The Optimal Regimen for the Treatment of Temporomandibular Joint Injury Using Low-Intensity Pulsed Ultrasound in Rats with Chronic Sleep Deprivation.

Low-intensity pulsed ultrasound (LIPUS) is an emerging physical therapy for the treatment of early temporomandibular joint injury and has a good effect on promoting cartilage and subchondral bone tissue repair. However, the best LIPUS intensity and treatment duration remain unclear. This study is aimed at observing the preventive and therapeutic effects of different modes of LIPUS and at identifying the optimal LIPUS treatment regimen for temporomandibular joint injury. In the present study, rat models of temporomandibular joint injury were established using a chronic sleep deprivation (CSD) method, and the effect of LIPUS as intensities of 30, 45, and 60 mW/cm2 was observed at 7, 14, and 21 days. After CSD, the condylar cartilage of the rats demonstrated variable degrees of surface roughening, collagen fiber disarrangement or even partial exfoliation, decreased proteoglycan synthesis and cartilage thickness, decreased chondrocyte proliferation, decreased type 2 collagen (COL-2) expression, and increased matrix metalloproteinase- (MMP-) 3 expression at all three time points. When the rats with CSD received different intensities of LIPUS treatment, the pathological changes were alleviated to various extents. The groups receiving 45 mW/cm2 LIPUS showed the most significant relief of cartilage damage, and this significant effect was observed on days 14 and 21. These results demonstrated that LIPUS can effectively inhibit CSD-induced condylar cartilage damage in rats, and LIPUS treatment at an intensity of 45 mW/cm2 for at least 2 weeks is the optimal regimen for temporomandibular joint injury.

Temporomandibular joint imaging: current clinical applications, biochemical comparison with the intervertebral disc and knee meniscus, and opportunities for advancement.

Temporomandibular disorders encompass multiple pathologies of the temporomandibular joint that manifest as middle/inner ear symptoms, headache, and/or localized TMJ symptoms. There is an important although somewhat limited role of imaging in the diagnostic evaluation of temporomandibular disorders. In this manuscript, we provide a comprehensive review of TMJ anatomy, outline potentially important features of TMJ disc ultrastructure and biochemistry in comparison with the intervertebral disc and knee meniscus, and provide imaging examples of the TMJ abnormalities currently evaluable with MRI and CT. In addition, we provide an overview of emerging and investigational TMJ imaging techniques in order to encourage further imaging research based on the biomechanical alterations of the TMJ disc.

Computational characterization of the porous-fibrous behavior of the soft tissues in the temporomandibular joint.

The prevalence and severity of temporomandibular joint (TMJ) disorders have led to growing research interest in the development of new biomaterials and medical devices for TMJ implant designs. In computational designs, however, the time and stretch direction dependences of the TMJ soft tissues behavior are not considered and they are frequently based on measurements taken from non-human species or from joints that differ markedly from the human TMJ. The aim of this study was to accurately characterize the porous-fibrous properties of the TMJ soft tissues by simulating previously published experimental tests, to assist professionals in the design of new TMJ implants. To that end, material parameters were determined assuming a uniform fiber orientation throughout the entire sample. This assumption was then tested by comparing these results with those of considering multiple regions and distinct fiber orientations in each sample. Our findings validated the use of a transversely isotropic hyperelastic material model to characterize the direction dependent behavior of TMJ soft tissues and its combination with porous hyperfoam material models to mimic the compressive response of the TMJ disc. In conclusion, constitutive model proposed accurately reproduce the mechanical response of the TMJ soft tissues at different strain rates and stretch directions.

Effect of TLR4/MyD88 signaling pathway on expression of IL-1ß and TNF-α in synovial fibroblasts from temporomandibular joint exposed to lipopolysaccharide.

Accumulating evidence from previous studies suggested that interleukin-1 (IL-1ß) and tumor necrosis factor-α (TNF-α) play an important role in pathogenesis of temporomandibular disorders (TMD). However, the cell surface receptors and the intracellular signal pathways leading to these cytokines expression are not fully understood. In the current study, we investigated the roles of Toll-like receptor 4 (TLR4) and its adaptor myeloid differentiation factor 88 (MyD88) in the expression of IL-1ß and TNF-α in synovial fibroblasts (SFs) separated from rat temporomandibular joint (TMJ) with lipopolysaccharide (LPS) stimulation. The results showed that treatment with LPS could increase TLR4, MyD88, IL-1ß, and TNF-α expression at both mRNA and protein levels. In addition, increased expression of IL-1ß and TNF-α could be blocked by treatment with TAK-242, a blocker of TLR4 signaling, and also by MyD88 inhibitory peptide (MIP). These findings suggested that maybe TLR4/MyD88 signal transduction pathway participates in enhanced expression of IL-1 and TNF-α in patients with TMD. The activation of TLR4/MyD88 signal transduction pathway which results in production of proinflammatory factors may play a role in the pathogenesis of TMD.

Cytokines in healthy temporomandibular joint synovial fluid.

Analysis of temporomandibular joint (TMJ) synovial fluid may elucidate the aetiology of temporomandibular disorders and arthritic conditions, as well as the inflammatory mechanisms involved. Knowledge about healthy synovial fluid is necessary to understand TMJ pathologies. We aimed to quantify the proinflammatory cytokines interleukin (IL)-1ß, IL-2, IL-6 and tumour necrosis factor (TNF), and the anti-inflammatory cytokines IL-10 and interferon (IFN)-γ in healthy TMJ synovial fluid to serve as reference values for future studies on TMJ pathologies. Twenty healthy, young adult volunteers without temporomandibular dysfunction were included. Bilateral synovial fluid samples were obtained using the push-pull technique with hydroxocobalamin described by Alstergren in 1999. Cytokines were quantified with Luminex multiplex assays and compared using nonparametric statistical analysis. No serious adverse effects were reported. Of 40 possible samples, 14 fulfilled the strict sampling criteria and were included in the analysis. Cytokine values (reported as medians with interquartile ranges) were as follows: TNF, 23 (13-37) pg mL(-1) ; IL-2, 1·8 (0-22) pg mL(-1) ; and INF-γ, 10 (0-47) pg mL(-1) . IL-1ß, IL-6 and IL-10 were almost undetectable. In addition, TNF and INF-γ cytokine levels correlated. We demonstrated that TNF was consistently detected and IFN-γ and IL-2 sporadically detected in the TMJ synovial fluid of healthy individuals using the hydroxocobalamin method and a multiplex assay. The cytokines IL-10, IL-1ß and IL-6 were barely detectable in this sample of healthy TMJs.

Características fisicoquímicas e citológicas do líquido sinovial da articulação temporomandibular em eqüinos / Physical, biochemical and cytological characteristics of the equine temporomandibular joint synovial fluid

Foram estudadas as características fisicoquímicas e citológicas do líquido sinovial da articulação temporomandibular de dez eqüinos hígidos. Verificou-se que o líquido é viscoso, amarelo claro a citrino, límpido e livre de partículas à temperatura ambiente. Houve contaminação da amostra por sangue em três amostras que se apresentaram amarelo avermelhadas a vermelhas e de aspecto turvo. A taxa de glicose variou entre 100 e 250 e a concentração protéica não ultrapassou 3,8g/dL. O número médio de células nucleadas foi de 417 células/µL, com predominância de grandes células mononucleares e linfócitos. As mensurações das características pesquisadas no líquido sinovial da articulação temporomandibular de eqüinos são de execução simples e passíveis de implantação na rotina de atendimentos clínico-cirúrgicos.

Physical, biochemical and cytological characteristics of the temporomandibular joint synovial fluid were studied in ten clinically normal horses. It is a viscous, pale yellow, clear fluid and without flocculent material at room temperature. There was blood contamination in three samples, they presented red-yellow to red and cloudy. The range of glucose levels were 100 to 250 and its protein concentration was up to 3,8g/dL. Nucleated cells mean number was 417 cells/µL, with predominating large mononuclear cells and lymphocytes. Equine temporomandibular synovial fluids can be easily evaluated, being feasible in clinical and surgical routine, and the information may be useful to the diagnosis, treatment and prognosis of animals with temporomandibular alterations.

Tumoral calcium pyrophosphate dihydrate crystal deposition disease of the temporomandibular joint: identification on crystallography.

This paper reports a case of calcium pyrophosphate dihydrate (CPPD) crystal deposition in the temporomandibular joint (TMJ) of a 59-year-old man with the chief complaint of severe pain in the left TMJ. On CT a radiopaque area was seen around the condylar process of the left TMJ with irregular destructive bony changes. A provisional diagnosis of crystalline-induced arthritis was made on histopathology of a biopsy specimen. Electron probe microanalysis (EPMA), scanning electron microscopy (SEM) and X-ray diffraction showed both CPPD and hydroxyapatite (HA) in the crystalline materials. Identification of these two types of crystal in crystal deposition disease of TMJ, using crystallography, is discussed.

Histological investigation of osseous changes of mandibular condyles with backscattered electron images.

OBJECTIVES: This study aims to confirm the radiographic morphology of mandibular condyles with microscopic observation and to investigate the mechanism of osseous changes of adult human mandibular condyles. METHODS: Following the radiographic examinations of ten temporomandibular joints (TMJs) from five cadavers using conventional tomography, helical CT and limited cone beam CT for dentistry (limited CBCT), ten mandibular condyles were removed. Micro CT images were made of the undecalcified dissected joints in the sagittal plane, perpendicular to the long axis of the condyles, to produce images similar in orientation to the radiographic images. Tissue morphology was observed through backscattered electron images and elemental analysis using electron probe microanalysis (EPMA). The microscopic findings were compared with the radiographic findings. RESULTS: Limited CBCT images most accurately depicted erosive change of the bone cortex of the mandibular condyle. EPMA indicated bone remodelling in all mandibular condyles analysed, which showed cartilage calcification different from typical enchondral ossification. No finding of dissolution or disruption of the superficial soft tissue was observed in all specimens, including those presenting radiographic findings of osteoarthrosis. Active bone remodelling with hypertrophic changes of superficial soft tissue was observed in mandibular condyles, which implied radiographic findings of osteoarthrosis. CONCLUSIONS: The high detectability of limited CBCT images on bony morphology of mandibular condyles was confirmed. It is speculated that bone remodelling of adult human mandibular condyles is brought about by cartilage calcification. It was suggested that the osseous changes classified as osteoarthrosis with radiographic images are normal bone remodelling.

Analysis of estrogen binding sites of the posterior ligament of the human TMJ.

BACKGROUND: Conflicting results have been reported regarding the presence or absence of estrogen-binding sites (EBS) in the human temporomandibular joint (TMJ). The possible role of female sex hormones in the pathophysiology of internal derangement of the TMJ has been suggested to explain the prevalence of TMJ symptoms in female patients. PATIENTS AND METHODS: Posterior bilaminar tissue excised during TMJ articular disc repositioning and posterior ligament repair was taken from 28 patients (26 female, 2 male) for evaluation. Cryosections were stained using a monoclonal antibody (Mab) against EBS. To ensure efficacy of the antibody staining procedure, an internal positive control consisting of human breast tissue previously proven EBS-positive was used. No asymptomatic control TMJ tissue was available for our study. RESULTS: None (0%) of 28 TMJ tissue specimens showed nuclear-staining positive for the presence of EBS in the posterior bilaminar tissue of the TMJ. However, estrogen-binding sites associated with probable inflammatory cells were observed. Our results are consistent with the probability of positives as high as 0.1234 using a 95% confidence interval. CONCLUSIONS: The lack of EBS of the posterior ligament of the TMJ suggests that the role of estrogen contributing to internal derangement of the TMJ appears not to be significant.

Expression of caveolin-3 in the fibroblast-like type B synoviocytes in the rat temporomandibular joint.

The present study revealed that the fibroblast-like type B synoviocytes (covering the surface of the synovial membrane in the rat temporomandibular joint) had muscle-specific caveolin-3 protein in their caveolae. The existence of two kinds of type B synoviocytes (with and without caveolin-3-immunoreactions even in the synovial lining layer) might reflect the functional difference between them.

Localization of CD44 and hyaluronan in the synovial membrane of the rat temporomandibular joint.

Previous studies have pointed out a lack of adhesion structures in the synovial lining layer of the rat temporomandibular joint (TMJ) despite showing an epithelial arrangement. CD44, a major cell adhesion molecule, plays crucial roles as an anchor between cells and extracellular matrices by binding hyaluronan (HA) for the development of organs or the metastasis of tumors. The present study examined the localization of CD44 in the synovial membrane of the rat TMJ by immunocytochemistry for OX50, ED1, and Hsp25, which are markers for the rat CD44, macrophage-like type A, and fibroblast-like type B synoviocytes, respectively. Histochemistry for HA-binding protein (HABP) was also employed for the detection of HA. OX50 immunoreactions were found along the cell surface and, in particular, accumulated along the surface of the articular cavity. Observations by a double immunostaining and immunoelectron microscopy revealed that all the OX50-immunopositive cells were categorized as fibroblastic type B cells, which had many caveolae and a few vesicles reactive to intense OX50. However, the macrophage-like type A cells did not have any OX50 immunoreaction in the synovial lining layer. A strong HABP reaction was discernable in the extracellular matrix surrounding both OX50-positive and -negative cells in the synovial lining layers, exhibiting a meshwork distribution, but weak in its sublining layer. This localization pattern of CD44 and HABP might be involved in the formation of the epithelial arrangement of the synovial lining layer. Furthermore, OX50 immunonegativity in the type A cells suggests their low phagocytotic activity in the rat TMJ under normal conditions.

Immunocytochemical localization of MAPKAPK-2 and Hsp25 in the rat temporomandibular joint.

One series of our research has shown an intense expression of immunoreaction for heat shock protein 25 (Hsp25) in various cellular elements in the rat temporomandibular joint (TMJ). This protein is the major substrate of mitogen-activated protein kinase-activated protein kinase-2 (MAPKAPK-2), which mediates an intracellular stress-activated signaling pathway to stimulate cytosolic actin reorganization under various stresses. The present study was undertaken to examine the localization of MAPKAPK-2 in the rat TMJ by immunocytochemical techniques. Furthermore, confocal microscopy with double staining was employed to demonstrate the colocalization of MAPKAPK-2 and Hsp25. Immunocytochemistry for MAPKAPK-2 showed an intense immunoreaction in the cytoplasm of the synovial lining cells, the endothelial cells, and the fibroblasts in the synovial membrane of the rat TMJ. Double immunostaining under a confocal microscope succeeded in demonstrating the colocalization of MAPKAPK-2 and Hsp25 immunoreactions in the cytoplasm of fibroblastic type B synoviocytes in the TMJ. On the other hand, the macrophage-like type A-cells expressed MAPKAPK-2 immunoreactions but lacked Hsp25 immunoreactivity. The cells in the articular disk and the chondrocytes in the maturative and hypertrophic layer of the mandibular cartilage also showed intense immunoreactions for MAPKAPK-2 and Hsp25. In addition to cytoplasmic localization, MAPKAPK-2 immunoreactions were found in the nucleus of some synovial lining cells, cells in the articular disk, and chondrocytes. Current observations imply the presence of the phosphorylation of Hsp25 via activated MAPKAPK-2 in the cytoplasm. MAPKAPK-2 and Hsp25 possibly participate in the induction of cytoskeletal changes to the various cellular elements in rat TMJ under normal conditions.

The immunohistochemical distribution of vimentin in human temporomandibular joint samples.

The aim of this investigation was to evaluate the immunohistochemical distribution of vimentin in the temporomandibular joint (TMJ) and to compare it with the control specimens. Immunohistochemical distribution in the disc and synovial membrane in 30 human TMJ (internal derangement of TMJ, n = 20; and control, n = 10) was studied immunohistologically using paraffin-embedded tissue and specific anti-human vimentin monoclonal antibody. Vimentin expression was distributed in chondrocyte-like cells, synovial cells and endothelial cells. There was an obvious distinction of vimentin immunoreactivity between the control specimens and internal derangement cases, in the posterior and/or anterior loose connective tissues. In particular, intensive vimentin expression was detected in the hypertrophic synovial membrane of internal derangement cases. The findings of the present study suggest that vimentin might be an important marker of pathological hypertrophy of the synovial membrane and/or connective tissue with internal derangement of TMJ.

Soluble tumour necrosis factor receptors in synovial fluids from temporomandibular joints with painful anterior disc displacement without reduction and osteoarthritis.

The objective of this study was to detect soluble-form tumour necrosis factor receptors (sTNFRs) in temporomandibular joint (TMJ) synovial fluid aspirates, and to compare the sTNFR concentrations between painful anterior disc displacement without reduction and osteoarthritis (ADDwoR/OA) and asymptomatic TMJs. Synovial fluid was sampled from the superior TMJ cavity of 11 painful ADDwoR/OA cases (mean age: 36.9 years) and 10 asymptomatic females (mean age: 24.7 years) by diluted aspiration. The concentrations of sTNFR-I and -II in the synovial fluid were measured using human sTNFR-I and -II enzyme-linked immunosorbent assays. The total protein concentrations in synovial fluids were measured using a bicinchoninic acid protein assay kit. All data were normalised to the total protein concentration of each sample.Two-way factorial analysis of variance and post hoc multiple comparison revealed that: (1). mean normalised sTNFR-I and -II concentrations were higher in TMJ synovial aspirates from ADDwoR/OA patients than from healthy controls; (2). in the ADDwoR/OA patients and the healthy controls, the sTNFR-I concentration in TMJ synovial aspirates was higher than the sTNFR-II concentration; and (3). high TMJ synovial aspirate sTNFR-II seemed to be associated with less TMJ pain and a less restricted range of mouth opening in the ADDwoR/OA patients. The concentrations of sTNFRs in TMJ synovial fluid are higher in the presence of painful ADDwoR/OA, which could modulate intracapsular inflammation.

[Effects of unilateral mandibular distraction osteogenesis on temporomandibular joint].

OBJECTIVE: To study the effects of unilateral mandibular distraction osteogenesis(MDO) on the temporomandibular joint(TMJ). METHODS: Ten rhesus monkeys were used to perform right MDO. After 5 days, latency period, the distractor was initiated at the rate of 0.5 mmx2/day, totally 15 days. When MDO completed, and 2,4,6,and 12 weeks after MDO, X-ray films of bilateral TMJ were taken and one group of animals was sacrificed. Bilateral TMJ were obtained and paraffin slices were made HE and immunohistochemistry stains of TGF-beta1 were done on the slices. RESULTS: After MDO, the monkeys' mandible midline obviously deflected to the left side. The front teeth had open bite and reverse bite. The right molars were severely open bite and the left molars were reverse bite. From the X-ray film, we can see that the right condylar process moved backward and upward in the acetabulum, while the left condylar process moved forward and downward. On the slices, both sides of the TMJ showed different degrees of degeneration and proliferation, but the changes on the right side were more severe and obvious than the left side, and had involved all the cartilage's layers and the bone tissues under the cartilage. In some animals, there even appeared cartilage erosion and light histological changes of acetabulum. During the consolidation period, the open bite gradually closed, and the condylar process gradually moved to normal position. Twelve weeks after MDO, the open bite at the right side disappeared, but the front teeth open bite and reverse bite still existed. The right condylar process still lay a little backward and had some degree of degeneration. The left TMJ had the normal position and histology except for cartilage hypertrophy layer thickness. At different times during consolidation period, the positive stains of TGF-beta1 in condylar process on both sides were all more obvious than that of the control group. CONCLUSION: MDO can cause TMJ position change and light histological degeneration, but these changes can gradually rehabilitate in the future. TGF-beta1 may play a role in the reconstructive process of TMJ degeneration.

mu-Opioid receptor mRNA expression and immunohistochemical localization in the rat temporomandibular joint.

This study was undertaken to examine the presence and distribution of the mu-opioid receptor (MOR) in the non-inflamed rat temporomandibular joint (TMJ) using non-radiographic in situ hybridization at the mRNA level and immunohistochemistry at the protein level. MOR mRNA and MOR-like immunoreactivity (MOR-LI) were found around the small blood vessels in the anterior part of the synovial membrane. The number of MOR mRNA signals in the anterior synovial membrane was significantly higher than that in the posterior part. Morphologically, MOR mRNA and MOR-LI were localized in amorphous materials considered to be nervous tissue, as well as some cell types considered to be macrophages, mast cells and endothelial cells. The present study showed the distribution of MOR in the rat TMJ synovial membrane and suggests that the opiate system plays an important role in endogenous analgesia in the TMJ.

Arrangement of D-periodic collagen fibrils and association of proteoglycans with fibrils in the synovium of the mouse temporomandibular joint.

The present study was performed to examine changes in the arrangement of D-periodic collagen fibrils in the synovium of the growing temporomandibular joint in mice. At 1 week of age, the mandibular condyle was undeveloped, and only a few collagen fibrils were recognizable in the subintimal layer of the synovium. At 8 weeks, the mandibular condyle was structurally developed with an increase of collagen fibrils in the synovium; a fully mature condyle was observed at 6 months of age. The close association of proteoglycans with collagen fibrils in the synovium of the growing joint was examined by both conventional and energy-filtering transmission electron microscopy of cupromeronic blue-stained specimens. Proteoglycans were associated with D-periodic collagen fibrils in the short filamentous form in random fashion at 1 week of age, but in a regular pattern with D-periodicity at 6 months. These associations in the synovium could be correlated with the mechanical character of the temporomandibular joint.

Gender-specific differences in temporomandibular retrodiscal tissues of the goat.

Healthy, adult, male and female goat temporomandibular retrodiscal tissues were characterized to determine if biochemical differences existed between the genders. RNA concentrations were not different between male and female retrodiscal tissues; however, the DNA concentration in female retrodiscal tissues was 82% greater than in male retrodiscal tissues. Collagen concentrations were significantly greater in male retrodiscal tissues, and this was reflected in significant gender differences of type I and III collagen concentrations. More specifically, male temporomandibular retrodiscal tissues contained 70% more type I collagen and 119% more type III collagen when compared to female retrodiscal tissues. These differences in collagens and DNA reflect a gender difference in temporomandibular retrodiscal tissue composition that underlies divergent biomechanical and neurophysiological properties.

Age-associated changes in decorin in rat mandibular condylar cartilage.

The small proteoglycan decorin strongly binds the fibrils of collagen types I and II; this interaction is thought to play a part in the maintenance of tissue integrity and biomechanical properties. In limb articular cartilage, there is evidence that decorin synthesis increases with age and that it is elevated in response to increased loading or in osteoarthritic cartilage. The aim here was to characterize the presence and relative amount of decorin in the condylar cartilage of the temporomandibular joint (TMJ) with maturation by Western blotting, and to assess its tissue localization by immunohistochemistry. Comparative data were obtained from tibial articular cartilage, which has been extensively studied. Cartilage from the mandibular condyle and tibial plateau was harvested from 24-day-old (growing) and 161-day-old (young adult) female Sprague-Dawley rats. In growing animals, decorin appeared slightly more abundant in the mandibular condylar cartilage than in articular cartilage, whereas in young adult animals the decorin content in the TMJ cartilage was noticeably less than in limb articular cartilage. Although there was an increase in decorin abundance with age at the TMJ, the increase in decorin with age in limb articular cartilage was considerably more pronounced. These data indicate that, although decorin is present in mandibular condylar cartilage, its abundance in adults is less than in limb articular cartilage; thus, maturation-associated changes may be dissimilar in magnitude from those documented for limb articular cartilage.