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Effects of thyro-parathyroidectomy and parathyroidectomy upon dentinogenesis: Part II: Electron microscopy.

An ultrastructural study was carried out in order to better characterize the findings observed in the first part of our study. The materials and methods are the same as those used in the preceding paper. This study reveals the occurrence of structures which display a symmetrical cross-banded pattern within the predentin and dentin of thyro-parathyroidectomized (TPTX) and parathyroidectomized (PTX) rats. A difference in the distribution of the symmetrical banded structures as dentinogenesis advances, as well as differences in the amount of the symmetrical banded structures between TPTX and PTX rats were observed. The symmetrical banded structures correspond with the so-called symmetrical SLS previously described in the incisor of normal and pathologic rats. The occurrence of these structures at a given stage of the incisor development suggests that the odontoblast is sensitive to the parathyroid hormone deficiency and/or hypocalcemia in a precise stage of its maturation.

Animals↗

Effects of vincristine on the developing hamster tooth germ in vitro.

Vincristine is one of the cytostatic drugs present in cocktails commonly used for the treatment of cancer in children. The aim of this study was to evaluate biochemically and histologically the toxic effects of this drug on the developing tooth in vitro using the organ culture model in order to be able to predict what damage the drug can induce in the developing teeth from children undergoing anti-neoplastic chemotherapy. The most profound effect of the drug (10(-8)M-10(-4)M vincristine) on the developing tooth germ was the induction of mitotic arrests at the cervical loop and in the inter-cuspal regions. The 10(-4)M-10(-6)M vincristine doses were cytotoxic to most cells in the developing tooth germ. The 10(-7)M vincristine dose apart from induction of mitotic arrests, did not appear to be cytotoxic to the mature differentiated secretory cells. However, this dose induced incomplete nuclear polarization of the differentiating ameloblasts and odontoblasts. At 10(-8)M vincristine, the only effect observed were mitotic arrests; the secretory cells did not appear to have been affected at all. On the other hand, mineralization (TCA-soluble 45Ca and 32P uptake) was dose-dependently decreased from 10(-7)M vincristine upwards. 10(-9)M vincristine, the lowest dose tested, did not induce any changes in the developing tooth germ. The organ culture data indicate that 10(-9)M vincristine is the highest (safe) dose which does not induce any toxic effects in the developing hamster tooth germ.(ABSTRACT TRUNCATED AT 250 WORDS)

Ameloblasts↗

Influence of substrate nature and immobilization of implanted dentin matrix components during induction of reparative dentinogenesis.

The biological effects of isolated soluble dentin extracellular matrix components on the induction of reparative dentinogenesis in exposed cavities in ferret canine teeth have been shown to be blocked by immobilizing the extracellular matrix components on nitrocellulose or Millipore membranes during implantation. This contrasts with the picture of induction of odontoblast-like cell differentiation and reparative dentin deposition on existing insoluble dentin matrix of the exposure walls when the extracellular matrix components are implanted in lyophilized form. These data indicate the importance of an existing insoluble dentin matrix in providing a substrate to potentiate the growth factor-like activity of soluble isolated dentin extracellular matrix components in the induction of reparative dentinogenesis.

Animals↗

Induction of reparative dentinogenesis in vivo: a synthesis of experimental observations.

EDTA--and/or guanidine HCl--insoluble dentinal matrix, or demineralized dentin which had been treated with plasma fibronectin, or pieces of Millipore filters coated with a recombinant fibronectin-like engineered polymer, incorporating many RGD sequences, were implanted into central parenchymal sites of young dog molars, via mechanical pulp exposures. Furthermore demineralized dentin and Millipore filters coated with plasma fibronectin were placed into the central pulp of old animals. Histological analysis of buffered formalin-fixed tissues showed that: 1. The dentinogenic activity was retained in the EDTA--and/or guanidine-insoluble dentin matrix. 2. Implantation of Millípore filters supplemented with the recombinant polymer did not induce any odontoblast-like cell differentiation, indicating that the interactions of pulp cells with the exogenous fibronectin are not RGD-dependent. 3. Acid-insoluble dentin matrix or plasma fibronectin (both separately inducing dentinogenesis in dental pulp of young animals) did not show any dentinogenic activity when exposed in pulp sites of old animals. Acid-insoluble dentin matrix and plasma fibronectin also failed to induce dentinogenic activity in the young pulpal tissues, when both factors were combined before to their implantation. Synthesizing the present data with previous relevant information it could be suggested that in the mechanism initiating reparative dentinogenesis, growth factors (endogenous or artificially implanted) and fibronectin are involved and this mechanism seems to be more complex than the simple immobilization of pulp cells onto an adhesion substratum.

Aging↗

Role of two mineral-associated adhesion molecules, osteopontin and bone sialoprotein, during cementogenesis.

Adhesion molecules and their cell membrane receptors are known to play important regulatory roles in cell differentiation. Consequently, the following experiments were conducted to determine the role of two adhesion molecules, bone sialoprotein (BSP) and osteopontin (OPN) in tooth root formation. Developing murine molar tooth germs at sequential stages of development (developmental days 21-42) were analyzed using immunohistochemical and in situ hybridization techniques. While BSP was localized to alveolar bone and odontoblasts early in development, BSP was distinctly localized to the cemental root surface at latter periods coincident with the initiation of root formation and cementogenesis. Conversely, OPN was distributed in a nonspecific fashion throughout the PDL and the eruption pathway of the forming tooth. In situ hybridization confirmed that cells lining the root surface express BSP. The fact that BSP is specifically localized to the cemental surface suggests that this protein is involved in cementoblast differentiation and/or early mineralization of the cementum matrix. Localization of OPN to non-mineralized tissues further suggests that OPN functions as an inhibitor of mineralization during periodontal ligament formation. These findings collectively suggest that BSP and OPN are intimately involved in the sequence of cellular and molecular events accompanying cementogenesis.

Alveolar Process↗

Models for the study of cementogenesis.

Cementum is a mineralized tissue that acts to connect the periodontal ligament to the tooth root surface. Its composition is very much like bone, being comprised mainly of type I collagen, inorganic mineral and noncollagenous proteins, however the origin of the cells and factors necessary for cementum formation have yet to be elucidated. Our laboratory has focused on the role that adhesion molecules, and their cell surface receptors, play in the formation of cementum and tooth root. In order to study this, we used a mouse molar as a model system. This system enabled us to study the formation of four distinct mineralized tissues; bone, cementum, dentin and enamel at various stages of their development. For these studies, we initiated experiments to examine potential cementoblast progenitor cells, in vitro. As a first step, we show that dental papilla and dental follicle cells, n vitro, obtained from molar tissues at day 21 of development, induce mineralized nodules, in vitro. In addition, we obtained tissues from mice where defects in root development may exist and determined bone sialoprotein (BSP) protein expression, a mineralized tissue specific adhesion molecule, in such tissues. As discussed here, we found that osteopetrotic (op/op) mice have delayed and/or defective root development and BSP does not localize in the dental tissues, at day 33 of development. In addition, dentin formation was defective and odontoblasts appeared immature, based on morphological examination. In contrast, the day 33 control molars demonstrated positive staining for BSP localized to root cementum, with normal formation of dentin.

Amelogenesis↗

Elemental distributions in predentine associated with dentine mineralization in rat incisor.

Electron probe microanalysis was applied to study quantitatively and semi quantitatively the elemental concentrations and distributions that occur in predentine during the dentine mineralization of rat incisor. Apex regions of the continuously growing incisors were rapidly dissected and cryofixed in liquid nitrogen-cooled propane. Ultrathin cryosections were prepared from the dentine tissue. On the average in the extracellular predentine element concentrations of calcium and phosphorus were about 0.5% (w/w) and 0.5-1% (w/w), respectively; so the calcium content in the extracellular predentine is higher while the phosphorus content is much lower than in the odontoblast area. Due to the high content of glycosaminoglycans in the extracellular matrix the concentration of sulfur in the predentine was more than 1% (w/w); the potassium content was found in the range of 0.6-0.8% (w/w) which is quite high for an extracellular area and the concentrations of sodium and chlorine were higher than 2% (w/w). Elemental mapping analysis was carried out to demonstrate the distribution of some important elements at the predentine/dentine border during mineralization.

Animals↗

Pulpal responses to various dentin bonding systems in dentin cavities.

The purpose of this study was to investigate pulpal response and bacterial invasion associated with five dentin bonding systems placed in cavities without an enamel margin. Two hundred and twenty-four dentin margin cavities were created by removing the surrounding marginal enamel of class V cavities in monkey teeth. These cavities were restored with a resin composite using various current dentin bonding systems. All of the dentin bonding systems showed odontoblastic layer changes to varying degrees, and slight to moderate inflammation at the initial stage. With the lapse of time, the degree of inflammation tended to decrease markedly, except for those cases in which bacteria were present. The result of SA/Photo Bond corresponded to that of the glass ionomer cement (negative control). Accordingly, this study suggests that a good bond and adaptation between resin and dentin should be obtained for eliminating pulpal irritation, even in the dentin cavity.

Animals↗

[Should the dentin smear layer be preserved or eliminated? (Review of the literature)].

The smear layer is a direct consequence of instrumentation of the root canal wall. Hand instrumentation as well as ultrasonic instrumentation produce a smear layer on the canal wall. This smear layer is composed of dentine, remnants of pulp tissue and odontoblastic processes and bacteria in an infected canal. Removal of the smear layer is accomplished by the irrigation of root canals with EDTA (17%) followed by NaOCl (5.25%). Permeability of the dentine is increased by the removal of smear layer. In this way the bacteria within the infected tubuli can be more easily destroyed by an intracanal dressing. Whether the removal of smear layer decreases the apical leakage is uncertain. To establish the clinical consequences from removal or preservation of the smear layer, further research is needed.

Bacteria↗

Histopathology of the pulp of primary molars with active and arrested dentinal caries.

The purpose of this study was to compare the histological appearance of the pulp of human primary molars with active and arrested lesions. The sample consisted of 36 primary molars (18 with active lesions and 18 with arrested lesions) extracted from 35 children between 5 to 9 years of age. The histological diagnosis was classified in normal pulp, transitional stage, partial pulpitis, total pulpitis and total necrosis, and then subdivided in three subgroups: treatable, untreatable and questionable. Results showed that normal pulp or transitional stage (treatable category) was diagnosed in 50% of teeth with arrested lesions, compared to 11.1% of teeth with active lesions. Partial pulpitis (questionable category) was present in 38.8% with arrested lesions compared to 22.2% with active lesions. Total pulpitis and total necrosis (untreatable category) was diagnosed in 11.2% with arrested lesions compared to 66.7% with active lesions. The observed frequencies of histological categories between both groups were statistically significant (P < 0.05). Histologically, pulp reaction under active and arrested lesions in primary molars revealed the formation of a basophilic calcio-traumatic line at the junction of the primary and reparative dentin, formation of reparative dentin and a regular odontoblastic layer in 60% of the cases. Results indicated that the type of lesion (active or arrested) is a good indicator of the histological status of the pulp.

Basophils↗

In situ hybridization analysis of transforming growth factor-beta 1 RNA expression during mouse tooth development.

OBJECTIVE: The aim of this study is to investigate the temporal and spatial expression and localization of transforming growth factor-beta 1 mRNA during mouse tooth development. METHODS: The distribution pattern of TGF-beta 1 mRNA during tooth development was analyzed on 5 microns serial sections of paraformaldehyde-fixed and paraffin-embedded embryonic mouse heads or mandibles. RESULTS: The results show that TGF-beta 1 mRNA is expressed during mouse tooth development in a temporally and spatially regulated fashion. Local expression of TGF-beta 1 mRNA in the dental epithelium at budstaged (E13) and capstaged (E15) teeth were observed. During bell stage (E16-18), TGF beta 1 mRNA was very abundant in the ameloblast layer and dental papilla cells. The expression of TGF-beta 1 increased in the layer of odontoblast and ameloblast with the differentiation of these cells. CONCLUSION: These results suggest that TGF-beta 1 may have an important role in odontogenesis and that it acts as a paracrine and autoinducing factor.

Ameloblasts↗

Immunohistochemistry of bone sialoprotein and osteopontin during reparative dentinogenesis in vivo.

OBJECTIVE: To investigate the stage-specific and tissue expression of bone sialoprotein (BSP) and osteopontin (OPN) during reparative dentinogenesis in vivo. METHODS: Direct pulp-capping with Ca(OH)2 developed a model used for the investigation of reparative dentinogenesis in the exposed dental pulp. In this model, standardized class V cavities were prepared close to the gingival margin on the buccal surface of each tooth. Animals were sacrificed 3, 7, and 10 days post-operation. Immunohistochemical staining determined the tissue-specific expression of BSP and OPN during the process. RESULTS: Odontoblast-like cells and reparative dentin reacted positively with BSP and OPN. The expression of BSP reached a peak at day 7, then gradually decreased, while the expression of OPN showed no significant difference between day 7 and day 10. CONCLUSION: BSP and OPN may play different roles in reparative dentinogenesis. BSP may serve as a nucleator of hydroxyapatite crystal formation.

Animals↗

[Electroanalgesia: operating principles].

We are faced every day with the problem of dental pain arising from the concomitant action of the nerve fibres, the odontoblasts and the dentinal fluid. Its propagation is due to the triggering of the action potential from the resting potential. It can be reduced thanks to a reversible process of anelectrotonus in electro-analgesia.

Action Potentials↗

Self-contamination of deep dentin by dentin fluid.

PURPOSE: To compare, with the use of a resin replica technique, surface features of deep, acid-conditioned dentin from vital human molars that were anesthetized with an anesthetic: Group 1: without a vasoconstrictor (Mepivacaine 3%), and Group 2: containing a vasoconstrictor (Lidocaine 2% with 1:80,000 epinephrine). MATERIALS AND METHODS: 10 Class I cavities with dentin caries were included in each group. Following complete caries removal, a total-etch technique was performed with 32% phosphoric acid (Uni-Etch) for 15 s. After rinsing, each cavity was air-dried for 1 s, then a slow setting vinyl polysiloxane (President) impression was taken. As a control, impressions were taken from three additional cavities in teeth that were anesthetized with Mepivacaine 3% but not acid etched. A TEM-grade epoxy resin was used to prepare replicas from the impressions. Polymerized replicas were coated with gold and examined with a scanning electron microscope. RESULTS: In Group 1, fluid was visible leaving tubular orifices in all replicas. In addition, three were covered with a smooth, amorphous film that was different from the granular appearance of the smear layer in the control cavities. In Group 2, patent tubular orifices without fluid were observed in all specimens. Odontoblast process-like structures were seen from some dentin tubules.

Acid Etching, Dental↗

Pulp response to direct capping with an adhesive system.

PURPOSE: To evaluate the pulp response following direct pulp capping with an adhesive system (Prime & Bond 2.0 - PB 2.0) and a zinc-oxide eugenol cement (ZOE) on pulp exposures in rat molar teeth. MATERIALS AND METHODS: Forty-eight Class I cavities were prepared on the occlusal surface of molar teeth of rats (Rattus Norvegicus, Holtzman). Pulp exposures performed on the cavity floor were capped either with the adhesive system P&B 2.0 or ZOE. After 7, 15, 30, and 60 days, the specimens were processed through H & E and Brown & Brenn staining techniques. RESULTS: Both pulp capping materials allowed pulp repair, characterized by reorganization of a new odontoblast cell layer underlying the dentin bridge formation. However, P&B 2.0 promoted a large zone of cell-rich fibrodentin matrix deposition between the pulp capping material and the dentin bridge, which was deposited far from the pulp exposure site. On the other hand, pulps capped with ZOE showed dentin bridging immediately subjacent to the pulp capping material. In those samples in which microleakage occurred between dental material and cavity walls there was a persistent inflammatory reaction and lack of complete pulp repair.

Animals↗

Cytotoxic effects of cleansing solutions recommended for chemical lavage of pulp exposures.

PURPOSE: To evaluate the in vitro cytotoxic effects of three cleansing solutions used for chemical lavage of pulp exposures. MATERIALS AND METHODS: The immortalized odontoblast cell line (MDPC-23) was plated (30,000 cells/cm2) and incubated for 72 hrs in 24-well dishes. After counting the cell number under inverted light microscopy, 20 microl of the experimental and control solutions were added to 980 microl of fresh culture medium. Then, hydrogen peroxide (3%, H2O2), sodium hypochlorite (6%, NaOCl) or calcium hydroxide-saline solution (5g of Ca(OH)2 in 10 ml of sterile distilled water) were added to wells for experimental Groups 1, 2 and 3, respectively. The positive and negative control groups received Syntac Sprint bonding agent (SS) and phosphate buffered saline (PBS), respectively. Following incubation for 120 min the cell number was counted again, the cell morphology was evaluated by scanning electron microscopy (SEM) and the cell metabolism was determined by the methyltetrazolium (MTT) assay. The scores obtained from cell counting and MTT assay were analyzed with an ANOVA followed by Fisher's PLSD tests. RESULTS: H2O2, NaOCl solutions, and SS bonding agent were more cytotoxic than Ca(OH)2 or PBS. In the groups with H2O2 or SS, only a few cells remained attached to the bottom of wells. The difference between these two groups was not statistically significant. H2O2, NaOCl and SS depressed the mitochondrial enzyme response by 97.7%, 97.3%, and 95.0%, respectively. On the other hand, Ca(OH)2 depressed the metabolic activity of cells by only 5%. While H2O2, NaOCl and SS caused extreme changes on the cell morphology, neither Ca(OH)2 nor PBS promoted dramatic changes in the cell morphology.

Analysis of Variance↗

Impairment of molar tooth eruption caused by x-radiation.

The effect of x-radiation on erupting molars is presented. New born, 5 day old Wistar rats were locally irradiated in the molar area with doses of 20 Gy. They were killed in two groups, 30 and 60 days postirradiation respectively. Two other groups of non irradiated, age matched rats were killed at the given times. In addition a control group of 5 day old animals was also studied. Radiographic and histologic studies were performed. Odontoblastic atrophy, odontodysplasia, rootless formation, and ankylosis of tooth to bone by osteodentin formation with the resulting lack of tooth eruption were observed. The relation between the histologic alterations and tooth eruption is discussed.

Animals↗

Pulpal response to a fluoride-releasing all-in-one resin bonding system.

Pulp tissue reactions to a fluoride-releasing all-in-one resin bonding system (Reactmer Bond and Reactmer Paste) in non-exposed monkey teeth were histopathologically evaluated at three, 30, and 90 days after restoration. No serious inflammatory reactions of the pulp, such as necrosis or abscess formation, were observed. At 90 days in the Reactmer group, odontoblastic change and inflammatory cell infiltration were not observed, and slight irritation dentin formation was formed. The pulpal response of the Reactmer group was minimally different from that of the control group. Consequently, the Reactmer system was determined as being biologically compatible with vital pulps.

Abscess↗