The Roles of Indian Hedgehog Signaling in TMJ Formation.

The temporomandibular joint (TMJ) is an intricate structure composed of the mandibular condyle, articular disc, and glenoid fossa in the temporal bone. Apical condylar cartilage is classified as a secondary cartilage, is fibrocartilaginous in nature, and is structurally distinct from growth plate and articular cartilage in long bones. Condylar cartilage is organized in distinct cellular layers that include a superficial layer that produces lubricants, a polymorphic/progenitor layer that contains stem/progenitor cells, and underlying layers of flattened and hypertrophic chondrocytes. Uniquely, progenitor cells reside near the articular surface, proliferate, undergo chondrogenesis, and mature into hypertrophic chondrocytes. During the past decades, there has been a growing interest in the molecular mechanisms by which the TMJ develops and acquires its unique structural and functional features. Indian hedgehog (Ihh), which regulates skeletal development including synovial joint formation, also plays pivotal roles in TMJ development and postnatal maintenance. This review provides a description of the many important recent advances in Hedgehog (Hh) signaling in TMJ biology. These include studies that used conventional approaches and those that analyzed the phenotype of tissue-specific mouse mutants lacking Ihh or associated molecules. The recent advances in understanding the molecular mechanism regulating TMJ development are impressive and these findings will have major implications for future translational medicine tools to repair and regenerate TMJ congenital anomalies and acquired diseases, such as degenerative damage in TMJ osteoarthritic conditions.

Roles of Ihh signaling in chondroprogenitor function in postnatal condylar cartilage.

Condylar articular cartilage in mouse temporomandibular joint develops from progenitor cells near the articulating surface that proliferate, undergo chondrogenesis and mature into hypertrophic chondrocytes. However, it remains unclear how these processes are regulated, particularly postnatally. Here we focused on the apical polymorphic layer rich in progenitors and asked whether the phenotype and fate of the cells require signaling by Indian hedgehog (Ihh) previously studied in developing long bones. In condyles in newborn mice, the apical polymorphic/progenitor cell layer was ~10 cell layer-thick and expressed the articular matrix marker Tenascin-C (Tn-C), and the underlying thick cell layer expressed Tn-C as well as the chondrogenic master regulator Sox9. By 1 month, condylar cartilage had gained its full width, but became thinner along its main longitudinal axis and displayed hypertrophic chondrocytes. By 3 months, articular cartilage consisted of a 2-3 cell layer-thick zone of superficial cells and chondroprogenitors expressing both Tn-C and Sox9 and a bottom zone of chondrocytes displaying vertical matrix septa. EdU cell tracing in juvenile mice revealed that conversion of chondroprogenitors into chondrocytes and hypertrophic chondrocytes required about 48 and 72 h, respectively. Notably, EdU injection in 3 month-old mice labeled both progenitors and maturing chondrocytes by 96 h. Conditional ablation of Ihh in juvenile/early adult mice compromised chondroprogenitor organization and function and led to reduced chondroprogenitor and chondrocyte proliferation. The phenotype of mutant condyles worsened over time as indicated by apoptotic chondrocyte incidence, ectopic chondrocyte hypertrophy, chondrocyte column derangement and subchondral bone deterioration. In micromass cultures of condylar apical cells, hedgehog (Hh) treatment stimulated chondrogenesis and alkaline phosphatase (APase) activity, while treatment with HhAntag inhibited both. Our findings indicate that the chondroprogenitor layer is continuously engaged in condylar growth postnatally and its organization and functioning depend on hedgehog signaling.

Biomechanical properties of murine TMJ articular disc and condyle cartilage via AFM-nanoindentation.

This study aims to quantify the biomechanical properties of murine temporomandibular joint (TMJ) articular disc and condyle cartilage using AFM-nanoindentation. For skeletally mature, 3-month old mice, the surface of condyle cartilage was found to be significantly stiffer (306±84kPa, mean±95% CI) than those of the superior (85±23kPa) and inferior (45±12kPa) sides of the articular disc. On the disc surface, significant heterogeneity was also detected across multiple anatomical sites, with the posterior end being the stiffest and central region being the softest. Using SEM, this study also found that the surfaces of disc are composed of anteroposteriorly oriented collagen fibers, which are sporadically covered by thinner random fibrils. Such fibrous nature results in both an F-D3/2 indentation response, which is a typical Hertzian response for soft continuum tissue under a spherical tip, and a linear F-D response, which is typical for fibrous tissues, further signifying the high degree of tissue heterogeneity. In comparison, the surface of condyle cartilage is dominated by thinner, randomly oriented collagen fibrils, leading to Hertzian-dominated indentation responses. As the first biomechanical study of murine TMJ, this work will provide a basis for future investigations of TMJ tissue development and osteoarthritis in various murine TMJ models.

Limitations of the TheraMon® -microsensor in monitoring occlusion therapy.

PURPOSE: A proof of principle study showed that the TheraMon® -microsensor can reliably measure wearing times of glasses and occlusion patches while having no discomfort for the bearer. The goal of this study was to identify specific temperature profiles in a larger patient group wearing the microsensor with an occlusion patch on the eye, the forehead or in the trouser pocket. METHODS: Temperature measurements with microsensors were performed at the orbital rim of the eye, at the forehead and in a trousers pocket of 20 healthy adults. The temperature measurements were analysed comparing the three locations using a repeated-measures anova with a 0.050 two-sided significance level. RESULTS: There was no significant temperature difference in wearing the microsensors at the orbital rim compared to forehead (p = 0.507). A significant difference could be detected if the microsensors had been in trousers pockets as compared to being positioned at orbital rim/forehead (p = 0.001). The study participants reported easy handling and comfortable wearing. CONCLUSION: This study demonstrates for the first time that TheraMon® -microsensors are fully operational in monitoring occlusion therapy in a larger group of patients (20 adult study participants). Because of the microsensor's ease of handling further studies in a larger group of children is recommended.

Osteophyte formation and matrix mineralization in a TMJ osteoarthritis mouse model are associated with ectopic hedgehog signaling.

The temporomandibular joint (TMJ) is a diarthrodial joint that relies on lubricants for frictionless movement and long-term function. It remains unclear what temporal and causal relationships may exist between compromised lubrication and onset and progression of TMJ disease. Here we report that Proteoglycan 4 (Prg4)-null TMJs exhibit irreversible osteoarthritis-like changes over time and are linked to formation of ectopic mineralized tissues and osteophytes in articular disc, mandibular condyle and glenoid fossa. In the presumptive layer of mutant glenoid fossa's articulating surface, numerous chondrogenic cells and/or chondrocytes emerged ectopically within the type I collagen-expressing cell population, underwent endochondral bone formation accompanied by enhanced Ihh expression, became entrapped into temporal bone mineralized matrix, and thereby elicited excessive chondroid bone formation. As the osteophytes grew, the roof of the glenoid fossa/eminence became significantly thicker and flatter, resulting in loss of its characteristic concave shape for accommodation of condyle and disc. Concurrently, the condyles became flatter and larger and exhibited ectopic bone along their neck, likely supporting the enlarged condylar heads. Articular discs lost their concave configuration, and ectopic cartilage developed and articulated with osteophytes. In glenoid fossa cells in culture, hedgehog signaling stimulated chondrocyte maturation and mineralization including alkaline phosphatase, while treatment with hedgehog inhibitor HhAntag prevented such maturation process. In sum, our data indicate that Prg4 is needed for TMJ integrity and long-term postnatal function. In its absence, progenitor cells near presumptive articular layer and disc undergo ectopic chondrogenesis and generate ectopic cartilage, possibly driven by aberrant activation of Hh signaling. The data suggest also that the Prg4-null mice represent a useful model to study TMJ osteoarthritis-like degeneration and clarify its pathogenesis.

Effect of cephalograms on decisions for early orthodontic treatment.

OBJECTIVE: To assess the extent to which cephalograms modify therapeutic decisions related to early orthodontic treatment. MATERIALS AND METHODS: Diagnostic records of six patients requiring early treatment were digitized for this purpose. A total of 234 orthodontists were then asked to select therapeutic measures on a treatment-planning sheet at two different times (T1 and T2). Three groups of orthodontists were formed and were provided with case-specific records either including or not including cephalograms and the appended tracing values. Forty-seven orthodontists completed all phases of the questions. RESULTS: Statistical analysis revealed only two statistically significant differences between the first (T1) and second (T2) treatment plans, both concerning options of transverse treatment. None of the other results showed any significant changes from T1 to T2 treatment planning. CONCLUSION: It follows that cephalograms did not influence therapeutic decisions for early orthodontic treatment and should not be routinely used in very young patients.

Measuring wearing times of glasses and ocular patches using a thermosensor device from orthodontics.

PURPOSE: Amblyopia is one of the most common visual disorders in children. The risk of severe visual impairment on the healthy eye is doubled in patients with amblyopia. If detected early enough, the chances of visual rehabilitation are good. Treatment consists of refractive correction and occlusion of the dominant eye. Patient compliance is an important factor and can be monitored using thermosensors. It was the goal of our study to give proof of the principle that the wearing times of glasses and patches can be measured using a comparatively small and commercially available microsensor. METHODS: Agreement between wearing times protocols of ocular patching/refractive correction and temperature measurements of thermosensors attached to the patches or glasses of three individuals were analysed using the Bland-Altman method. It was also analysed whether blinded persons could distinguish between temperature curves of patches and glasses, or temperature curves of an incubator or while worn in a pocket. RESULTS: The temperatures picked up by the microsensors indicate the beginning and the end wearing times of either glasses or ocular patches through steep temperature difference and a distinct temperature curve during measurements. Although blinded test persons were able to cleary distinguish between temperature profiles from incubator/pocket measurements compared to glasses/patching, glasses and patching curves could be discriminated correctly in only 50%. Differences between wearing time protocols and temperature measurements were within the limits of agreement as stated by the Bland-Altman plots. CONCLUSION: The TheraMon(®) microsensor can reliably measure wearing times of glasses and ocular patches without making the wearer uncomfortable, although the data are not unquestionable, especially in higher surrounding temperatures. Further studies on a larger number of individuals with different wearing profiles are needed.

Distalization pattern of the maxillary arch depending on the number of orthodontic miniscrews.

OBJECTIVE: To determine the effects of linear force vector(s) from interradicular miniscrews on the distalization pattern of the maxillary arch in adult Class II patients. MATERIALS AND METHODS: Twenty-five adult patients with mild to moderate Class II dentition and minimal crowding were collected. Either single (group A, n  =  12) or dual (group B, n  =  13) miniscrews were inserted on the posterior interradicular area to deliver a distalizing force to the main archwire. The displacement patterns of maxillary incisors and molars were measured and compared. RESULTS: Significant distalization in the molars and incisors was shown in both groups. Significantly greater distalization and intrusion of the first molar and intrusive displacement of the incisor, together with significant reduction of the mandibular plane, were noted in group B, in contrast to the rotation of the occlusal plane in group A. CONCLUSIONS: Interradicular miniscrews predictably induced total arch distalization, leading to the correction of Class II. Additional miniscrews in the premolar area appear to facilitate intrusion and distalization of the entire arch according to the position of the force vectors.