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Synovial hyperplasia is a feature of degenerative temporomandibular joint (TMJ) disease. However, the mechanism by which hyperplasia progresses in the TMJ is unknown. Based on the hypothesis that the oxidative stress generated by mechanical loading causes degenerative changes in the TMJ synovium, we investigated the generation of the highly reactive species, peroxynitrite, and the occurrence of DNA damage in the synovium. After condylar hypermobility of rat TMJs, a marker of peroxynitrite, nitrotyrosine, was localized to the nuclei and cytoplasm of the synovial lining cells and fibroblasts in synovitis-induced TMJ. DNA single-strand breaks were found in the nuclei of the synovial cells only after enzyme treatment, whereas DNA double-strand breaks were not detected. These findings indicate that condylar hypermovement induces the proliferation of synovial cells, and suggest that oxidative stress leads to the progression of synovial hyperplasia via DNA damage of the synovial cells in TMJs after mechanical loading. [ABSTRACT FROM AUTHOR]

Capsaicin, the pungent substance in hot peppers, evokes a sensation of burning pain by stimulating the vanilloid receptor 1 (VR1) on primary afferent neurons. Immunohistochemistry revealed that the taste papillae in the tongue and palate are richly innervated by VR1-immunoreactive nerve fibers. Furthermore, VR1 protein expression was seen in the epithelium facing the oral cavity, although taste cells seemed to be devoid of VR1. The most conspicuous VR1 expression was observed in the epithelial cells of the palatal rugae, although there were no VR1-immunoreactive nerves there. The finding that VR1 is expressed not only in primary afferents but also in oral epithelial cells suggests that it is of great importance in the perception of capsaicin, heat, and acid in the mouth. Since VR1 is known to play a key role in nociception and inflammatory pain, it may be a new target for the treatment of oral pain. [ABSTRACT FROM AUTHOR]

NF-κB plays a pivotal role in pathogenesis in general arthritis. However, the participation of NF-κB in inflammation of the temporomandibular joint (TMJ) is poorly understood. We examined NF-κB expression in rat TMJs with synovitis induced by condyle hypermobility. By immunohistochemistry, NF-κB immunoreactivity was found mainly in the cytoplasm, not the nucleus, of the synovial lining cells of inducedsynovitis and control TMJs. Southwestern histochemistry, a new method for detecting transcription factors, showed greater NF-κB expression in the nucleus of the synovial lining cells in the hypertrophic synovium than in control synovium. Increased numbers of the synovial lining cells with immunoreactivity for inducible nitric oxide synthase (iNOS), which is transcriptionally regulated by NF-κB, were also seen in the inflamed synovium. These findings indicate that excess mechanical stress increases NF-κB activation in the TMJ and suggest that active NF-κB is involved in the progression of TMJ inflammation. [ABSTRACT FROM AUTHOR]

Because the ultrastructure of the trigeminal sensory nerves in dentin, especially in relation to odontoblasts, remains to be clarified, we investigated the relationship between the trigeminal sensory nerves and the odontoblast processes using the anterograde axonal transport technique by injecting wheat germ agglutinin-horseradish peroxidase (WGA-HRP) into the rat trigeminal ganglion. Light microscopically, the nerves labeled with WGA-HRP were mainly concentrated at the pulpal horn, forming a nerve plexus at the subodontoblastic region and penetrating the predentin/dentin about 50 to 70 pm. Ultrastructurally, HRP reaction products were observed intra-axonally in the myelinated (Aδ) and unmyehnated (C) axons in the subodontoblastic region. Most nerves lost the Schwann sheath and were naked in the predentin/dentin. The labeled varicosities were close to the odontoblast processes in the dentinal tubules. No synaptic structures could be detected between the varicosities and the odontoblasts, but a gap about 20 nm wide was found between them. One type of varicosity was a rich mitochondria-containing varicosity, while the other was a rich vesicle-containing (large dense core vesicles and small clear vesicles) one. The reaction products were also found in the extracellular spaces surrounding the axons. Sometimes the reaction products were seen in the coated pits or the endocytotic vesicles of the odontoblast processes. The present study demonstrated that nerve endings (varicosities) derived from the trigeminal ganglion were present in the dentinal tubules, and that WGA-HRP extracellularly extruded from the sensory nerves in the odontoblastic layer or predentin/dentin. These findings thus suggest that sensory nerves may have some (e.g., trophic) effect on either odontoblasts or the environment around the sensory nerves in the dentin/pulp. [ABSTRACT FROM AUTHOR]

To extend our previous light microscopic observations concerning the distribution of trigeminal sensory nerves in the synovium of the rat temporomandibular joint, we investigated the detailed distribution and fine structure of sensory nerve endings at the light and electron microscopic level by the anterograde transport method using wheat germ agglutinin-horseradish peroxidase (WGA-HRP) injected into the trigeminal ganglion. At the light microscopic level, HRP-labeled nerve fibers were observed in the joint capsule and peripheral portion of the disc. The anterior portion of the disc was more densely innervated than the posterior portion, while no nerves were found in the central portion. At the electron microscopic level, HRP reaction products were observed intra-axonally in the thinly myelinated (Aδ) and unmyelinated (C) axons in the anterior portion of the joint capsule, and were also localized in the extracellular space surrounding the unmyelinated fibers and terminals. In the subsynovial layer of the synovial membrane, the majority of labeled axons located near blood vessels or among the collagenous fibrils were covered by Schwann cell sheaths, although some naked axon terminals without sheaths were also found. These unsheathed terminals contained mitochondria, small clear vesicles, and large granular vesicles, and were close to the synovial A and/or B cells near the joint cavity. The minimum distance between the terminals and synovial cells was 75 nm. This is the first demonstration of trigeminal sensory nerve terminals close to synovial lining cells or joint cavity and suggests that neuropeptides such as substance P may be released close to the synovial lining cells or joint cavity. [ABSTRACT FROM AUTHOR]

The density and distribution of substance P-like immunoreactive (SP-LI) and calcitonin gene-related peptide-like immunoreactive (CGRP-LI) nerve fibers in rat temporomandibular joint (TMJ) were investigated in whole-mount preparations and frozen sections by immunohistochemistry with the avidin-biotin-peroxidase complex method. Both types of immunoreactive nerves were observed primarily in the joint capsule, the peripheral articular disc, the synovial membrane, and the periosteum. The distribution of CGRP-LI nerves was similar to that of SPLI nerves. The anterior portion of the joint capsule and disc was most densely innervated, followed by the posterior, lateral, and medial portions. In addition, CGRP-LI nerves were more numerous and more dense in immunointensity than SP-LI nerves. In the synovial membrane, many SP- and CGRP-LI nerves terminated in the subsynovial layer, but some branches extended into the superficial synovial lining layer close to the joint cavity. Immunolabeled nerves were prominently located in the disc attachment and peripheral portion of the disc, and occasional nerves were located in the dense collagenous disc band as an actual disc. However, no fibers were detected in the central disc band. Thus, most of the disc was not innervated by any nerves. The present study provides a morphological basis for the possible roles of neuropeptides in endocytosis by synoviocytes, regulation of blood flow in the synovial membrane, nociception mechanisms of the TMJ, and modulation of the inflammatory response in the TMJ. [ABSTRACT FROM AUTHOR]

Localization of cathepsin D was studied in the junctional epithelium (JE) of healthy rat gingivae by immuno-light and -electron microscopy, by means of both the avidin-biotin-peroxidase complex method and a colloidal gold IgG method. At the light-microscopic level, cathepsin D was demonstrated in the JE and oral sulcular epithelium (OSE). Cathepsin D immunoreactivity was remarkable in the coronal portion of the JE and decreased toward its apical portion. However, cathepsin D immunoreactivity in the basal cell layer of the JE was negligible or negative. In the OSE, the granular layer was positive for cathepsin D. In the adjacent connective tissue, many macrophage-like cells (not clear at this level) close to the basal cell layer showed strong immunoreactivity. At the electron microscopic level, cathepsin D was found in the primary lysosomes and trans-cisternae of Golgi apparatus in the JE cells. These lysosomes were often fused together or were fused with cathepsin D-negative intracytoplasmic vacuoles to form secondary lysosomes, which indicated that intracellular digestion may have been in progress. However, neutrophils contained few gold particles based on cathepsin D. It is likely that the amounts of cathepsin D contained in the JE cells and macrophages are larger than those of cathepsin D contained in the neutrophils. These findings provided morphological evidence that JE cells have the same endocytotic capacity as macrophages and neutrophils, and that JE cells participate in the intracellular digestion that is carried out by lysosomal enzymes such as cathepsin D. It is suggested, in addition, that maximum intracellular digestion occurs in the coronal portion of the JE. [ABSTRACT FROM AUTHOR]

In orthodontic tooth movement (OTM), we should be concerned about external root resorption (ERR) as an undesirable iatrogenic problem, but its mechanisms are not fully understood. Since our previous epidemiologic studies found that patients with allergic diseases showed higher rates of ERR during orthodontic treatment, we explored the possible effect of allergic sensitization on ERR. In ovalbumin (OVA)-sensitized Brown-Norway rats, the amounts of ERR and OTM were greater than those in animals subjected to orthodontic force alone. The expression levels of RANKL and pro-inflammatory cytokines were increased in the periodontal tissues of sensitized rats with OTM, compared with control rats. Furthermore, leukotriene B4 (LTB4), a potent lipid mediator of allergic inflammation, and enzymes of the 5-lipoxygenase pathway, the biosynthetic pathway of leukotrienes, were also up-regulated. We found that low doses of aspirin suppressed ERR in allergen-sensitized rats, as well as the expressions of RANKL, pro-inflammatory cytokines, and LTB4. The present findings indicate that allergen sensitization has adverse effects on ERR under OTM, and that aspirin is a potential therapeutic agent for combating ERR.

Transient receptor potential (TRP) ion channels are critical contributors to the perception of various environmental stimuli. Although the oral cavity is the access point for various food types, the expression of TRP channels in oral mucosa remains unknown. We hypothesized that the oral epithelium itself may participate in sensing thermal, mechanical, and chemical conditions. The expression profiles of TRP channels exhibited regional differences among the buccal, palatal, and tongue epithelia. Changes in elevated intracellular Ca(2+) concentration ([Ca(2+)](i)) in oral epithelial cells were found after stimulation of the TRP channels with capsaicin, camphor, 4α-phorbol-12,13 didecanoate (4α-PDD), 2-aminoethoxydiphenyl borate (2-APB), and menthol. These increases in Ca(2+) appeared dependent on the TRP channels, because [Ca(2+)](i) suppression was observed after the addition of the TRPV channel antagonist ruthenium red. These results demonstrate that the oral epithelia express various TRP channels and may have functional roles in sensory activities, together with neurons.

NF-kappaB plays a pivotal role in pathogenesis in general arthritis. However, the participation of NF-kappaB in inflammation of the temporomandibular joint (TMJ) is poorly understood. We examined NF-kappaB expression in rat TMJs with synovitis induced by condyle hypermobility. By immunohistochemistry, NF-kappaB immunoreactivity was found mainly in the cytoplasm, not the nucleus, of the synovial lining cells of induced-synovitis and control TMJs. Southwestern histochemistry, a new method for detecting transcription factors, showed greater NF-kappaB expression in the nucleus of the synovial lining cells in the hypertrophic synovium than in control synovium. Increased numbers of the synovial lining cells with immunoreactivity for inducible nitric oxide synthase (iNOS), which is transcriptionally regulated by NF-kappaB, were also seen in the inflamed synovium. These findings indicate that excess mechanical stress increases NF-kappaB activation in the TMJ and suggest that active NF-kappaB is involved in the progression of TMJ inflammation.

The density and distribution of substance P-like immunoreactive (SP-LI) and calcitonin gene-related peptide-like immunoreactive (CGRP-LI) nerve fibers in rat temporomandibular joint (TMJ) were investigated in whole-mount preparations and frozen sections by immunohistochemistry with the avidin-biotin-peroxidase complex method. Both types of immunoreactive nerves were observed primarily in the joint capsule, the peripheral articular disc, the synovial membrane, and the periosteum. The distribution of CGRP-LI nerves was similar to that of SP-LI nerves. The anterior portion of the joint capsule and disc was most densely innervated, followed by the posterior, lateral, and medial portions. In addition, CGRP-LI nerves were more numerous and more dense in immuno-intensity than SP-LI nerves. In the synovial membrane, many SP- and CGRP-LI nerves terminated in the subsynovial layer, but some branches extended into the superficial synovial lining layer close to the joint cavity. Immunolabeled nerves were prominently located in the disc attachment and peripheral portion of the disc, and occasional nerves were located in the dense collagenous disc band as an actual disc. However, no fibers were detected in the central disc band. Thus, most of the disc was not innervated by any nerves. The present study provides a morphological basis for the possible roles of neuropeptides in endocytosis by synoviocytes, regulation of blood flow in the synovial membrane, nociception mechanisms of the TMJ, and modulation of the inflammatory response in the TMJ.