The cervical wedge-shaped lesion in teeth: a light and electron microscopic study.

BACKGROUND: The cervical non-carious wedged-shaped lesion is controversial in that its aetiology may involve attrition, erosion, abrasion and stress-corrosion (abfraction). This study examined the histopathology of anterior teeth with cervical wedge-shaped lesions by light and electron microscopy to elucidate their pathogenesis. METHODS: Ten undecalcified human teeth with cervical lesions were available for investigation. Patency of the dentine tubules was tested using red dye penetration from the pulp chamber. The morphology of normal and sclerotic dentine adjacent to the cervical wedge-shaped lesions was investigated by scanning electron microscopy. The numbers and diameters of dentinal tubules were measured at different levels beneath the surfaces of the lesions. RESULTS: The gross and microscopic features of the worn teeth were described. Red dye penetration tests showed white tracts of sclerotic tubules contrasted with red tracts of patent tubules. Numbers of tubules per square area and diameters of patent and sclerotic tubules varied at different levels within the dentine due to deposits of intratubular dentine. CONCLUSIONS: The cervical wedge is shaped by interactions between acid wear, abrasion and dentinal sclerosis. No histopathological evidence of abfraction was found. Clinical diagnosis, conservation and restoration of non-carious cervical lesions need to take into account the extent of sclerotic dentine beneath wedge-shaped lesions.

Endodontic sequelae of dental erosion.

BACKGROUND: The incidence of pulp involvement in patients with excessive wear has not been extensively documented. METHODS: Clinical records of 448 patients with excessive tooth wear were reviewed and 52 cases (11.6 per cent) with near or frank pulp exposures or root canal treatments were found and their numbers and sites were tabulated. Light microscopy of study models was used to determine aetiology at each site of exposure as attrition, erosion or abrasion, scanning electron microscopy (SEM) was performed on some individual teeth. RESULTS: Forty sites of near exposure and 57 sites of frank exposures or root canal treatments were found, some cases had both types of exposure. The commonest sites exposed by erosion were the palatal surfaces of maxillary, and the incisal surfaces of mandibular anterior teeth. Posterior teeth were not commonly affected. Toothbrush abrasion had exacerbated some lesions as shown by SEM. CONCLUSIONS: Endodontic sequelae were found in 11 per cent of tooth wear patients as late stages of dental erosion. Near and frank exposures of the pulp thus constitute a small but significant, problem for the Australian dental profession's concern in the management of the tooth wear cases.

Effect of ethylene vinyl acetate (EVA) closed cell foam on transmitted forces in mouthguard material.

OBJECTIVES: To compare transmitted forces through ethylene vinyl acetate (EVA) mouthguard material and the same EVA material with gas inclusions in the form of a closed cell foam. METHOD: EVA mouthguard materials with and without foam gas inclusions and 4 mm thick were impacted with a constant force from an impact pendulum. Various porosity levels in the foam materials were produced by 1%, 5%, and 10% by weight foaming agent. The forces transmitted through the EVA after energy absorption by the test materials were measured with a force sensor and compared. RESULTS: Only minor non-significant differences in transmitted forces through the EVA with and without foam were shown. CONCLUSIONS: The inclusion of gas in the form of a closed cell foam in 4 mm thick EVA mouthguard materials did not improve the impact performance of the EVA mouthguard material.

Dental erosion in asthma: a case-control study from south east Queensland.

BACKGROUND: Asthma medication places patients at risk of dental erosion by reducing salivary protection against extrinsic or intrinsic acids. But patterns of lesions in asthmatics may differ from patterns in non-asthmatics, because gastro-oesophageal reflux (GOR) is found in 60 per cent of asthmatics. METHODS: The lesions in 44 asthma cases were compared to those of age and sex match controls with no history of asthma or medications drawn from the dental records of 423 patients referred concerning excessive tooth wear. The subjects were 70 males age range 15 to 55 years and 18 females age range 18 to 45. Anamnestic clinical data were compared between the two groups. Models of all 88 subjects were examined by light microscopy, and wear patterns were recorded on permanent central incisor, canine, premolar and first molar teeth. RESULTS: Clinical differences were a higher incidence of tooth hypersensitivity, xerostomia, salivary gland abnormalities, gastric complaints, and self induced vomiting in the cases. No differences were found between the cases and controls on citrus fruit and acid soft drink consumption. More occlusal erosion sites were found in cases, whereas more attrition sites were found in the controls. There were no significant differences in palatal erosion on maxillary anterior teeth found between cases and controls. Lingual erosion of the mandibular incisors, found only in GOR patients, was not observed. CONCLUSIONS: A higher incidence of erosion was found in asthmatics. Gastro-oesophageal reflux symptoms were not associated with the sign of lingual mandibular incisor erosion. The clinical significance is that asthmatics are at risk of dental erosion from extrinsic acid, but GOR does not appear to contribute in a site-specific manner.

Effect of a static magnetic field on orthodontic tooth movement in the rat.

Orthodontic tooth movement may be enhanced by the application of a magnetic field. Bone remodelling necessary for orthodontic tooth movement involves clastic cells, which are tartrate-resistant acid phosphatase (TRAP) positive and which may also be regulated by growth hormone (GH) via its receptor (GHR). The aim of this study was to determine the effect of a static magnetic field (SMF) on orthodontic tooth movement in the rat. Thirty-two male Wistar rats, 9 weeks old, were fitted with an orthodontic appliance directing a mesial force of 30 g on the left maxillary first molar. The appliance incorporated a weight (NM) or a magnet (M). The animals were killed at 1, 3, 7, or 14 days post-appliance insertion, and the maxillae processed to paraffin. Sagittal sections of the first molar were stained with haematoxylin and eosin (H&E), for TRAP activity or immunohistochemically for GHR. The percentage body weight loss/gain, magnetic flux density, tooth movement, width of the periodontal ligament (PDL), length of root resorption lacunae, and hyalinized zone were measured. TRAP and GHR-positive cells along the alveolar bone, root surface, and in the PDL space were counted. The incorporation of a SMF (100-170 Gauss) into an orthodontic appliance did not enhance tooth movement, nor greatly alter the histological appearance of the PDL during tooth movement. However significantly greater root resorption (P = 0.016), increased width of the PDL (P = 0.017) and greater TRAP activity (P = 0.001) were observed for group M at day 7 on the compression side. At day 14 no differences were observed between the appliance groups.

The surface effect of dentifrices.

The aim of this study was to evaluate clinically three commercially available dentifrices and to determine any surface effects on tooth or gingival surfaces. Sixty-four participants were included in this study and were allocated randomly to one of four treatment groups by an independent person to ensure the investigators were unaware of the brushing material used. All toothbrushes and dentifrices were distributed by this independent person. The treatment groups were: Group 1--brush with water; Group 2--brush with Colgate (Baking Soda and Peroxide); Group 3--brush with Macleans (Whitening); Group 4--brush with Colgate (Sensation Whitening). All participants were requested to brush both morning and evening in their customary fashion using only the designated toothpaste, or water, for four weeks. All participants were required to use the same toothbrush type. No other oral hygiene products such as mouth rinses or dental floss were used during the trial period. Prior to commencement of the brushing period, all participants received a full clinical examination recording the status of the soft and hard tissues including a gingival index (Löe and Silness) to record gingival condition. A polyvinyl siloxane impression was taken of the six anterior teeth and gingival tissues at the commencement of the trial. After four weeks, a second full clinical examination was made and further silicone impressions were taken of the anterior teeth. All impressions were cast in epoxy resin for investigation with light and electron microscopy. Participants were also asked to answer a questionnaire relating to the toothpaste used. The results of this study indicated that no significant clinical differences were recorded for any dentifrice or water and there was no significant difference in gingival index scores over the four week period. Patient responses to each dentifrice varied according to individual patient preferences and expectations and no consistent findings could be determined. Light and electron microscopy indicated that tooth and gingival surface changes that occurred over the four week period with any of the dentifrices were similar to, and not significantly different from, changes seen with the use of water alone. These results indicate that none of the dentifrices tested was harmful to teeth or soft tissues.

Orthodontic tooth movement in the prednisolone-treated rat.

Adverse effects of corticosteroids on bone metabolism raise concerns as to whether steroid treatment may influence orthodontic movement. This study examined the effect of prednisolone on orthodontic movement using an established rat model. The corticosteroid treated group (N = 6) was administered prednisolone (1 mg/kg) daily, for a 12-day induction period; the control group (N = 6) received equivalent volumes of saline. On day 12, an orthodontic appliance was placed which exerted 30 g of mesial force to the maxillary first molar. Animals were sacrificed on day 24 and tooth movement was measured. Sagittal sections of the molars were stained with haematoxylin and eosin, and for tartrate-resistant acid phosphatase (TRAP) activity. While there were no significant differences in the magnitude of tooth movement between the 2 groups, steroid-treated rats displayed significantly less root resorption on the compression side and fewer TRAP-positive cells within the PDL space on the same side. This suggests steroid treatment suppressed clastic activity.

The effect of glucocorticosteroid treatment on dentine formation in the Lewis rat, a histological study.

Glucocorticosteroids are widely used in the treatment of chronic illnesses and have been reported to cause premature obliteration of the pulp space. During the active stages of dentinogenesis, odontoblasts are growth hormone receptor (GHr) positive. The aims of this study were to determine if the glucocorticosteroid, prednisone, affected the rate of dentine deposition and odontoblast expression of GHr in the rat molar. Following subcutaneous injection of 0.05 mg/kg, 1.0 mg/kg or 5.0 mg/kg prednisone for 20 days, immature and mature molars from rats aged 3 and 6 weeks respectively, were examined histologically. Distribution of GHr expression was determined immunohistochemically. No morphological differences were observed in molars from prednisone treated animals. Prednisone did not appear to enhance dentine deposition in immature molars but in mature molars significantly increased dentine deposition on the roof of the pulp chamber at a dosage of 5.0 mg/kg (p < 0.001). In all immature molars, odontoblasts and pulp cells expressed GHr immunoreactivity. In mature molars, odontoblasts and pulpal cells from controls did not show GHr immunoreactivity. However, odontoblasts and pulp cells were GHr immunoreactive in mature molars from animals treated with prednisone.

Ameloblast apoptosis and IGF-1 receptor expression in the continuously erupting rat incisor model.

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.

An assessment of the osteoinductive potential of commercial demineralized freeze-dried bone in the murine thigh muscle implantation model.

Early studies have demonstrated that implantation of laboratory preparations of demineralized freeze dried bone (DFDB) into the thigh muscle of mice induces ectopic osteoinduction. However, with the development of commercial preparations of DFDB for clinical use, concerns have been raised as to the osteoinductive properties of such preparations. The aim of this study was to investigate the osteoinductive potential of some commercial preparations of DFDB compared to a newly developed product which incorporates DFDB into a collagen sponge. Commercial preparations of DFDB or the DFDB/collagen sponge were inserted into the thigh muscles of 60 adult Swiss CD-1 mice. At the completion of each experimental period (7, 14, 30, 90 and 180 days), the animals were sacrificed, and the hindquarters of the mice were radiographed. The area where each graft had been placed was then excised, processed for light microscopy, and stained with hematoxylin and eosin or von Kossa's stain. Histological analysis of the DFDB/collagen sponges demonstrated significant remineralization which increased with time. Remineralization of the DFDB/collagen sponges was verified by radiographs which showed a significant increase in radiopacity over time. There was no radiographic evidence of mineralized tissue formation or remineralization in any of the commercial DFDB samples studied. At all time points studied, histological analyses failed to show evidence of bone formation for any of the preparations. The results suggest that commercially available DFDB is not osteoinductive in the murine model and question the use of such materials in clinical periodontics. The results found for the DFDB/collagen sponge indicate a different mechanism of activity from DFDB as evidenced by its rapid remineralization. The role this remineralization process has in osteoinduction is unknown and requires further study.

Non-expression of insulin-like growth factor-I receptor is associated with apoptosis: an ultrastructural study on rat ameloblasts.

Insulin-like growth factor-I (IGF-I) is a pleiotrophic polypeptide which appears to have roles both as a circulating endocrine hormone and as a locally synthesized paracrine or autocrine tissue factor. IGF-I plays a major role in regulating the growth of cells in vivo and in vitro and initiates metabolic and mitogenic processes in a wide variety of cell types by binding to specific type I receptors in the plasma membrane. In this study, we report the distribution of IGF-I receptors in odontogenic cells at the ultrastructural level using the high resolution protein A-gold technique. In the pre-secretory stage, very little gold label was visible over the ameloblasts and odontoblasts. During the secretory stage the label was mostly seen in association with the cell membranes and endoplasmic reticulum of the ameloblasts. Lysosome-like elements in the post-secretory stage were labelled as well as multivesicular dense bodies. Very little labelling was encountered in the ameloblasts in the transitional stage, where apoptotic bodies were clearly visible. The maturation stage also exhibited labelling of the secretory-like granules in the distal surface. The presence of gold particles over the plasma membrane is an indication that IGF-I receptor is a membrane-bound receptor. Furthermore, the intracellular distribution of the label over the endoplasmic reticulum supports the local synthesis of the IGF-I receptor. The absence of labelling over the transitional ameloblasts suggests that the transitional stage may require the non-expression of IGF-I as a prerequiste or even a trigger for apoptosis.

Ultrastructure of cementogenesis as affected by growth hormone in the molar periodontium of the hypophysectomized rat.

To document the effect of hypophysectomy and growth hormone replacement on the ultrastructure of cementogenesis in the developing rat third molar, 12 female Wistar rats were randomly allocated to normal control, hypophysectomized or hypophysectomized plus human growth hormone (for 10 days) treatment groups. The results of this study by electron and light microscopy and morphometry have shown that qualitative and quantitative changes occur in the organelles of cementoblasts forming cellular cementum as a result of hypophysectomy and growth hormone replacement. After hypophysectomy, the changes of less prominent nucleoli and nuclear pores, less prominent Golgi apparatuses and decreased endoplasmic reticulum can be interpreted as diminished cementum matrix biosynthesis--an interpretation that can be confirmed morphometrically by less cellular cementum formation. Growth hormone replacement for 10 days reactivates protein synthesis and cementogenesis as evidenced by ultrastructural changes in cementoblasts and a greater production of cementum.

The effect of chronic fluoride administration on rat condylar cartilage.

Morphometrical and histochemical techniques were used to demonstrate changes to the cartilage layer of the rat temporomandibular joint condyle following chronic exposure to fluoride. An increase in thickness of the cartilage layer was noted in rats given 100 parts per million sodium fluoride in drinking water. No significant changes were observed with either control or low dose (10 parts per million) groups. The observed thickening was attributable to an increase in number and size of cells of the lower hypertrophic zone. Accumulations of glycogen were observed in these cells, which reflects the inhibitory effect of fluoride on glycolysis. Stimulation of chondrocytes by fluoride may have delayed the normal processes of capillary invasion, resulting in thickening of the cartilage layer. No changes to staining patterns of immature or mature types of collagen were observed, nor did the staining pattern of detectable glycosaminoglycans change due to fluoride.

A histochemical study of the effects of high doses of sodium fluoride on dipeptidyl peptidase II activity in the rat incisor ameloblast.

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.

Microwave irradiation as a form of fixation for dental pulp.

This study investigated developments in microwave energy fixation and the general applicability of microwave fixation to studies of dental pulp. Rat mandibles with incisors were dissected out and immersed in various solutions before and after exposure to microwave energy. Histological examination showed no combination with microwave fixation to be equal in quality to control tissues fixed in formalin. Inferior but satisfactory results could be achieved by immersion of tissues in formalin after microwave irradiation, a step thought to protect the microwave fixed tissue from subsequent damage caused by decalcification. Good fixation of tissues outside the pulp was achieved using microwave energy. Further investigation is required to perfect microwave fixation of pulpal tissue.

Effects of acute doses of sodium fluoride on the morphology and the detectable calcium associated with secretory ameloblasts in rat incisors.

Fluoride in high concentrations is known to have an adverse effect on the formation of enamel. The effect of a single injection of two concentrations of sodium fluoride on inner enamel secretory ameloblasts was investigated morphologically by electron microscopy and functionally by assessing the location and relative amount of available calcium, using the potassium pyroantimonate method. The results showed that acute doses of fluoride interfere with the normal function of secretory ameloblasts. The increase in the population of lysosome-like structures observed after fluoride administration is suggestive of defects in the synthetic pathway. Concomitant with the effect of fluoride on secretory ameloblasts is an inhibition of enamel formation, resulting in incomplete enamel rods and leaving large remnants of Tomes' processes buried in the enamel. The distribution of the calcium pyroantimonate deposits found tends to support the concept of calcium traveling between the cells to the enamel. Acute doses of fluoride also reduce the amount of calcium available for complexing with pyroantimonate in the intercellular region.

Disturbances of tooth form and eruption in the microphthalmic (mi) mouse: a light and electron microscopic study.

Changes in the surrounding alveolar bone occur during tooth eruption. The microphthalmic (mi/mi) mouse suffers from osteopetrosis and lack of bone resorption; tooth form and eruption were examined in both affected mi/mi mice and unaffected litter-mates to determine the effect of osteopetrosis on tooth development and eruption. Paraffin sections of mandibles from 3, 7, 10, 13, 15 and 20-day-old mice were examined by light microscopy after staining with haematoxylin and eosin and for stable acid phosphatase activity. Mandibles from 15- and 20-day-old mice were examined by scanning electron microscopy. The ultrastructure of odontoblasts was observed in 15-day-old mice. Tooth eruption was significantly reduced in the mi/mi mice; the bone of affected mice increased in area with increasing age and marrow spaces narrowed. There was little bony remodeling in the mi/mi mouse, as indicated by layers of reversal lines. This lack of bone resorption affected tooth eruption and root formation. No abnormalities were detected in odontoblasts, suggesting functional normality, but the wide predentine layer in the mi/mi mouse may indicate an alteration in dentine mineralization.