Factors influencing dental arch form.

Dental arch length, bilateral intermolar distance, morphology of the anterior teeth, and bilateral intercanine distance have been reported as factors influencing the determination of dental arch form. However, studies evaluating the factors that influence the determination of the above-mentioned components are limited. Therefore, to verify these points, the present study aimed to examine factors influencing the determination of dental arch form using statistical methods.Data obtained from sample dental casts were analyzed using principal component and cluster analyses. By principal component analysis, 23 sets of information were summarized into three components for the maxilla and four for the mandible. As a result of cluster analysis using principal component scores, the maxillary and mandibular dental arches were classified into four forms, respectively.Dental arch length is an important indicator of dental arch size and is influenced by the bilateral interincisor distance of the maxilla and mandible, and the mesiodistal crown width of the incisors and premolars. In the mandible, canine width also affects dental arch length. Dental arch width also influences the determination of dental arch form. However, the distance between the anterior teeth and the distance between the molars are independent and have no effect on each other.

Histological Alterations from Condyle Repositioning with Functional Appliances in Rats.

OBJECTIVE: This study was designed to assess the morphological and histological alterations of the condyle of rats undergoing forward mandibular repositioning via functional appliance. MATERIALS AND METHODS: Functional appliances were mounted onto the upper jaws of rats. Morphological analysis was conducted on micro-CT images of sacrificed animals. Histological changes in condyle were examined by immunohistochemistry using proliferating cell nuclear antigen (PCNA), matrix metalloproteases (MMPs), vascular endothelial growth factor (VEGF), tissue inhibitors of matrix metalloproteinases (TIMP-1), interleukin 1b (IL-1ß), Aggrecan and Type II collagen. Osteoclast activity was identified by tartrate-resistant acid phosphatase (TRAP) staining. RESULTS: Morphological analysis confirmed the forward positioning of the condyles of rats by the appliance, but the position gradually returned to normal on days 14 after treatment. An increase in PCNA positive cells was observed in the posterior region of the condyles on days 7, whereas PCNA positive cells decreased in the anterior region. Aggrecan and Type II collagen localization increased in the posterior region throughout the entire period, but decreased in the anterior region on days 14. In both regions, IL-1ß and VEGF localization was significantly increased for 14 days while MMPs localization was evident throughout the entire period. The TRAP positive cells were significantly elevated on days 3 and 7. CONCLUSIONS: These results suggest that the functional appliance therapy induces significant morphological and histological changes in the anterior and posterior regions of the condyle and subsequently causes adaptive cellular functions such as chondrocyte differentiation and cartilage matrix formation.

Taste preference changes throughout different life stages in male rats.

Taste preference, a key component of food choice, changes with aging. However, it remains unclear how this occurs. To determine differences in taste preference between rats in different life stages, we examined the consumption of taste solutions and water using a two-bottle test. Male Sprague-Dawley rats of different ages were used: juvenile (3-6 weeks), young adult (8-11 weeks), adult (17-20 weeks), middle-aged (34-37 weeks), and old-aged (69-72 weeks). The intakes of the high and low concentration solutions presented simultaneously were measured. We observed that the old-aged group had lower preference ratios for 0.3 M sucrose and 0.1 M MSG in comparison with other groups. The preference ratio for 0.03 mM QHCl was higher in the middle-aged group than in the three younger groups and higher in the old-aged group than the juvenile group. The taste preferences for HCl and NaCl did not significantly differ among the age groups. The old-aged group tended to prefer high concentrations of sucrose, QHCl, NaCl, and MSG to low concentrations, indicating age-related decline in taste sensitivity. We also aimed to investigate differences between life stages in the electrophysiological responses of the chorda tympani nerve, one of the peripheral gustatory nerves, to taste stimuli. The electrophysiological recordings showed that aging did not alter the function of the chorda tympani nerve. This study showed that aging induced alterations in taste preference. It is likely that these alterations are a result of functional changes in other peripheral taste nerves, the gastrointestinal system, or the central nervous system.

A histological study of mineralised tissue formation around implants with 3D culture of HMS0014 cells in Cellmatrix Type I-A collagen gel scaffold in vitro.

We cultured HMS0014 Yub621b cells within a 3D collagen gel scaffold (Cellmatrix Type I-A) and aimed to study the fate and contribution of human bone-derived mesenchymal stem cells (MSCs) in the guided bone regeneration(GBR)-engineered tissue which has developed around the titanium (Ti) test dental implant (IP) in vitro. The light microscopy (LM) and transmission electron microscopy (TEM) results of the peri-IP tissue indicated that collagen fibrils of the Cellmatrix Type I-A gel were accumulated and fabricated to provide a 3D meshwork for proliferation and differentiation of the HMS0014 cells in the top (cell) layer; mineralisation of the GBR tissue had commenced since day 1 and became markedly deposited between days 7 and 14 of the experiment. TEM observation revealed sedimentation of cement line at the periphery of the interwoven Cellmatrix fibres and fibrils in the ECM scaffold of the GBR tissue; matrix vesicle-mediated and appositional collagen-mediated mineralisation were identified in the peri-IP ECM scaffold. The fine structure study of the plurimorphic osteoblast(Ob)-like osteogeneic cells demonstrated numerous membranous organelles related with vesicular trafficking, secretion and endocytosis in the cytoplasm; well-developed cytoskeleton networks and intercellular junctional complexes were also observed. The specimens on fluorescence immunohistochemistry (IHC) by confocal laser-scanning microscopy (LSM) showed the expression of LC3 and Cx43 associated with autophagic-lysosomal degeneration pathway and signal conduction mediated with gap junctions (GJS) in maintaining tissue homeostasis of the Ob-like cells which grew and degenerated in the 3D scaffold. Results from this in vitro study suggest that Ob-like HMS0014 cells actively regulate turnover of the peri-IP ECM to recapitulate the development and formation of osteoid tissue-engineered material which might contribute to augment osseointegration around the dental implant.

A preliminary study of the dental implant therapy--initial osteogenesis of human mesenchymal stem (HMS0014) cells on titanium discs with different surface modifications.

HMS0014 cells were GBR-engineered to proliferate and differentiate into mature osteoblast(Ob)-like cells, which initiated hard tissue matrix deposition in both monolayer and PuraMatrix 3-D cultures. Subsequently, the osteogenesis initiated with attachment/adhesion of HMS0014 cells on either Titanium (Ti) or Ti alloy discs modified with osteoconductive/ osteoinductive surface textures/substrates (e.g., Disc-AO, Disc-HA, Disc-SPI) was histologically assessed. The results obtained were as follows: 1) The HMS0014 cells actively proliferated and differentiated into mature Obs to initiate mineralisation of the ECM since day 1 in both monolayer and 3-D cultures; mineralization was prominently progressed between day 7 and day 14 of cultures. 2) The SEM of 60-minute(min)s specimens demonstrated a loose distribution of proliferating spherical-to-polygonal (10 to 40 microm in diameter, avg.) cells with a bulging cell body sending out many minute filopodia and some lamellipodia to attach with the substrate in the concavities. 3) In the 180-min specimens, the cultured HMS0014 cells actively proliferated and spread into flat, large polygonal cells with prominent lamellipodia and dendritic filopodia (30 microm x 90 microm to 100 microm x 200 microm, approx.) to employ cell-to-substrate and intercellular attachments. 4) On the other hand, the present immunohistochemistry of the attached HMS0014 cells demonstrated the co-expression of F-actin (actin filaments of the cytoskeleton) and CD51 (aV integrin) in both the 60-min and 180-min specimens. We concluded that the present GBR method enhanced HMS0014 cells to initiate an osteogenesis process with a direct bone-to-substratum contact on Ti discs which were subject to different surface modifications.

Isolated rat dental pulp cell culture and transplantation with an alginate scaffold.

Many studies have been conducted on tissue stem cells in the field of regenerative medicine, and cultured dental pulp mesenchymal cells have been reported to secrete dentin matrix. In the present study we used alginate as a scaffold to transplant subcultured rat dental-pulp-derived cells subcutaneously into the back of nude mice. We found that when beta-glycerophosphate was added to the culture medium, the mRNA of the dentin sialophosphoprotein (DSPP) gene coding dentin sialoprotein (DSP) and dentin phosphoprotein (DPP) was expressed, and an increase in alkaline phosphatase, an early marker of odontoblast differentiation, was also demonstrated. Six weeks after implantation, subcutaneous formation of radiopaque calcified bodies was observed in situ. Immunohistochemical and fine structure studies identified expression of type I collagen, type III collagen, and DSP in the mineralizing transplants, and isolated odontoblast-like cells began to form dentin-like hard tissue formation. Scattered autolyzing apoptotic cells were also observed in the transplants. The study showed that subcultured rat dental-pulp-derived cells actively differentiate into odontoblast-like cells and induce calcification in an alginate scaffold.

Human dental pulp cell culture and cell transplantation with an alginate scaffold.

Many studies on tissue stem cells have been conducted in the field of regenerative medicine, and some studies have indicated that cultured dental pulp mesenchymal cells secrete dentin matrix. In the present study we used alginate as a scaffold to transplant subcultured human dental pulp cells subcutaneously into the backs of nude mice. We found that when beta-glycerophosphate was added to the culture medium, dentin sialophosphoprotein mRNA coding dentin sialoprotein (DSP) was expressed. An increase in alkaline phosphatase, which is an early marker for odontoblast differentiation, was also demonstrated. At 6 weeks after implantation the subcutaneous formation of radio-opaque calcified bodies was observed in situ. Immunohistochemical and fine structure studies identified expression of type I collagen, type III collagen, and DSP in the mineralizing transplants. Isolated odontoblast-like cells initiated dentin-like hard tissue formation and scattered autolyzing apoptotic cells were also observed in the transplants. The study showed that subcultured dental pulp cells actively differentiate into odontoblast-like cells and induce calcification in an alginate scaffold.

Effects of peripheral inflammation on activation of ERK in the rostral ventromedial medulla.

In the present study, the activation of extracellular signal-regulated kinase (ERK) in the rostral ventromedial medulla (RVM) following the injection of complete Freund's adjuvant (CFA) into the rat hindpaw was examined in order to clarify the mechanisms underlying the dynamic changes in the descending pain modulatory system after peripheral inflammation. Phospho-extracellular signal-regulated kinase-immunoreactive (p-ERK-IR) neurons were observed in the nucleus raphe magnus (NRM) and nucleus reticularis gigantocellularis pars alpha (GiA). Inflammation induced the activation of ERK in the RVM, with a peak at 7 h after the injection of CFA into the hindpaw and a duration of 24 h. In the RVM, the number of p-ERK-IR neurons per section in rats killed at 7 h after CFA injection (14.2 +/- 1.7) was significantly higher than that in the control group (4.5 +/- 0.9) [P < 0.01]. At 7 h after CFA injection, about 60% of p-ERK-IR neurons in the RVM were serotonergic neurons. The percentage of RVM serotonergic neurons that are also p-ERK positive in the rats with inflammation (20.5% +/- 2.3%) was seven times higher than that in control rats (2.7% +/- 1.4%) [P < 0.01]. These findings suggest that inflammation-induced activation of ERK in the RVM may be involved in the plasticity in the descending pain modulatory system following inflammation.

Morphological classification of mandibular dental arch forms by correlation and principal component analyses.

To evaluate the morphology of dental arches, 53 (male: 29, female: 24) paired casts having normal dentitions and occlusion were selected from 396 (age: 18 to 26 years old; male: 257, female: 139) sets of dental study models. The mandibular dentitions were preliminarily classified as square, round-square, round and round V-shaped arches based on the conventional morphological descriptions. Midpoints of the incisor edge (I1R, I1L, I2R, & I2L), summits of the cuspids (CR & CL), buccal cusps of the premolars (P1R, P1L, P2R, & P2L), mesial buccal cusps of the first and second molars (M1R, M1L, M2R, & M2L), and the midpoint (A) of line I1R-I1L were designated as reference points. From A, let a vertical line intersected line M2R-M2L at reference point B. The line A-B intersected CR-CL at reference point E. We evaluated 1) the protrusion of the cuspids by 1. angle I2R-CR-P1R (angle R) + angle I2L-CL-P1L (angle L); 2) the curvature of the anterior teeth by 2. (A-B)/(CR-CL), 3. (180 degrees-angle(CR-A-CL), and 4. (A-E)/(CR-CL); 3) the length to width ratio of the dental arch by 5. (A-B)/(M2R-M2L); 4) the degree of roundness of the mandibular arch by estimation of 6. (rtheta5 - rtheta4)R + (rtheta5 - rtheta4)L; and 5) an item 7. for the differentiation of type I and type II round-square arches by relating the bilateral contour and position of break line P1-P2-M1-M2 (i) to line P1-M2 (ii). The data of items 1., 2., 3., 4., 5., and 6. were further standardized and summarized into three essential principal components: 1) the curvature of the anterior teeth, 2) the curvilinear contour of the dental arch, and 3) the length-to-width ratio of the dental arch. The results indicated that: 1) 36 cases (67.9%) of the mandibular dentitions were round-square arches which showed no prominent principal component. 11 cases (20.8%) were square arches and 6 cases (11.3%) were round V-shaped arches; no round arches was found in mandibular dentitions. 2) Statistical analysis indicated significant differences of items 3., 4. and 6. in various mandibular arches (Student's t-test). 3) By examination of the three principal components, significant differences of item 5. between the round V-shaped arches and square and round-square mandibular arches were evident (Student's t-test). The present study elucidated that morphology of the mandibular arch was determined by a parameters representing the curvature of anterior teeth (composed of items 2., 3. and 4., and another parameter (item 6.) representing roundness of the mandibular arch.

Increase of galanin-like immunoreactivity in rat hypothalamic arcuate neurons after peripheral nerve injury.

Galanin and galanin receptors are widely distributed within the central nervous system, and may play important roles in pain signaling and modulation. In the present study, we examined the galanin immunoreactivity (IR) in the hypothalamus and the amygdala following peripheral nerve injury. Four weeks after the operation, the ipsilateral mechanical threshold in the spared nerve injury (SNI) group (0.87 +/- 0.33 g) was significantly lower than that in the sham group (12.53 +/- 3.41 g; P < 0.05). In the SNI group, the number of galanin-IR neurons per section in the arcuate nucleus (Arc) of the hypothalamus was 10.2 +/- 1.7, significantly higher than that in the sham group (5.6 +/- 1.0; P < 0.05). These data suggest that the galanin-ergic neurons in the Arc may be involved in the functional modulation of descending pain modulation system following peripheral nerve injury.

Blockade of peripheral 5HT3 receptor attenuates the formalin-induced nocifensive behavior in persistent temporomandibular joint inflammation of rat.

The role of peripheral 5HT3 receptors in the orofacial nocifensive behavior induced by the injection of formalin into masseter muscle was evaluated. The behavioral activities evoked by the formalin injection exhibited a biphasic response in the rats with or without temporomandibular joint (TMJ) inflammation (CFA group or non-CFA group). The orofacial nocifensive behavioral activity was enhanced after TMJ inflammation. Systemic administration of tropisetron, 5HT3 receptor antagonist, reduced the nocifensive behavioral activities in the late phase of orofacial formalin test in CFA group, but not in non-CFA group. Local administration of tropisetron into the masseter muscle in CFA group, but not in non-CFA group also attenuated the behavioral activities in the late phase. Unexpectedly, low dose of local tropisetron reduced the nocifensive behavioral activities in the early phase of orofacial formalin test in CFA group. These data suggest that induction of TMJ inflammation causes the elevation of the orofacial nocifensive behavioral activities evoked by formalin injection into masseter muscle, and that peripheral 5HT3 receptors may play a critical role in nociception and the transmission of orofacial pain.

A morphological study on the classification of maxillary dental arches.

To evaluate the morphology of dental arches, 62 (male: 36, female: 26) paired casts having normal dentitions and occlusion were selected from 396 (age: 18 to 26 years old; male: 257, female: 139) sets of dental study models. The maxillary dentitions were preliminarily classified as square, round-square, round and round V-shaped arches based on the conventional morphological descriptions. Midpoints of the incisor edge (I1(R), I1(L), I2(R), & I2(L), summits of the cuspids (CR & CL), buccal cusps of the premolars (P1(R), P1(L), P2(R), & P2L), mesial buccal cusps of the first and second molars (M1(R), M1(L), M2(R), & M2(L)), and the midpoint (A) of line I1(R)-I1(L) were designated as reference points. From A, let a vertical line intersected line M2(R)-M2(L) at reference point B. The line A-B intersected C(R)-C(L) at reference point E. We evaluated 1) the protrusion of the cuspids by (1) angle I2(R)-C(R)-P1(R) (angle R) + angle I2(L)-C(L)-P1(L) (angle L); 2) the curvature of the anterior teeth by (2) A-B/C(R)-C(L), (3) 180 degrees - angle C(R)-A-C(L), and (4) A-E/C(R)-C(L); 3) the length to width ratio of the dental arch by (5) A-B/M2(R)-M2(L); 4) the degree of roundness of the maxillary arch by estimation of (6) (rtheta5 - rtheta4)R + (rtheta5 - rtheta4)L; and 5) an item (7) for the differentiation of type I and type II round-square arches by relating the bilateral contour and position of break line P1-P2-M1-M2 (i) to line P1-M2 (ii). The data of items (1), (2), (3), (4), (5), and (6) were further standardized and summarized into three essential principal components: 1) the curvature of the anterior teeth, 2) the curvilinear contour of the dental arch, and 3) the length-to-width ratio of the dental arch. The results indicated that: 1) 60% of the maxillary dentitions were round-square arches which showed no prominent principal component; 2) square maxillary arches distinctly showed a small (1) angle R + angle L; 3) round arches were characteristic by small (6) (rtheta5 - rtheta4)R + (rtheta5 - rtheta4)L values; and 4) round V-shaped arches had large (1), (3) and (4) values.

Immunohistochemical study of genesis of the mouse oral vestibule.

Immunohistochemical light microscopy by the enhanced polymer one-step staining (EPOS) method and immunohistochemical confocal laser scanning microscopy by the labelled streptavidin biotin (LsAB) method were performed on the developing vestibular lamina (VL) of fetal mice at stages from E11 to E14, and the immunohistochemical findings were compared with the findings in the developing dental lamina (DL) and tooth germ. pRb and PCNA were immunolocalized and found to be related to the temporal and spatial expression of cytokines and receptors in the primary epithelial band, VL, DL/enamel organ, and associated mesenchyme. The following results were obtained: 1) EGF, TGFalpha, EGFR, PCNA, FGF2, pRb, and FGFR1-4 were immunolocalized in the developing tissues. 2) Cytokine expression patterns indicated that the EGF family and FGF2 essentially induced VL generation and cell proliferation. 3) FGFR1 was diffusely localized in the primary epithelial band, but was strongly expressed in the E12-14 VL and DL/enamel organ. By contrast, EGFR internalization was observed in the differentiating E13 VL. 4) Expression of pRb was intensely localized in the stratum germinativum of the E13 VL and corresponded to CK-10 expression in the keratinizing VL. The results of this study suggest a mechanism in which FGFR1 regulates pRb to induce proliferation of cells in the VL and DL/enamel organ, and, in particular, to incite keratocyte differentiation and subsequent exfoliation of keratinizing VL cells.