Rat Molar Tooth Research Articles

Effect of voxel size on accuracy of cone beam computed tomography–aided assessment of periodontal furcation involvement

To assess the accuracy of cone beam computed tomography (CBCT) images obtained at different voxel sizes in detecting ligature-induced furcation involvement. Furcation lesions were prepared in 36 molar teeth in rats, and 24 intact teeth served as controls. CBCT Kodak 9000 images at three different voxel sizes were acquired: 0.076 mm, 0.100 mm, and 0.200 mm. Four observers assessed 3 sets of images using a 5-point evaluation scale. κ-Coefficients were calculated for intra- and interobserver agreement. Receiver operating characteristic analysis measured the true positive rate (TPR), true negative rate (TNR), and area under the curve (AUC). Accuracy values were compared by using Mood's Median Chi-Square (α<0.05). Intra- and interobserver agreement ranged from 0.600 to 0.999 and from 0.366 to 0.573, respectively. Highest median AUC and true positive rate (TPR) values were obtained for voxel size of 0.076 mm. The highest median TNR values were obtained for the voxel size 0.1 mm. There were no significant differences (P≥.05) among the median AUC, TPR, or TNR values for the 3 different CBCT voxel sizes. Given the limitations of this study, all voxel sizes performed similarly in the detection of furcation involvement.

Non-autologous endodontic pulp regeneration approach in molar teeth of the rat

The dental pulp is an important soft connective tissue which is able to produce dentin over time as a reaction on external stimuli. It also maintains the biological and physiological vitality of the dentin. Due to this the pulp is essential for teeth homeostasis. However, dental caries is still one of the most prevalent health problems in dentistry and therefore, one major cause for early loss of the dental pulp vitality and subsequent tooth extractions. Meanwhile the potential for successful pulp regeneration therapy is increasing due to advances in the field of regenerative endodontics. Thus, adequate experimental animal models are required for testing and validating these new regenerative therapies. Rodents and rats in particular, are relevant models for experimental periodontal research. The breeding and housing costs of rats are relatively low facilitating studies with sufficient numbers for statistical analysis in comparison to bigger sized mammals like beagle dogs, miniature pigs or monkeys. Additionally, rat molar teeth and pulps are characterized by similar anatomical, histological, biological and physiological features to human teeth. Essential biological reactions of the pulp tissue and the interaction during the different stages of wound healing of rat molar teeth are comparable to that of other mammals. However, despite of the multiple research activities in the field of regenerative endodontics and the above mentioned advantages of the rat model only rare in vivo studies are published. Therefore, the presented study aimed to introduce the rat molar teeth as a valid model for studying dental pulp stem cell based endodontic tissue regeneration. Human dental pulp stem cells were implanted into the pulp of immunodeficient rats (RNU rats). Cell growth was supported by a collagenous membrane, which was applied on top of the cells after implantation. After closing the pulpal cavity with a light-polymerisable resin human dental pulp stem cells were able to maintain cell viability in the rat molar pulp niche for at least three weeks. This demonstrated the suitability of immunodeficient RNU rats for non-autologous dental stem cell based endodontic tissue engineering approaches.

Decreased face primary motor cortex (face-M1) excitability induced by noxious stimulation of the rat molar tooth pulp is dependent on the functional integrity of face-M1 astrocytes.

Acute inflammatory dental pain is a prevalent condition often associated with limited jaw movements. Mustard oil (MO, a small-fiber excitant/inflammatory irritant) application to the rat molar tooth pulp induces increased excitability (i.e., central sensitization) of trigeminal medullary dorsal horn (MDH) nociceptive neurons that can be modulated by MDH application of the astrocytic inhibitor methionine sulfoximine (MSO). The objectives of the study were to determine whether MO application to the rat right maxillary first molar tooth pulp affects left face-M1 excitability manifested as altered intracortical microstimulation thresholds for evoking electromyographic activity in the right anterior digastric (RAD, jaw-opening muscle), and whether MSO application to face-M1 can modulate this MO effect. Under Ketamine general anesthesia, Sprague-Dawley male rats had a microelectrode positioned at a low-threshold (≤30 μA) face-M1 site. Then MO (n = 16) or control solution (n = 16) was applied to the previously exposed tooth pulp, and RAD threshold was monitored for 15 min. MSO (0.1 mM, n = 8) or saline (n = 8) was then applied to the face-M1, and RAD thresholds were monitored every 15 min for 120 min. ANOVA followed by post hoc Bonferroni was used to analyze data (p < 0.05). Within 15 min of MO (but not control) pulp application, RAD thresholds increased significantly (p < 0.001) as compared to baseline. One hour following MSO (but not saline) application to the face-M1, RAD thresholds decreased significantly (p = 0.005) toward baseline. These novel findings suggest that acute inflammatory dental pain is associated with decreased face-M1 excitability that may be dependent on the functional integrity of face-M1 astrocytes and related to mechanisms underlying limited jaw movements in acute orofacial pain conditions.

Promotion of Bone Formation around Alendronate-immobilized Screw-Type Titanium Implants after Implantation into Rat Molar Tooth Sockets

Promotion of Bone Formation around Alendronate-immobilized Screw-Type Titanium Implants after Implantation into Rat Molar Tooth Sockets

Immunocytochemical and biochemical detection of the urokinase-type plasminogen activator receptor (uPAR) in the rat tooth germ and in lipid rafts of PMA-stimulated dental epithelial cells

Urokinase-type plasminogen activator receptor (uPAR) regulates pericellular proteolysis by binding the serine proteinase urokinase-type plasminogen activator (uPA) that promotes cell surface activating of plasminogen to plasmin. In addition, uPAR as a glycosylphosphatidylinositol (GPI)-anchored signaling receptor affects cell migration, differentiation, and proliferation. The aim of the present study was to monitor the occurrence and distribution pattern of uPAR in cells of the rat molar tooth germ. By means of immunocytochemistry moderate, uPAR immunoreactivity was detected in epithelial cells of the enamel organ and in ameloblasts and odontoblasts. RT-PCR and Western blotting experiments demonstrated the expression of uPAR in phorbol 12-myristate 13-acetate (PMA)-stimulated dental epithelial cells (HAT-7 cells). A substantial part of uPAR was detected in the detergent-insoluble caveolin-1-containing low-density raft membrane fraction of HAT-7 cells suggesting a partial localization within lipid rafts. However, co-immunoprecipitation experiments showed that uPAR and caveolin-1 do not associate with each other directly. Cell stimulation experiments with PMA indicated that protein kinase C (PKC)-mediated signaling pathways contribute to the expression of uPAR in cells of the enamel organ. The localization of uPAR in membrane rafts provides a basis for further investigations on the role of uPAR-mediated signaling cascades in ameloblasts.

Physical and Morphological Studies of the Susceptible Fissure Enamel against Dental Caries in Rat Molar Teeth

Physical and Morphological Studies of the Susceptible Fissure Enamel against Dental Caries in Rat Molar Teeth

Fine Structure of Susceptible Enamel Lamellae against Dentin Caries in the Fissures of Rat Molar Teeth

Fine Structure of Susceptible Enamel Lamellae against Dentin Caries in the Fissures of Rat Molar Teeth

In situ study of the gelatinase activity in demineralized dentin from rat molar teeth

Matrix metalloproteinases (MMPs) in dentin are believed to participate in various physiological and pathological events in coronal dentin, but their exact source and location is not clear. The purpose of this study was to evaluate the activity of gelatinases in decalcified rat molars crowns by in situ zymography. Hemi-mandibles of five male Wistar rats were fixed in paraformaldehyde, decalcified in EDTA and glycerol solution and embedded in paraffin. Sections from the region of molar teeth were incubated with or without DQ gelatin in 50mM Tris–CaCl2 at 37°C for 2h and observed by means of confocal microscopy. Gelatinolytic activity was observed throughout the coronal dentin with varying intensities in different locations. High gelatinase activity was observed in the dentinal tubules, dentin–enamel junction (DEJ) and predentin, and it was weaker and less uniform in the intertubular dentin. This study shows that the location of gelatinase and relative activity can be detected by means of in situ zymography and confocal microcopy, and this methodology may provide a useful tool in studies on the role of gelatinases in tooth development, maturation and in pathological conditions.

Influence of aging on experimental gastrointestinal motility in extraction of rat molar teeth

The purpose of the present study was to determine whether different consistency of diet and malocclusion induced by the extraction of molar teeth on the masticatory organs modulated gastric acid secretion, gastric emptying and intestinal transit in young and elder rats. Male Wistar rats (young, 5 weeks; elder, 1.5 years) were used in this experiment, and were divided into 2 groups. Group one (G1) was maintained with solid diet, Group two (G2) with mud diet. Further, the mandibular molar teeth of G2 were extracted. The experimental period was 10 weeks. The effect of aging and malocclusion on the parameters of gastric secretion was examined using pylorus-ligated rats. The gastric emptying rate (GER) and small intestinal transit rate was determined in rats by evans blue from the stomach and charcoal from the small intestinal, respectively. In pylorus-ligated rats, Young-G2 rat of gastric juice volume, acid output and pepsin secretion remarkably showed significant decrease in comparison with Young-G1 group, but there was not significant difference between Elder-G1 and Elder-G2. GER of Young-G2 rat group was 44.2±7.9%, significantly lower than that of Young-G1 rat group (61.6±8.8%, P <0.01), but Elder-G1 rat group were not significantly different than those of rats of Elder-G2 group. In small intestinal transit rates of charcoal meals, G1 and G2 of young were 73.3±9.1 and 55.1±8.6%, respectively, and Elder groups were 61.7±9.8 and 52.6±7.7%, suggesting an insignificant effect on diet. These results suggest that the diet and malocclusion, induced by extraction of mandibular molar teeth of young rat groups, may have a great influence compared to elder rat groups.

Histopathological and radiographic evaluation of rat molar teeth after traumatic injury-a pilot study

Objective:Radiographic and histopathological evaluation of mechanical trauma on the development of maxillary molars in rat.Materials and Methods:Experimental study with 7-day-old five new born Wistar rats. Trauma induced on 7th postnatal day with #55 K file. Right first molar region of the maxilla was the experimental side, and the left side as control. Each animal was sacrificed for histopathological observation on 8th day (immediately next day of injury), 10th day (after 3 days of injury), 13th day (Enamel completion), 19th day (Emergence into oral cavity) and 42nd day when all the teeth came into functional occlusion.Results:On the 42nd day experimental side of the study, the 1st molar tooth had not erupted and the radiographs taken revealed conglomerate mass suggestive of complex odontome. Reduced tooth size, altered enamel and dentin formation, abscess formation and complex odontome like structure were observed histopathologically.Conclusion:The diversity of morphlogical and histopathological changes to intrusive force implies intricate mechanism involved in tooth development.

Influence of aging on experimental gastrointestinal motility in extraction of rat molar teeth

Influence of aging on experimental gastrointestinal motility in extraction of rat molar teeth

Tooth movements are guided by specific contact areas between the tooth root and the jaw bone: A dynamic 3D microCT study of the rat molar

Teeth sustain high loads over a lifetime and yet intact tooth failure is rare. The different structures of the tooth, jaw bone and the intervening soft periodontal ligament enable the tooth to endure repeated loading during mastication. Although mechanical and functional properties of the different components are thoroughly investigated, the manner in which the whole tooth functions under load is still enigmatic. A custom-made loading system inside a microCT scanner was used to directly visualize the root movements in relation to the jaw bone as the rat molar tooth was loaded. At low loads no contact was observed between the root surface and the bone, whereas at higher loads three specific contact areas between the root surface and the jaw bone were observed. These contact areas restrict tooth movement in the buccal-lingual direction, but enable the tooth to rock in a "seesaw" like manner in the distal-mesial direction. The contact areas appear to play a role in determining tooth motion and in turn define the manner in which the whole tooth moves when loaded. These observations are important for understanding basic structure-function relations of the tooth-PDL-bone system, and have direct implications for better understanding pathological and therapeutic processes in orthodontics, periodontics and jaw bone regeneration.

Characterization of Rat Apical Tissues in Different Root Development Stage

In this study, we try to compare the histological characteristics and the odontogenic capability of apical tissues (AT) at different root development stages of rat molar teeth. AT of mandibular first molars from 8-day-old, 21-day-old, and 35-day-old Sprague-Dawley rats were selected as being representative of root-initiating, root-forming, and root-completing stages, respectively. Cell counting, flow cytometry assays, alkaline phosphatase activity, alizarin red staining, and reverse transcription polymerase chain reaction were performed to assess the proliferation and mineralization potential of apical tissue cells at different stages of root development in vitro. In vivo transplantation of apical tissue cells combined with ceramic bovine bone was used to characterize the differentiation capacity. It was shown that there was a structurally and functionally dynamic change in the apical tissue of developing tooth root of rats, of which the unique developmental potential will reduce gradually with the ending up of root development. The AT of root-initiating and root-forming stage exhibited much higher proliferation and tissue-regenerative capacity than those of root-completing stage. Our present results indicate that the apical tissue, with the sustainable developmental ability throughout almost the whole process of tooth development, can yet be regarded as a competent candidate source for root/periodontal tissues regeneration.

Rat molar teeth as a study model for direct pulp capping research in dentistry

The aim of this review is to evaluate the suitability of rat molar teeth in preclinical evaluation of medical devices for direct pulp capping. The ISO standard 7405 states clearly that only non-rodent mammals are suitable species for animal research in dentistry. Furthermore, without clear justification a considerable number of researchers previously rejected results of animal experiments concerning preclinical evaluation of the biocompatibility of dental materials undertaken in rat molar teeth. However, in the past 50 years about 70 studies have been published using rat molar teeth in order to evaluate direct pulp capping, pulpotomies and tissue reactions after pulp exposure. Numerous studies showed that the healing of rat molar pulp tissue after direct pulp capping is histologically comparable with humans and other animal species pulp tissue. Rat molar teeth, including pulp tissue, can be seen anatomically, histologically, biologically, and physiologically as miniature human molar teeth. Hence, the essential biological reactions of the pulp tissue and the interaction during the different stages of wound healing of rat molar teeth are comparable with that of other mammals. Rat molar teeth are a valid study model in order to provide valuable data concerning pulp tissue reaction after direct pulp capping and related questions in dentistry. Therefore, the use of rats may significantly reduce the number of currently used higher animals in research. Tests in higher developed animals should be limited to experiments which clarify inconsistent results. However, some technical difficulties, like the small size of rat molar teeth must be dealt with before undertaking any research.

Transforming growth factor beta inducible apoptotic cascade in epithelial cells during rat molar tooth eruptions

In tooth eruptions, the presence of apoptotic epithelial cells at the eruption site has been reported, but the factors that induce apoptosis in these cells remain to be elucidated, as do the induction pathways. In this study, we focused our attention on transforming growth factor beta (TGF-beta), which is known to induce apoptosis during embryonic development. Oral epithelium and dental lamina of maxillary first molars in 8- and 15-day-old rats were used to investigate the induction pathway of apoptosis by performing the immunohistochemical tests outlined below and assessing the characteristics of cells that undergo apoptosis by transmission electron microscopy in rats 8 and 15 days after birth. We examined TGF-beta-receptor 1, TGF-beta inducible transcription factor 1 (TIEG1), NADPHoxidase 4 (Nox4), cytochrome c, caspase-3 (active form and pro-enzyme), apoptosis-inducing protein Daxx, apoptosis signal-regulating kinase 1 (ASK1), glycogen synthase kinase-3 beta phosphorylated on serine 9 (p-GSK-3beta), and beta-catenin. We also performed periodic acid Schiff (PAS) reaction and terminal deoxynucleotidyl transferase-mediated dUTD nick end labeling (TUNEL) staining. At eruption sites 8 days after birth, reactions to TGF-beta-receptor 1, TIEG1, Nox4, cytochrome c, caspase-3, p-GSK-3beta, and beta-catenin, and PAS-positive cells were observed in areas close to the basal layer of oral epithelium through to the center of the dental lamina, but no reaction to Daxx or ASK1 was noted at these sites. Electron microscopy revealed the accumulation of glycogen granules in the cells that showed reactions to the above-mentioned markers as well as in the spaces among them. In the rats 15 days after birth (immediately before tooth eruption), the PAS-positive cells that showed reactions to the above antibodies remained on the buccal side of the epithelium, and high-electron-density apoptotic bodies and TUNEL-positive bodies were noted. Therefore, during tooth eruption, TGF-beta may induce apoptosis of cells rich in glycogen granules, and cytochrome c and caspase-3 may function to induce apoptosis. In addition, reactive oxygen species may be involved in this induction pathway via TIEG1 and Nox4 without involvement of Daxx and ASK1. Moreover, overexpression of p-GSK-3beta and beta-catenin may also contribute to apoptosis of oral epithelium at the eruption site and dental lamina cells. Glycogen storage mediated by p-GSK-3beta and crosstalk between the TGF-beta and Wnt signaling pathways may participate in the formation of tooth eruption passage.

Morphometric and immunohistochemical investigation of tooth development in rats prenatally exposed to ethanol

The purpose of this study was to clarify the effect of ethanol on rat molar tooth development. Pregnant rats were fed 25%-ethanol solutions or ethanol-free water up to delivery, and tooth germs, as well as fully developed teeth, obtained from the offspring (ethanol-exposed rats or control rats) were subjected to morphometric analysis and immunohistochemistry. No significant differences were observed in terms of tooth size between the ethanol-exposed and control rats. Moreover, no abnormalities in cusp number, form or dental surface structure was noted in the ethanol-exposed rats. In the ethanol-exposed rats, the tooth development showed a slight retardation in comparison to the controls. Morphological alteration in tooth germs was not observed in the experimental rats. Both bone morphogenetic protein-4 and basic fibroblast growth factor-2 were immunolocalized in the cells composing the tooth germs in the ethanol-exposed rats. No differences, however, were found in immunostaining intensity between the ethanol-exposed and control rats. Taken together, the results indicate that the effect of ethanol on tooth development was weaker than expected in the rat FAS model.

Localization of Runx2, Osterix, and Osteopontin in Tooth Root Formation in Rat Molars

Cementogenesis starts with the differentiation of cementoblasts. Mature cementoblasts secrete cementum matrix. Cementum components are similar to bone; moreover, cementoblasts possess many characteristics similar to those of osteoblasts. Runx2 and osterix, the transcriptional factors for osteoblast differentiation, participate in tooth formation. However, the characteristics of Runx2 and osterix during the differentiation process of cementoblasts remain unclear. In this study, we examined the immunolocalization patterns of Runx2, osterix, and osteopontin during rat molar tooth formation. Periodontal ligament cells and osteoblasts located on the alveolar bone surface showed immunoreactivity for Runx2. Colocalization of Runx2 and osterix was detected in cementoblasts, which penetrated the ruptured Hertwig's epithelial root sheath and attached to root dentin. Moreover, osteopontin was observed in Runx2-positive cementoblasts facing the root surface. However, the cells adjacent to cementoblasts showed only Runx2 reactivity. Neither Runx2 nor osterix was seen in cementocytes. These results suggest that both Runx2 and osterix are important for differentiation into cementoblasts. Additionally, osterix may be indispensable for transcription of osteopontin expression.

Low‐dose doxycycline inhibits bone resorption associated with apical periodontitis

To test the effect of low-dose doxycycline on bone resorption associated with apical periodontitis. Apical periodontitis was induced by occlusal pulp exposure in the mandibular first molars of 36 rats. Animals were divided into three groups of 12: group A received doxycycline in drinking water at a dose of 5.85 mg day(-1); group B received a dose of 1.48 mg day(-1) (one-quarter of the original dose); and group C received no medicament and served as the control. A bioassay determined the doxycycline serum levels. After 21 days, the mandibles were removed, radiographed and the radiographs scanned to generate digital images. These images were analysed morphometrically and the total area of the periapical bone resorption of the mesial and distal roots of each tooth was determined and used to compare the groups. Statistical analysis was completed using anova with repeated measures. The mean doxycycline serum level in group A was 0.22(+/-0.03) microg mL(-1) and in group B below the detection level of the assay (<0.062 microg mL(-1)). The mean area of the periapical bone resorption in the control group C was 2.91(+/-0.61) mm(2). In animals treated with a low-dose doxycycline, the mean size of the bone resorption was significantly smaller at 1.59(+/-0.59) mm(2) (group A) and 1.72(+/-0.85) mm(2) (group B) (P = 0.001). No significant difference was found in the area of the bone resorption between these two groups A and B. Low-dose doxycycline reduced the area of bone resorption associated with apical periodontitis in the mandibular first molar teeth of rats.

Pulp and root development after partial extrusion in immature rat molars: a histopathological study.

Experimental studies were undertaken in immature rat molar teeth to investigate the relationship between root development and the prognosis after partial extrusion. The experimental partial extrusion was induced at 1/2 root formation, 2/3 root formation and 3/4 root formation, using an extrusion device designed by the author. Histopathological studies and SEM observation of the pulpal tissue in rat maxillary first molars with partially formed roots after partial extrusion were conducted and evaluated. The degree of extrusion was changed according to the development of the root to produce a similar luxated state. The rats were sacrificed immediately after experimental extrusion, immediately after operation, and after 3 days, 7 days, 14 days, 30 days, and 60 days to compare the histopathological and SEM findings between experimental groups and control groups. Immediately after the extrusive injury, ablations of the odontoblast layer from dentin were seen with retention of a fluid component in the ablated spaces in the pulp. On 1 and 3 days after surgery, cellular components such as erythrocytes and leukocytes due to bleeding were seen. Ablations of the odontoblast layer were seen in the coronal pulp most frequently in the pulp horn, cervical area and floor of the pulp chamber. In general, pulp reaction after extrusion in groups with more complete root formation was more severe than in groups with less complete root formation. It is suggested that the more complete the root formation, the greater the effect on the pulp as result of partial extrusion, while with less complete root formation, the greater the malformation of the root apices.

Rat wct mutation induces a hypo-mineralization form of amelogenesis imperfecta and cyst formation in molar teeth

Our previous findings have demonstrated that the rat autosomal-recessive mutation, whitish chalk-like teeth (wct), induces enamel defects resembling those of human amelogenesis imperfecta (AI) in continuously growing incisor teeth. The present study clarifies the effect of the wct mutation on the morphogenesis and calcification of rat molar teeth. Formalin-fixed maxillae obtained from animals aged 4-30 days were examined by electron probe micro-analysis (EPMA) and by immunocytochemistry for amelogenin, ameloblastin, and enamelin. There were no distinct differences in the calcium and phosphorous contents and the amount of enamel between homozygous mutant and wild-type teeth during postnatal days 4-11. Although the mineral density in the enamel matrix considerably increased in the wild-type teeth until day 15, no changes occurred in mutant teeth during days 11-30. The immunoreactivity for enamel proteins in the secretory-stage ameloblasts in mutant teeth was similar to that in the wild-type teeth, and subsequently mutant maturation-stage ameloblasts became detached from the enamel surface, resulting in odontogenic cyst formation between the enamel organ and matrix until day 7 and the expansion of the cyst around the whole tooth crown on day 15. On day 30, the erupted mutant teeth presented morphological changes such as enamel destruction and tertiary dentin formation in addition to low mineral density in the enamel. Thus, the wct mutation prevents mineral transport without disturbing the synthesis of enamel proteins in molar teeth because of the absence of maturation-stage ameloblasts, in addition to the occurrence of odontogenic cysts.