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1.
J Dent Res ; 86(5): 463-8, 2007 May.
Article in English | MEDLINE | ID: mdl-17452569

ABSTRACT

Growth hormone (GH) status affects dental development, but how GH influences tooth size/shape is unclear. Since GH affects dental epithelial proliferation, we hypothesized that GH influences the tooth crown and root dimensions. Dentin matrix dimensions were measured in longitudinal sections of decalcified first mandibular molars from 3 genetically modified mice: giant (GH-Excess) mice and dwarf (GH-Antagonist and GH-Receptor-Knockout) mice. GH status was found to influence crown width, root length, and dentin thickness. Analysis of these data suggests that GH influences both tooth crown and root development prior to dentinogenesis as well as during appositional growth of dentin. This is concordant with the expression of paracrine GH and GH receptors during tooth bud morphogenesis, and of GH receptors in the enamel organ, dental papilla, and Hertwig's epithelial root sheath during dentinogenesis. Based on prior studies, these GH morphogenetic actions may be mediated by the induction of both bone morphogenetic protein and insulin-like growth factor-1 expression.


Subject(s)
Dentin/anatomy & histology , Growth Hormone/physiology , Odontogenesis/physiology , Animals , Dentinogenesis/physiology , Female , Growth Hormone/pharmacology , Male , Mice , Mice, Knockout , Mice, Transgenic , Molar/anatomy & histology , Recombinant Proteins/pharmacology , Tooth Crown/anatomy & histology , Tooth Root/anatomy & histology
2.
J Dent Res ; 83(1): 35-9, 2004 Jan.
Article in English | MEDLINE | ID: mdl-14691110

ABSTRACT

Cementum is known to be growth-hormone (GH)-responsive, but to what extent is unclear. This study examines the effects of extremes of GH status on cementogenesis in three lines of genetically modified mice; GH excess (giant), GH antagonist excess (dwarf), and GH receptor-deleted (GHR-KO) (dwarf). Age-matched mandibular molar tissues were processed for light microscope histology. Digital images of sections of first molar teeth were captured for morphometric analysis of lingual root cementum. Cross-sectional area of the cellular cementum was a sensitive guide to GH status, being reduced nearly 10-fold in GHR-KO mice, three-fold in GH antagonist mice, and increased almost two-fold in giant mice (p < 0.001). Cellular cementum length was similarly influenced by GH status, but to a lesser extent. Acellular cementum was generally unaffected. This study reveals cellular cementum to be a highly responsive GH target tissue, which may have therapeutic applications in assisting regeneration of the periodontium.


Subject(s)
Dental Cementum/pathology , Growth Hormone/physiology , Animals , Cementogenesis/physiology , Dwarfism/genetics , Dwarfism/physiopathology , Female , Gigantism/genetics , Gigantism/physiopathology , Image Processing, Computer-Assisted , Male , Mice , Mice, Inbred BALB C , Mice, Inbred C57BL , Mice, Inbred Strains , Mice, Knockout , Mice, Transgenic , Molar/pathology , Odontometry , Phenotype , Receptors, Somatotropin/genetics , Receptors, Somatotropin/physiology , Tooth Root/pathology
3.
Eur J Oral Sci ; 109(4): 260-6, 2001 Aug.
Article in English | MEDLINE | ID: mdl-11531072

ABSTRACT

A body of published evidence suggests that a significant portion of enamel matrix protein synthesized by ameloblasts localises in the lysosomal-endosomal organelles of these enamel organ cells. Little is known regarding the lysosomal proteolytic activities during amelogenesis. The aims of this study were to detect and measure the activities of lysosomal peptidases cathepsin B (E.C. 3.4.22.1) and dipeptidyl-peptidase II (E.C. 3.4.14.2) in the enamel organ of the rat incisor and to ascertain whether rat enamel matrix proteins are degraded by these peptidases in vitro. Whole enamel organs were dissected from rat mandibular incisors. Enamel protein was also collected from the rat teeth. Analysis indicated that the rat incisor enamel organs contained specific activities of both dipeptidyl-peptidase II and cathepsin B at levels comparable with those of kidney which is rich in both these lysosomal peptidases. Gel electrophoresis and immunoblotting demonstrated that both cathepsin B and dipeptidyl-peptidase II were able to substantially degrade the rat enamel proteins in vitro. Based on these observations, we propose that lysosomal proteases have roles in amelogenesis in the intracellular degradation of amelogenins.


Subject(s)
Amelogenesis/physiology , Cathepsin B/metabolism , Dipeptidyl-Peptidases and Tripeptidyl-Peptidases/metabolism , Enamel Organ/enzymology , Amelogenin , Animals , Blotting, Western , Cathepsin B/analysis , Chromogenic Compounds , Coumarins , Dental Enamel Proteins/analysis , Dental Enamel Proteins/metabolism , Dipeptides , Dipeptidyl-Peptidases and Tripeptidyl-Peptidases/analysis , Electrophoresis, Polyacrylamide Gel , Fluorescent Dyes , Hydrogen-Ion Concentration , Immunoblotting , Incisor , Kidney/enzymology , Lysosomes/enzymology , Rats , Rats, Wistar , Spectrometry, Fluorescence , Statistics as Topic
4.
Apoptosis ; 4(6): 441-7, 1999 Dec.
Article in English | MEDLINE | ID: mdl-14634328

ABSTRACT

Enamel-producing cells (ameloblasts) pass through several phenotypic and functional stages during enamel formation. In the transition between secretory and maturation stages, about one quarter of the ameloblasts suddenly undergo apoptosis. We have studied this phenomenon using the continuously erupting rat incisor model. A special feature of this model is that all stages of ameloblast differentiation are presented within a single longitudinal section of the developing tooth. This permits investigation of the temporal sequence of gene and growth factor receptor expression during ameloblast differentiation and apoptosis. We describe the light and electron microscopic morphology of ameloblast apoptosis and the pattern of insulin-like growth factor-1 receptor expression by ameloblasts in the continuously erupting rat incisor model. In the developing rat incisor, ameloblast apoptosis is associated with downregulated expression of the insulin-like growth factor-1 receptor. These data are consistent with the hypothesis that ameloblasts are "hard wired" for apoptosis and that insulin-like growth factor-1 receptor expression is required to block the default apoptotic pathway. Possible mechanisms of insulin-like growth factor-1 inhibition of ameloblast apoptosis are presented. The rat incisor model may be useful in studies of physiological apoptosis as it presents apoptosis in a predictable pattern in adult tissues.

5.
Anat Rec ; 233(4): 493-503, 1992 Aug.
Article in English | MEDLINE | ID: mdl-1626709

ABSTRACT

Dipeptidyl peptidase II (DPP II), E.C. 3.4.14.2, a serine class endopeptidase, is widely used as a lysosomal marker in cytochemical studies. To date most ultrastructural studies of ameloblasts use the presence of acid phosphatase activity to identify cellular organelles to be lysosomal. Using decalcified rat mandibles, with kidney tissue as a positive control, DPP II activity, was assessed with specific substrate Lysyl-alanine-4-methoxy-2-naphthylamide in ameloblasts at an ultrastructural level. Reaction product (RP) indicative of DPP II activity was observed only within lysosome-like organelles. These RP-labelled organelles were only localized in the supra- or para-nuclear regions of the ameloblasts, which corresponds with previous studies using acid phosphatase cytochemistry. However, in contrast with these studies, RP was not detected in the distal region of the ameloblasts, viz., in the Tomes' processes of the secretory ameloblasts or near the ruffled border in the maturation ameloblasts. The transitional ameloblasts were notable for the intensity of staining of their RP-labelled organelles. We propose that DPP II may have a role in programmed cell death which is thought to occur in this transition zone. Biochemical analysis of rat incisor enamel organ homogenates, indicated tissue fixation resulted in an 82% reduction in DPP II activity, although the specific activity of DPP II was not affected.


Subject(s)
Ameloblasts/enzymology , Dental Enamel/enzymology , Dipeptidyl-Peptidases and Tripeptidyl-Peptidases/metabolism , Lysosomes/enzymology , Ameloblasts/ultrastructure , Amelogenesis , Animals , Biomarkers , Dental Enamel/cytology , Dental Enamel/ultrastructure , Histocytochemistry , Incisor , Male , Microscopy, Electron , Rats , Rats, Inbred Strains
6.
Arch Oral Biol ; 35(8): 671-5, 1990.
Article in English | MEDLINE | ID: mdl-2256823

ABSTRACT

Female Wistar rats, 3 weeks old, were given sodium fluoride in saline solution (isotonic) by intraperitoneal injection at a dose of either 0, 10 or 20 mg per kg body weight. This treatment was given 9 times over 4.5 days. After fixation by perfusion and demineralization in neutral EDTA, hemi-mandibles were sectioned in a cryostat. Sections were stained for dipeptidyl peptidase II activity, using the specific substrate Lys-Ala-MNA and the coupler Fast Blue B for histochemical localization. Staining indicative of dipeptidyl peptidase II was found in the enamel organ of the incisor, particularly in cells of the stratum intermedium and in both secretory and maturation ameloblasts. This staining was markedly reduced in ameloblasts of rats given either 10 or 20 mg sodium fluoride per kg body weight.


Subject(s)
Ameloblasts/enzymology , Dipeptidyl-Peptidases and Tripeptidyl-Peptidases/metabolism , Enamel Organ/cytology , Sodium Fluoride/pharmacology , Acid Phosphatase/metabolism , Ameloblasts/drug effects , Amelogenesis , Animals , Enamel Organ/enzymology , Female , Incisor , Rats , Rats, Inbred Strains , Sodium Fluoride/administration & dosage
7.
Calcif Tissue Int ; 41(2): 105-11, 1987 Aug.
Article in English | MEDLINE | ID: mdl-3115544

ABSTRACT

Various doses of sodium fluoride (NaF), salmon calcitonin (CT) and NaF combined with CT were given intravenously to rats (236 +/- 2 g). Blood plasma samples were collected at various times up to 24 hours and measured for total calcium (adjusted for variation in plasma albumin), inorganic phosphate, ionic fluoride, urea, and creatinine. Following injection of NaF alone, significant hypocalcemia and hyperphosphatemia was observed. In contrast, CT injections resulted in hypocalcemia and hypophosphatemia. NaF and CT given in combination generally resulted in the hypocalcemia being equal to or greater than that calculated from the simple addition of their individual effects. Significant increases in both plasma creatinine and plasma urea were observed following treatment with NaF alone or with CT and NaF together, whereas CT alone had negligible effect. These results suggest that NaF does not mimic the effects of CT; rather that NaF and CT interact to modify their individual effects. The influence of NaF probably occurs via an effect on kidney function.


Subject(s)
Calcitonin/pharmacology , Calcium/blood , Fluorides/blood , Phosphates/blood , Sodium Fluoride/pharmacology , Analysis of Variance , Animals , Creatinine/blood , Male , Rats , Rats, Inbred Strains , Urea/blood
8.
J Dent Res ; 64(11): 1281-5, 1985 Nov.
Article in English | MEDLINE | ID: mdl-3867685

ABSTRACT

This study assessed the effect of a single intravenous dose of sodium fluoride (20 mg/kg body wt.) on plasma ionic fluoride and on some other plasma electrolytes and metabolites in rats, rabbits, and cockerels. At any given time following sodium fluoride administration, the plasma ionic fluoride was highest in rabbits and lowest in cockerels. The rate of removal of fluoride from plasma was slower in rabbits as compared with that in the other two species. Plasma sodium, chloride, total protein, albumin, total globulins, and osmolality were not significantly altered by sodium fluoride in any of these three species. However, plasma phosphate (inorganic), urea, creatinine, and glucose were elevated, and plasma calcium was reduced in the rats and the rabbits, but none was significantly altered in the cockerels. The analyses indicated that species variability does exist in fluoride toxicity.


Subject(s)
Electrolytes/blood , Sodium Fluoride/administration & dosage , Animals , Blood Glucose/analysis , Calcium/blood , Chickens , Creatinine/blood , Male , Potassium/blood , Rabbits , Rats , Rats, Inbred Strains , Sodium Chloride , Sodium Fluoride/blood , Sodium Fluoride/pharmacology , Urea/blood
10.
J Dent Res ; 63(8): 1061-3, 1984 Aug.
Article in English | MEDLINE | ID: mdl-6430985

ABSTRACT

EDTA-soluble material, extracted from incisor teeth of rats given 50 ppm fluoride (NaF) in drinking water ad libitum from conception to age nine wk, contained significantly higher levels of protein (P less than 0.01) and gamma-carboxyglutamic acid (P less than 0.01) than did similar material from control rats. Tooth fluoride levels indicated that the incisors were mildly fluorosed. Analysis of these data indicates that mild fluorosis of rat incisor teeth involves not only the characteristic disturbances of pigmentation and mineral structure but also some alteration of the matrix protein.


Subject(s)
1-Carboxyglutamic Acid/analysis , Fluorides/pharmacology , Glutamates/analysis , Incisor/analysis , Proteins/analysis , Amino Acids/analysis , Animals , Edetic Acid , Female , Fluorides/administration & dosage , Fluorides/analysis , Fluorosis, Dental/metabolism , Maternal-Fetal Exchange , Pregnancy , Rats , Rats, Inbred Strains , Solubility
11.
Calcif Tissue Int ; 30(1): 57-66, 1980.
Article in English | MEDLINE | ID: mdl-6153923

ABSTRACT

Over a 24-h period, Wistar rats from 4 litters, 6 to 9 days old, were given five intraperitoneal injections of a solution of 0.9% sodium chloride containing sodium fluoride (3 mg F/kg body weight). Within-litter controls were used. All rats were killed by decapitation 2 h after the final injection and the rat heads, cut sagittally, were processed for protein histochemistry. The intensity of staining of the protein in the enamel matrix of the upper jaw molar tooth buds was quantified using the two-wavelength method of microphotometry. A significant increase in the intensity of staining of fluoride-treated tissues over controls was observed with the histochemical methods specific for arginine (P less than 0.01), tyrosine (P less than 0.05), and cysteine (P less than 0.05). Other histochemical methods specific for amino acid groups failed to show any significant difference between fluoride and non-fluoride-treated enamel matrix.


Subject(s)
Dental Enamel Proteins/metabolism , Dental Enamel/drug effects , Fluorides/pharmacology , Tooth Germ/drug effects , Animals , Arginine/metabolism , Dental Enamel/anatomy & histology , Dental Enamel Proteins/analysis , Histocytochemistry , Maxilla , Molar , Rats , Spectrophotometry/methods , Staining and Labeling , Sulfhydryl Compounds/metabolism , Tyrosine/metabolism
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