration in any OI type. If defining DI from the presence of both clinical and radiologic signs, collagen I produced by cultured fibroblasts was qualitatively abnormal from all OI patients with DI. Some OI patients had dental abnormalities not resembling DI. A qualitative collagen abnormality could not be found in any of these patients. Denticles, i.e., calcifications within the pulpal cavity, were found more frequently in OI patients than in control subjects.

Adolescent↗

[Hereditary diseases with tooth anomalies and their causal genes].

In this review, we describe the current knowledge and the advances in research on human genes whose defect leads to dental anomalies. Recently, it was demonstrated that a missense mutation of a human homeobox MSX1 gene causes autosomal dominant agenesis of second premolars and third molars. X-linked anhidrotic ectodermal dysplasia (EDA), characterized by abnormal hair, teeth, and sweat glands, was demonstrated to be caused by a mutation in a novel transmembrane protein gene that is expressed in epithelial cells and in other adult and fetal tissues. The autosomal dominant Rieger syndrome (RS) manifests hypodontia, adontia, iridogoniodysgenesis and umbilical anomalies. Recently, a novel homeobox gene, RIEG, of Otx family was cloned as a causal gene of RS. The several mutations have been reported on the genes causing hypophosphatasia, which is characterized by defective mineralization of the skeletal and dental structures. An autosomal dominant dentinogenesis imperfecta (DI) is mostly associated with osteogenesis imperfecta (OI). Most patients with DI have mutations in either the COL1A1 or COL1A2 genes of type I collagen. Amelogenesis imperfecta (AI) is a diverse group of hereditary disorders characterized by a variety of developmental enamel defects including hypoplasia and hypomineralization, some of which have been revealed to be associated with defective amelogenin genes.

Alkaline Phosphatase↗

Osteogenesis imperfecta: mosaicism and refinement of the genotype-phenotype map in OI type III. Mutations in brief no. 242. Online.

Non-lethal OI III (OMIM 259420) is caused by structural aberrations of collagen I. We report four novel glycine substitutions, one in the a1 (I) chain of collagen I (G688S) and three in the a2 (I) chain (G241D, G247C, G883V). In each of two families (G241D and G883V), we found parental mosaicism for the substitution explaining recurrence and intrafamilial variability of OI. The G247C and the G883V are the most N-terminally and C-terminally, respectively, placed cysteine and valine substitutions reported. The new substitutions add important information to the genotype-phenotype map and in particular the importance of a-chain stoichiometry is underlined. Data regarding the G688S substitution may suggest a different effect of the two a-chains in the development of dentinogenesis imperfecta (DI).

Chromosome Mapping↗

Variation in the developing root cone angle of the permanent mandibular teeth of modern man and certain fossil hominids.

The radiographic appearance of the developing root apex of 461 mandibular teeth was studied using ellipsopantomographs of 77 individuals. The angle formed by the mesial and distal aspects of the developing root cone was measured for each quarter of root development in the permanent mandibular teeth of each subject. Large values for this angle in the more coronal part of the root gradually decreased towards the root apex where values were small in all teeth studied. This gradual decrease in the angle of the developing root cone is concordant with the faster increase in root length but slower rates of dentinogenesis reported for the apical region of human teeth in the literature, as the decrease is likely to reflect the increasing numbers of odontoblasts that become involved in root formation as it proceeds apically. The developing root cone angles of one of the five fossil hominid specimens studied (KMN-ER 820) fall outside the range for the developing root cone angle of the developing premolar and permanent second molar roots in modern Homo sapiens and is evidence of a faster rate of root elongation than is typical for modern man.

Animals↗

The effect of beta-aminoproprionitrile on the periodontal ligament: II. Radioautographic study of collagen secretion from fibroblasts.

Fibroblasts are distributed evenly throughout the periodontal ligament (PDL) of normal mice. In mice fed beta-aminoproprionitrile (beta-APN) the fibroblasts undergo aggregation to form palisades of closely juxtaposed cells abutting pools of acellular collagenous matrix. Individual fibroblasts within these aggregates retain their polarized cytoplasmic organization and continue to synthesize and secrete collagen. However, unlike normal PDL fibroblasts, the beta-APN-treated cells appear immobilized by well-developed cell-to-cell adherens-type junctions along their lateral surfaces. We studied collagen secretion from beta-APN-treated fibroblasts by light and electron microscopic radioautography after injection of 3H-proline. Newly synthesized collagen was secreted from the distal ends of the beta-APN-aggregated fibroblasts as a distinct band of labeled material, resembling the pattern of matrix deposition seen in osteogenesis and dentinogenesis. The radioactive band of collagenous matrix was displaced further away from the fibroblasts at 2 and 4 days after 3H-proline injection as more collagen was secreted. This pattern of radiolabeled collagen secretion confirmed previous observations that PDL fibroblasts are highly polarized and that collagen secretory granules are extruded from the distal or secretory pole of the cell. In normal PDL the even distribution of fibroblasts and the complex interrelationship of their distal cell processes leads to a diffuse pattern of silver grain deposition, masking the oriented flow of new collagen from the distal ends of individual fibroblasts. Analysis of electron microscopic radioautographs revealed that newly synthesized collagen was packaged and secreted from beta-APN-treated fibroblasts via the normal cytoplasmic pathways but at a slower rate.

Aminopropionitrile↗

Odontoblast-specific expression of cre recombinase successfully deletes gene segments flanked by loxP sites in mouse teeth.

Embryonic or neonatal lethality of mice with targeted disruption of critical genes preclude them from further characterization of specific roles of these genes during postnatal development and aging. In order to study the molecular roles of such genes in teeth, we generated transgenic mouse lines expressing bacteriophage Cre recombinase under the control of the mouse dentin sialophosphoprotein (dspp) gene promoter. The expression of Cre recombinase protein was mainly detected in the nucleus of the odontoblasts. The efficiency of Cre activity was analyzed by crossing the Dspp-Cre mice with ROSA26 reporter (R26R) mice. The offspring with both genotypes have shown specific deletion of intervening sequences flanked by loxP sites upstream of the reporter gene, thereby facilitating the expression of the beta-galactosidase (beta-gal) gene in the teeth. The activity of beta-gal was initially observed in the odontoblasts of 1-day-old mice and increased with tooth development. Almost all of the odontoblasts have shown lacZ activity by 3 weeks of age. We could not detect Cre recombinase activity in any other cells, including ameloblasts. These studies indicate that the Dspp-Cre transgenic mice will be valuable to generate odontoblast-specific gene knockout mice so as to gain insight into the molecular roles of critical genes in the odontoblasts during dentinogenesis.

Animals↗

Recurrence of lethal osteogenesis imperfecta due to parental mosaicism for a mutation in the COL1A2 gene of type I collagen. The mosaic parent exhibits phenotypic features of a mild form of the disease.

We have determined that a man, ascertained because he fathered a child with lethal osteogenesis imperfecta (OI) with each of two partners, is mosaic in both his germline and somatic tissues for a mutation in the COL1A2 gene which encodes the pro alpha 2(I) chain of type I procollagen. His dermal fibroblasts were previously shown to synthesize a population of cysteine-containing alpha 2(I) chains that were posttranslationally overmodified. DNA sequence analysis of COL1A2 cDNAs demonstrated that the cysteine-containing chain resulted from a point mutation (G to T) in the first position of the codon for the glycine at residue 472 of the triple helical domain. Genomic DNA from the one available affected infant contained the mutant and normal COL1A2 alleles in equal proportion. Examination of DNA from several tissues of the father showed that the mutant allele was present in approximately 40% of his sperm, 80% of his lymphocytes, and nearly 100% of his dermal fibroblasts. Despite the high level of mosaicism detected in somatic tissues, the only phenotypic manifestation of OI in the proband was that he was shorter than his unaffected male relatives and had mild dentinogenesis imperfecta. Thermal stability of type I collagen molecules containing the substitution was decreased, but to a lesser extent than for a nonlethal cysteine for glycine substitution at residue 259 of alpha 2(I), indicating that this measure of molecular stability may be of limited use in explaining the pathogenesis of osteogenesis imperfecta.

Adult↗

Development expression of bone sialoprotein mRNA in rat mineralized connective tissues.

Bone sialoprotein (BSP) is a phosphorylated and sulfated glycoprotein that is a major noncollagenous protein of bone and other mineralizing connective tissues. BSP is characterized by the presence of several polyglutamic acid segments and an RGD motif that mediates cell attachment through a vitronectin-like receptor. Although the precise function of BSP is unknown, the expression of BSP in conjunction with bone formation in vitro indicates a role for this protein in the biomineralization of connective tissues. In this study we used Northern hybridization and in situ hybridization to determine the tissue-specific and developmental expression of BSP during embryogenesis and growth of rat tissues. Analysis of tissues obtained from 13, 17, and 21 day fetuses, and from 4-, 14-, and 100-day-old animals indicates that BSP mRNA expression is restricted to cells actively forming the mineralizing tissues of bone, dentin and cementum. BSP mRNA transcripts were first evident in fully differentiated osteoblasts of 17 day fetal tissues at sites of de novo intramembranous and endochondral bone formation, with maximal expression observed at 21 days of gestation. Thereafter, BSP mRNA levels decreased markedly, and in adult bone hybridization was detected only in the primary spongiosa of long bones. In comparison, mRNAs for osteopontin (OPN), alkaline phosphatase (ALP), and osteocalcin (OC) peaked at 4-14 days postpartum before declining. In the tibiae, Northern hybridization revealed a second peak of mRNA for BSP, ALP, and OPN at 14 days, reflecting an increased osteogenic activity due to the formation of the secondary centers of ossification in the epiphyseal cartilage. In situ hybridization also revealed BSP mRNA in hypertrophic chondrocytes at sites of bone formation, in odontoblasts of the incisor during dentinogenesis, and in cementoblasts during cementogenesis. In view of the restricted distribution and temporal changes in the expression of BSP mRNA that we observed together with the chemical properties of BSP, we believe that this protein has a specific role in mediating the initial stages of connective tissue mineralization.

Alkaline Phosphatase↗

Immunolocalization of osteopontin, osteocalcin, and dentin sialoprotein during dental root formation and early cementogenesis in the rat.

Using immunohistochemical methods we studied the tissue localization of the extracellular matrix proteins osteopontin (OPN), osteocalcin (OC), and dentin sialoprotein (DSP) during the formation of acellular and cellular cementum in newly born rats. In the layer of acellular cementum of developing incisor and molar teeth we found a very strong staining for OPN but not for DSP or OC. Many cells immediately adjacent to acellular cementum and PDL cells were also positive for OPN but not for DSP or for OC. In contrast, cellular cementum in molar teeth stained strongly for OPN and OC but not for DSP. Consistent with these observations, the cells engaged in the formation of cellular cementum (cementoblasts and cementocytes) reacted strongly for OPN and OC but not for DSP. In advanced stages of dentinogenesis, both crown and root odontoblasts and dentin stained for OPN, OC, and DSP. Cells and matrices of surrounding alveolar bone stained for OPN and OC but not for DSP. We conclude that cementoblasts and cementocytes of cellular cementum produce OPN and OC but not DSP and thus express an osteoblast-like, not an odontoblast-like, phenotype. The cells responsible for the production of acellular cementum are likely cells of the PDL in close contact with the dental root surface. These fibroblast-like cells express OPN but not OC or DSP and accordingly express only a partial osteoblastic phenotype.

Animals↗

Structural features of forming and developing blood capillaries of the enamel organ of rat molar tooth germs observed by light and electron microscopy.

The process of vascularization of the enamel organ, a unique epithelial structure, occurs when the tooth germ is fully developed, i.e., at the onset of dentinogenesis. Although the three-dimensional organization of the capillaries has been previously investigated, the structural features underlying the formation of the new capillaries remains poorly understood. Thus, in the hope of better understanding the mechanism of formation of the stellate reticulum capillaries, upper first molar tooth germs of newborn and 3-day-old rats were fixed in glutaraldehyde-formaldehyde and processed for light and electron microscopy. Our results showed that blood capillaries are initially in close proximity to the outer enamel epithelium. Between and intercalated with the capillaries are round/ovoid clusters of cells, some of which are vacuolated, closely apposed to the outer enamel epithelium. The outer enamel epithelium is not a continuous layer, but exhibits gaps between the cells. This suggests that the capillaries penetrate the enamel organ through these gaps, since no invagination of the epithelium was observed. The presence of a cluster of cells containing vacuoles suggests that vasculogenesis is taking place. Images showing loss of the basal lamina, proliferation of endothelial cells, presence of filopodia and lateral sprouting suggests that angiogenesis is also occurring. Thus, neoformation of capillaries of the molar enamel organ of rat seems to occur simultaneously by mechanisms of vasculogenesis and angiogenesis.

Animals↗