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PubMed1993

Relationships between mandibular canine calcification stages and skeletal maturity.

Hand wrist radiographs and dental panoramic radiographs of 200 boys and 215 girls were assessed to determine the relationship between the developmental stages of the mandibular canine and skeletal maturity indicators of the pubertal growth spurt. Chi-square and Kendall's tau statistics demonstrated significant (P < 0.001) relationships between skeletal maturity indicators. Associations between canine development and skeletal maturity ranged from 0.53 to 0.85. Most children having attained canine stage G showed the presence of the adductor sesamoid (81%), capping of the diaphysis of the third middle phalanx (77%), and capping of the fifth proximal phalanx (87%). Growth reference data for American children suggest that stage G occurred approximately 0.4 years and 1.3 years before peak height velocity for girls and boys, respectively. It is indicated that canine calcification could serve as a useful tool for evaluating childrens' skeletal maturation and, by association, somatic maturity.

Adolescent↗

PubMed2004

Accelerated bone formation and increased osteoblast number contribute to the abnormal tooth germ development in parathyroid hormone-related protein knockout mice.

Our previous study showed that tooth germs at late embryonic stage [later than embryonic day 17.5 (E17.5)] and neonatal homozygous parathyroid hormone-related protein (PTHrP)-knockout mice are compressed or penetrated by the surrounding alveolar bone tissue. In vivo and in vitro studies have shown that the development of the tooth germ proper is not disturbed, but insufficient alveolar bone resorption, due to the decreased number and hypofunction of osteoclasts, is the main cause of this abnormality. In addition to the insufficient alveolar bone resorption, progressive bone formation toward tooth germs was observed in homozygous mice, suggesting that accelerated bone formation also contributes to this abnormality. To further investigate this, homozygous mice at E14.0 and E15.5, when alveolar bone is forming, were used for histochemical and bone histomorphometric analyses. In contrast to the late embryonic stage, the alveolar bone did not yet compress developing tooth germs in homozygous mice on E14.0, but a larger amount of bone tissue was seen compared to wild-type littermates. Histomorphometric analysis of bone at E14.0 revealed that the osteoblast numbers and surfaces in the mandibles and in the bone collar of femora of homozygous mice were significantly higher than those of wild-type mice. However, unlike our previous study showing the osteoclast surface on E18.5 in homozygous mice to be significantly lower than that of wild-type mice, this study at E14.0 showed no significant difference between the two genotypes. To evaluate the amount of calcification around tooth germs, 3D images of mandibles were reconstructed from the calcein-labeled sections of the wild-type and mutant mice. Labeling was performed at E14.0, and the mice were sacrificed 1 h after the calcein injection to minimize the effect of bone resorption. Comparison of the 3D images revealed that the labeled surface was larger around developing tooth germs in homozygous mouse than in wild-type mouse. On day E15.5, osteoblasts approached the enamel organ of homozygous mice but this was not observed in wild-type mice. In this study, we report a systemic increase in osteoblast number and accelerated bone formation in homozygous PTHrP-knockout mice, both of which contribute to the abnormal tooth development.

Acid Phosphatase↗

PubMed1994

Morphological studies of hypomineralized enamel of rat pups on calcium-deficient diet, and of its changes after return to normal diet.

BACKGROUND: Micro-hardness investigations have shown that rat pups nursed by mothers on a low calcium diet and weaned with the maternal calcium-deficient diet develop hypomineralized enamel. The inorganic and organic components of this enamel, their relationships, and their changes after return to normal diet have been studied by light and electron microscopy. METHODS: The maturation zone of incisor enamel has been studied in: (1) rats nursed for 20 days by mothers on a low calcium diet and weaned for 30 days with the same diet (E1 enamel); (2) rats that after the calcium-deficient diet were fed normal diet for 10 days (E2 enamel); and (3) rats nursed for 20 days by mothers on a normal diet and weaned for 30 days with a normal diet (controls). RESULTS: The results showed that E1 enamel was hypomineralized, as noted by its Azure II-Methylene blue stainability in undecalcified sections, its light staining with the von Kossa method, and its ultrastructure. E1 crystallites, although present throughout the whole enamel, were thinner than those of E2 enamel, which were similar to those of controls. E1 interrod crystallites were thicker in the intermediate than in the dentinal zone and were thicker than rod crystallites. Organic matrix was present throughout the whole E1 enamel. Its organic components (crystal ghosts) had the same shape, arrangement, and organization as those of inorganic crystallites. Crystal ghosts were greatly reduced in E2 enamel and in controls. CONCLUSIONS: The results lead to the conclusions that: (1) E1 enamel is hypomineralized, and its degree of calcification is restored by return to a normal calcium diet; (2) intra- and interprismatic calcification occurs in a different way; (3) crystallite thickness is initially greater in dentinal than in the superficial zone and is reversed as crystallite growth is completed; and (4) loss of enamel proteins is necessary for completion of crystallite growth and not for crystallite formation.

Animal Nutritional Physiological Phenomena↗

PubMed2002

[The study of calcification of autogenous bone marrow stem cell transplantation on alveolar bone defect in dogs].

OBJECTIVE: To evaluate the effect of calcification of autogenous bone marrow stem cell transplantation in periodontal tissue regeneration. METHODS: Bone marrow stem cells derived from the same dog were cultured with alpha-MEM. 1 x 10(7) cells of first passage were allowed to attach to the collagen membrane for 24 hours. The membrane-cells were transplanted into periodontal defect in the same dog. Then the defects were covered with e-pTFE membranes. The defects covered only with e-pTFE without membrane-cells were served as control. Eighteen teeth of 6 dogs for every group were studied. The dogs were sacrificed after 6 weeks. RESULTS: The results showed that new bone formation in test group was significantly higher than that of control group. The calcification of new bone in test group was better than control group. CONCLUSIONS: The results suggested that autogenous bone marrow stem cell transplantation with guided tissue regeneration technique could enhance periodontal tissue regeneration and could form new bone tissue fast and could shorten times of periodontal tissue regeneration in dogs.

Alveolar Bone Loss↗

PubMed2001

The in vitro and in vivo influence of 4-META/MMA-TBB resin components on dental pulp tissues.

The purpose of this study was to qualitate the penetration of the major components of 4-META/MMA-TBB adhesive resin (4-META resin) and to characterize their influence on the in vitro and in vivo wound healing of dental pulp tissues. Fresh 4-META resin was applied to rabbit mesentery; its components penetrated the mesentery to form three of layers, depending on the amounts of monomer components in the tissue. The superficial layer was a soft-tissue hybrid layer (STHL), the intermediate layer contained small particles of polymerized 4-META resin, while the deepest layer contained unpolymerized monomer components including MMA and butanol, which were detected by gas chromatography (GC). To characterize the in vivo effects of the deepest layer, we immersed the pulp tissue in MMA or in 5% 4-META/MMA and autotransplanted it to placement beneath a rabbit kidney capsule. The MMA-immersed pulp was positive for osteocalcin and presented osteodentin formation at 7 days, as did the untreated control pulp tissue. In contrast, the 5% 4-META/MMA-immersed pulp collapsed into the cell-deficient fibrous connective tissue, with slight calcification by 7 days and less osteodentin formation at 14 days. Analysis of these data suggests that MMA does not inhibit osteogenic activity of pulp tissue, while 5% 4-META/MMA does inhibit osteogenic activity to some extent.

Acrylic Resins↗

PubMed1982

Primary mineralization of dentin in rats after pulp capping with calcium-hydroxide.

It has been previously reported that short term application of calcium-hydroxide to the dental pulp resulted in the formation of a dentin bridge with matrix vesicle calcification. The present study deals with the induction of reparative dentin in rat molar pulps by direct capping with calcium-hydroxide. The teeth were examined after 21 and 35 days. Transmission electron microscopy (TEM) revealed that the calcification process in the newly formed dentin was characterized by apatite deposition in matrix vesicles and the occurrence of calcifying nodules in the matrix. The persistence of these features of primary mineralization, during the experimental period, was associated with the continuous calcium hydroxide contact with pulp tissue, producing changes in its metabolic state.

Animals↗

PubMed2000

Approaches to chronological age assessment based on dental calcification.

The aim of the present study was to find an accurate estimation of chronological age using a small number of selected teeth. For this purpose, the method devised by Nolla [C. Nolla, The development of the permanent teeth, J. Dent. Child. 27 (1960) 254-266.] was used: the development of each of the teeth was determined according to this method on 374 radiographs, 195 of boys (mean age 8.59) and 179 of girls (mean age 8.75). The 28 variables representing the calcification stages were analyzed using cluster analysis followed by multivariate analysis (multiple linear regression model). Patient age was considered to be a dependent variable. Our study showed that antimere teeth are the most homogeneous as regards stages of development. The prediction was more accurate for boys and girls below 10 years of age, using teeth 21, 43 and 46 from boys and teeth 21, 46 and 47 from girls. These teeth accounted for 80% total variance of chronological age for dental calcification. Standard error was +/-1.4 years for boys and +/-1.2 years for girls. When the age of the children remained completely unknown, the best estimates were provided by teeth 43, 47, 46 and 44 from boys and teeth 44, 47 and 43 from girls.

Adolescent↗

PubMed1989

X-ray crystallographic studies as to the calcification in the Hunter-Schreger bands of human enamel.

By analyzing integrated intensities of X-ray diffraction patterns and X-ray diffractograms obtained from the two zones of the Hunter-Schreger bands, it was deduced that the ratio of the sums of the integrated intensities and the degree of preferred orientation of the crystal aggregates in both zones of the bands were almost the same in value, but the fiber axes of the two zones were oriented in different directions. From these experimental observations and a consideration of the effect of the orientation of the enamel prisms relative to the incident X-ray beam, it is concluded that the two zones of the bands were calcified evenly and the light and dark zones on the microradiographs were caused by the difference in the orientation of enamel prisms.

Bicuspid↗

PubMed2006

Effects of pH on mineralization ability of human dental pulp cells.

The purpose of this study was to investigate the effect of alkaline pH on calcification in human dental pulp (HDP) cells. HDP cells were cultured in pH 7.8 conditioned medium, and alkaline phosphatase (ALP) activity was measured. The ALP activity was higher in the pH 7.8 conditioned medium group than in the pH 7.2 conditioned medium group. Expression of mRNAs for bone morphogenetic protein (BMP)-2 was measured by the RT-PCR technique. The expression of BMP-2 in the pH 7.8 groups was greater than that in the pH 7.2 group. Furthermore, we determined Calcified nodule formation by von Kossa staining. The number of calcified nodules was increased in the pH 7.8 conditioned medium. These results suggest that HDP cell mineralization was enhanced in alkaline pH (pH 7.8) conditioned medium.

Alkaline Phosphatase↗

PubMed1989

Dentin matrix proteins: composition and possible functions in calcification.

Dentin may be regarded as a mineralized connective tissue. In its composition as well as its mode of formation, dentin exhibits several similarities with bone, but also definite differences. The dentin organic phase, the matrix, determines its morphology and is believed to be instrumental in the formation of the mineral phase. A fibrous web of collagen type I dominates the organic matrix. Also, minor amounts of other collagen types may be present. The noncollagenous proteins (NCPs), which constitute about 10% of the matrix, fall into several categories: phosphoproteins, Gla-proteins of the osteocalcin type as well as matrix Gla-protein, proteoglycans, different acidic glycoproteins, and serum proteins. Some of these NCPs have unique chemical compositions that give them specific properties. Dentinogenesis occurs by two simultaneous processes: the formation of a collagenous web in predentin, which is followed by the formation of the inorganic phase at the mineralization front. The composition of the predentin organic matrix differs from that of dentin, as some NCP components are secreted extracellularly just in advance of the mineralization front. In addition, some constituents of predentin seem to be metabolized. The NCPs may be important to several processes during dentinogenesis. Much evidence indicates that noncollagenous components in the matrix are instrumental in mineral formation. New data show that polyanionic NCPs, such as phosphoprotein and proteoglycans, when immobilized on a solid support, induce apatite formation under physiological conditions. These data indicate that polyanionic NCPs may function as mineral nucleators in vivo. They may also act as size and rate regulators for crystallization and promote calcium ion diffusion in the tissue. In addition, NCPs may regulate collagen fibrillogenesis.

1-Carboxyglutamic Acid↗

PubMed1998

Two new in vitro calcification systems showing the higher calcifiability of enamel proteins than dentin and bone matrices.

One of the difficulties in simulating in vivo calcification by in vitro experiments is how to prepare and apply a suitable calcifying solution. We have previously developed an entirely new model system consisting of 40% acrylamide gel blocks that contains matrix proteins and is immersed in fetal calf serum at 37 degrees C. (40% gel system) for 18 hr (Connect. Tissue Res., 33, 185, 1995). The gels were analyzed for immobilized calcium. In this system bovine enamel proteins (0.1% in the gel) showed the highest calcifiability among the tested matrices, followed by insoluble bovine dentin, bone and skin collagens. The 40% gel system provides a barrier for high molecular weight inhibitor molecules in the body fluid. The new calcifying system developed in this study consists of the matrix protein sealed in dialysis tubing within a glass chromatography column that was eluted with a calcifying solution. In this system (dialysis tubing system), again the enamel protein showed higher calcifiability than dentin, bone and skin collagens. It was also shown that enamel proteins became not only a reversible opaque gel, but also a relatively-irreversible coagulant, if the solution contained calcium and phosphate ions at concentration below saturation (1 mM calcium and 1 mM phosphate). With both systems combined, deposition and crystal growth of minerals in enamel proteins will be better understood than with previous methods.

Acrylic Resins↗

PubMed1996

Relative dental development of Upper Pleistocene hominids compared to human population variation.

The relative development of permanent teeth in samples of Neandertal/archaic Homo and Early Modern/Upper Paleolithic hominids is compared to the range of variability found in three recent human samples. Both fossil hominid samples are advanced in relative M2 and M3 development compared to white French-Canadians, but only the Neandertal/archaic Homo M3 sample is advanced when compared to black southern Africans. Both fossil hominid samples are delayed in relative I1 and P3 development compared to the recent human samples. Two hypotheses concerning the significance of the advanced M3 and M2 development found in both hominid groups and southern Africans compared to French-Canadians are discussed. The first postulates that the differences in relative molar development are due simply to variation in tooth/jaw size relationships. The second postulates that the relatively advanced M3 and M2 development found in the fossil hominids and southern Africans is a correlate of their potential for advanced skeletal maturation compared to French-Canadians and other European-derived populations. It appears that dental development patterns have continued to evolve from the Upper Pleistocene to present times, and that Neandertals and Early Moderns shared similar patterns of relative dental development.

Animals↗

PubMed1996

Dental indicators of stress and reduced age at death in prehistoric Native Americans.

Considerable evidence supports the hypothesis that developmental enamel defects represent stress-induced growth disruptions. In this investigation, the relationship between different kinds of enamel defects and age at death is examined in the prehistoric Libben population from Ottawa County, Ohio. The sample consisted of the permanent dentitions of 143 individuals. Defects were classified based on the criteria of the Developmental Defects of Enamel (DDE) Index. The multifactorial age at death determinations of Lovejoy and coworkers (1977) were used in this analysis. Results reveal a significantly lower mean age at death for individuals with enamel defects vs. individuals with normal teeth. This pattern was clearly present for all defect types examined. No significant differences by sex were detected. The age-at-death distribution for individuals with normal teeth approximated the normal curve. The modal value was reached in the 35-40 year age class. The age-at-death distribution for individuals with enamel defects showed two peaks. The mode occurred in the 15-20 year age class, and the second, lower peak occurred in the 30-35 year age class. The early mortality of individuals with enamel defects may be related to biological damage to the immune system during prenatal or postnatal development.

Aging↗

PubMed1996

In vitro modeling of the bone/implant interface.

BACKGROUND: The purpose of this review is to examine the usefulness of cell culture methods to model the mechanisms of bone formation on the surfaces of candidate implant materials. METHODS: The central objective is to show that in vitro methods are uniquely valuable in providing an understanding of how new bone is formed on solid surfaces. It should be emphasized, at the outset, that the use of cell culture studies as cytotoxicity assays will not be addressed, nor is it implied that cell cultures can model all the complexities of the in vivo environment. Nevertheless, by comparison with in vivo data, which are by nature retrospective, it is shown that primary differentiating osteogenic cell cultures, derived from bone marrow, illustrate a sequence of extracellular matrix elaboration events that characterize the establishment of the interface between newly formed bone and solid surfaces. These solid surfaces either may be implant materials, or indeed previously formed bone matrix, which has been resorbed during normal bone remodeling events. In each case the first biologically derived matrix at these sites is a morphologically distinct collagen fibre-free extracellular matrix, which, in bone histology has been referred to for > 100 years as a cement line. RESULTS: The sequence starts with secretion and adsorption to the substratum of organic components, of which the major proteins are osteopontin and bone sialoprotein. Mineralization of this matrix occurs by the seeding of nanocrystalline calcium phosphate, which precedes the appearance of morphologically identifiable collagen fibres. This is clearly contrary to the dogma that collagen is necessary for mineralization of bone, but is in agreement with specific cases of other, particularly dental, calcified connective tissues. Although collagen is synthesized by the differentiating osteogenic cells that elaborate the cement line interface, it is not adsorbed to the underlying solid surface. Following the elaboration of the cement line matrix, collagen fibre assembly occurs and is then mineralized to produce morphologically identifiable bone matrix. CONCLUSION: Key elements of this sequence of events can be seen at the interface of implants retrieved from in vivo experiments, which indicates that these in vitro methods not only mimic known in vivo phenomena, but also provide a mechanistic understanding of bone elaboration at implant surfaces. However, distinction is drawn between the events of new bone formation at implant surfaces and other bone/implant morphologies, which are unrelated to de novo bone formation at the implant surface. Finally, this new information emerging from bone marrow cell culture studies demands a re-examination of the concepts of bone-bonding and nonbonding implant materials.

Adult↗

PubMed1978

Remineralization of enamel by a saliva substitute designed for use by irradiated patients.

A saliva substitute, VA-OraLube, was evaluated for ability to reharden dental enamel and to relieve intraoral soft tissue symptoms in patients receiving radiotherapy for malignancies of the head and neck. Treatments of 15, 30 and 60 minutes rehardened enamel by 3.1%, 4.0%, and 5.5%, respectively. In the second experiment, treatment for 60 minutes with the complete solution rehardened enamel by 5.2%. Omitting calcium, phosphorus and/or fluoride from the formulation greatly decreased this rehardening potential. Treatment of enamel with fresh whole saliva induced rehardening at a 7.3% level in comparison to the 5.5% and 5.2% derived by using the saliva substitute. Since the xerostomic patient usually uses the product very frequently, there is a remineralization potential of significant consequence. A total of 125 xerostomic patients used the saliva substitute on an ad lib basis over a period of 4 months. Patient responses indicated a very high level of acceptance and the virtual elimination of troublesome problems previously associated with the dry mouth state.

Head and Neck Neoplasms↗

PubMed1997

Tuftelin: enamel mineralization and amelogenesis imperfecta.

Tuftelin is a novel acidic enamel protein thought to play a major role in enamel mineralization. Its identity and localization has been confirmed by amino acid composition, enzyme-linked immunosorbant assay, Western blots, indirect immunohistochemistry and high resolution protein-A gold immunocytochemistry. The deduced tuftelin protein (pI 5.2) contains 389 amino acids and has a calculated peptide molecular mass of 43,814 Da. Immunological studies suggest conservation of tuftelin structure between species throughout vertebrate evolution. The cDNA sequence encodes for several putative post-translation sites including one N-glycosylation consensus site, seven O-glycosylation sites and seven phosphorylation sites, as well as an EF-hand calcium-binding domain (with mismatch), localized towards the N-terminal region. At the C-terminal region (residues 252-345) tuftelin contains structurally relevant determinants for self assembly. We recently cloned and partially sequenced the human tuftelin gene (four exons have now been sequenced). These sequences include exon 1 and over 1000 bases of the putative promoter region. Employing fluorescent in situ hybridization, we mapped the human tuftelin gene to chromosome 1q 21-31. Localization of the human tuftelin gene to a well-defined cytogenetic region may be important in understanding the aetiology of autosomally inherited amelogenesis imperfecta, the most common enamel hereditary disease.

Amelogenesis↗

PubMed1997

Enamel maturation.

Enamel maturation is characterized by massive crystal growth in both width and thickness, resulting in the most highly mineralized of all mammalian skeletal tissues. The control of this process is mediated via a carefully orchestrated series of events that are temporally and spatially regulated, and it requires the co-ordinated degradation and removal of the endogenous enamel matrix. This is affected by both neutral metalloproteases and serine proteases, which are developmentally restricted and may be further modulated by changes in the chemistry of the enamel crystals themselves. Failure of these mechanisms, or the adventitious entry of mineral-binding proteins during the later stages of maturation, may result in the incomplete maturation of the enamel crystals and the eruption of dysplastic tissue.

Albumins↗

PubMed1997

Structure and function of secretory ameloblasts in enamel formation.

Secretory ameloblasts have multiple functions including the synthesis and resorption of enamel matrix proteins and calcium transport during enamel formation. We have examined these functions by means of cytochemistry and immunocytochemistry. Enamel proteins, amelogenins and enamelins are localized in the biosynthetic pathways of ameloblasts and in the forming enamel. Sulfated glycoconjugates are present in secretory ameloblasts. The distal junctional complex of ameloblasts may act as a permeability barrier to enamel proteins, thereby confining the secreted proteins to the growing enamel front. Secretory ameloblasts contain lysosomal enzymes in the Golgi lysosome endoplasmic reticulum system and also exhibit absorptive capacity, which might be associated with an early decrease in extracellularly degraded enamel proteins. Active calcium transport through the ameloblasts towards the growing enamel is indicated by the demonstration of Ca-ATPase activity along the plasma membranes. A calcium-dependent modulator protein, calmodulin, is localized in ameloblasts, suggesting that early enamel mineralization is dependent upon calmodulin-regulated Ca-ATPase in ameloblasts. These results suggest that the secretory ameloblast is a highly specialized multifunctional cell in the production, resorption and degradation of enamel matrix and in the active calcium transport essential for matrix mineralization during enamel formation.

Ameloblasts↗