Nobuhiko Kawai

Lubrication of the temporomandibular joint.

Although tissue engineering of the temporomandibular joint (TMJ) structures is in its infancy, tissue engineering provides the revolutionary possibility for treatment of temporomandibular disorders (TMDs). Recently, several reviews have provided a summary of knowledge of TMJ structure and function at the biochemical, cellular, or mechanical level for tissue engineering of mandibular cartilage, bone and the TMJ disc. As the TMJ enables large relative movements, joint lubrication can be considered of great importance for an understanding of the dynamics of the TMJ. The tribological characteristics of the TMJ are essential for reconstruction and tissue engineering of the joint. The purpose of this review is to provide a summary of advances relevant to the tribological characteristics of the TMJ and to serve as a reference for future research in this field. This review consists of four parts. Part 1 is a brief review of the anatomy and function of the TMJ articular components. In Part 2, the biomechanical and biochemical factors associated with joint lubrication are described: the articular surface topology with microscopic surface roughness and the biomechanical loading during jaw movements. Part 3 includes lubrication theories and possible mechanisms for breakdown of joint lubrication. Finally, in Part 4, the requirement and possibility of tissue engineering for treatment of TMDs with degenerative changes as a future treatment regimen will be discussed in a tribological context.

The frictional coefficient of the temporomandibular joint and its dependency on the magnitude and duration of joint loading.

In synovial joints, friction between articular surfaces leads to shear stress within the cartilaginous tissue, which might result in tissue rupture and failure. Joint friction depends on synovial lubrication of the articular surfaces, which can be altered due to compressive loading. Therefore, we hypothesized that the frictional coefficient of the temporomandibular joint (TMJ) is affected by the magnitude and duration of loading. We tested this by measuring the frictional coefficient in 20 intact porcine TMJs using a pendulum-type friction tester. The mean frictional coefficient was 0.0145 (SD 0.0027) after a constant loading of 50 N during 5 sec. The frictional coefficient increased with the length of the preceding loading duration and exceeded 0.0220 (SD 0.0014) after 1 hr. Application of larger loading (80 N) resulted in significantly larger frictional coefficients. In conclusion, the frictional coefficient in the TMJ was proportional to the magnitude and duration of joint loading.

Dynamic Shear Properties of the Temporomandibular Joint Disc

Shear stress might be an important factor associated with fatigue failure and damage of the temporomandibular joint disc. Little information, however, is available on the dynamic behavior of the disc in shear. Since the disc is an anisotropic and viscoelastic structure, in the present study the dependency of the dynamic shear behavior on the direction and frequency of loading was examined. Ten porcine discs were used for dynamic shear tests. Shear stress was applied in both anteroposterior (A-P test) and mediolateral (M-L test) directions. The dynamic moduli increased as the loading frequency increased. The dynamic elasticity was significantly larger in the A-P test than in the M-L test, although the dynamic viscosity was similar in both tests. The present results suggest that non-linearities, compression/shear coupling, and intrinsic viscoelasticity affect the shear material behavior of the disc, which might have important implications for the transmission of load in the temporomandibular joint.

Fibre-type composition of rabbit jaw muscles is related to their daily activity.

Skeletal muscles contain a mixture of fibres with different contractile properties, such as maximum force, contraction velocity and fatigability. Muscles adapt to altered functional demands, for example, by changing their fibre‐type composition. This fibre‐type composition can be changed by the frequency, duration and presumably the intensity of activation. The aim of this study was to analyse the relationship between the spontaneous daily muscle activation and fibre‐type composition in rabbit jaw muscles. Using radio‐telemetry combined with electromyography, the daily activity of five jaw muscles was characterized in terms of the total duration of muscle activity (duty time) and the number of activity bursts. Fibre‐type composition of the muscles was classified by analysing the myosin heavy chain content of the fibres. The amount of slow‐type fibres was positively correlated to the duty time and the number of bursts only for activations exceeding 20–30% of the maximum activity per day. Furthermore, cross‐sectional areas of the slow‐type fibres were positively correlated to the duty time for activations exceeding 30% of the maximum activity. The present data indicate that the amount of activation above a threshold (> 30% peak activity) is important for determining the fibre‐type composition and cross‐sectional area of slow‐type fibres of a muscle. Activation above this threshold occurred only around 2% of the time in the jaw muscles, suggesting that contractile properties of muscle fibres are maintained by a relatively small number of powerful contractions per day.

Heterogeneity of fiber characteristics in the rat masseter and digastric muscles

The functional requirements in muscle use are related to the fiber type composition of the muscles and the cross‐sectional area of the individual fibers. We investigated the heterogeneity in the fiber type composition and fiber cross‐sectional area in two muscles with an opposing function, namely the digastric and masseter muscles (n = 5 for each muscle) of adult male rats, by means of immunohistochemical staining according to their myosin heavy chain (MyHC) content. The digastric and masseter muscles were taken from Wistar strain male rats 10 weeks old. In the masseter six predefined sample locations were examined; in the digastric four. Most regions showed dominant proportions of type IIA and IIX fibers. However, both muscles also revealed a regional heterogeneity in their fiber type distribution. In the digastric, type I fibers were detected only at the central and deep areas of the anterior and posterior belly, respectively. Meanwhile, the peripheral area of the anterior belly contained a higher proportion of type IIB fibers. In the masseter, the type I fibers were absent. In the superficial masseter the distribution of IIA and IIB fibers was significantly different between the superior and inferior regions. In the deep masseter, regional differences were observed among all four examined areas, of which the posterolateral region contained the highest proportion of type IIB fibers. The cross‐sectional areas of type IIB fibers were always the largest, followed by the type IIX and IIA fibers. Only a few differences in cross‐sectional area of corresponding fiber types were detected between the various sites. In conclusion, the masseter and digastric muscles showed an obvious heterogeneity of fiber type composition and fiber cross‐sectional area. Their heterogeneity reflects the complex role of the both muscles during function. This detailed description of the fiber type composition can serve as a reference for future studies examining the muscular adaptations after the onset of various diseases in the masticatory system.

Shear Properties of the Temporomandibular Joint Disc in Relation to Compressive and Shear Strain

Shear stress can result in fatigue, damage, and irreversible deformation of the temporomandibular joint disc. Insight into the dynamic shear properties of the disc may give insight into the mechanism inducing tissue failure due to shear. We tested the hypothesis that the dynamic shear properties of the disc depend on the amount of shear and compressive strain. Twenty-four porcine discs were used for dynamic shear tests. The specimens were clamped between the plates of a loading apparatus under compressive strains of 5%, 10%, and 15%. Dynamic shear was applied to the specimen by a sinusoidal strain of, respectively, 0.5%, 1.0%, and 1.5%. Both the dynamic elasticity and viscosity were proportional to compressive strain and inversely proportional to shear strain. These shear characteristics suggest a significant role of compressive and shear strain on the internal friction of the disc.

Adaptation of rat jaw muscle fibers in postnatal development with a different food consistency: an immunohistochemical and electromyographic study

The development of the craniofacial system occurs, among other reasons, as a response to functional needs. In particular, the deficiency of the proper masticatory stimulus affects the growth. The purpose of this study was to relate alterations of muscle activity during postnatal development to adaptational changes in the muscle fibers. Fourteen 21‐day‐old Wistar strain male rats were randomly divided into two groups and fed on either a solid (hard‐diet group) or a powder (soft‐diet group) diet for 63 days. A radio‐telemetric device was implanted to record muscle activity continuously from the superficial masseter, anterior belly of digastric and anterior temporalis muscles. The degree of daily muscle use was quantified by the total duration of muscle activity per day (duty time), the total burst number and their average length exceeding specified levels of the peak activity (5, 20 and 50%). The fiber type composition of the muscles was examined by the myosin heavy chain content of fibers by means of immunohistochemical staining and their cross‐sectional area was measured. All muscle fibers were identified as slow type I and fast type IIA, IIX or IIB (respectively, with increasing twitch contraction speed and fatigability). At lower activity levels (exceeding 5% of the peak activity), the duty time of the anterior belly of the digastric muscle was significantly higher in the soft‐diet group than in the hard‐diet group (P < 0.05). At higher activity levels (exceeding 20 and 50% of the peak activity), the duty time of the superficial masseter muscle in the soft‐diet group was significantly lower than that in the hard‐diet group (P < 0.05). There was no difference in the duty time of the anterior temporalis muscle at any muscle activity level. The percentage of type IIA fibers of the superficial masseter muscle in the soft‐diet group was significantly lower than that in the hard‐diet group (P < 0.01) and the opposite was true with regard to type IIB fibers (P < 0.05). The cross‐sectional area of type IIX and type IIB fibers of the superficial masseter muscle was significantly smaller in the soft‐diet group than in the hard‐diet group (P < 0.05). There was no difference in the muscle fiber composition and the cross‐sectional area of the anterior belly of the digastric and anterior temporalis muscles. In conclusion, for the jaw muscles of male rats reared on a soft diet, the slow‐to‐fast transition of muscle fiber was shown in only the superficial masseter muscle. Therefore, the reduction in the amount of powerful muscle contractions could be important for the slow‐to‐fast transition of the myosin heavy chain isoform in muscle fibers.

Effects of the masticatory demand on the rat mandibular development

The influence of masticatory loading stimulus on mandibular development is not fully clear. In this paper, experimental alterations in the daily muscle use, caused by a changed diet consistency, were continuously monitored, while adaptations in bone and cartilage were examined. It is hypothesised that decreased muscular loading will result in a decrease in the growth factor expression and mandible growth. Fourteen 21-day-old Wistar strain male rats were randomly divided into two groups and fed on either a hard or soft diet for 14 weeks. An implanted radio-telemetric device recorded continuously muscle activity of the superficial masseter muscle. Chondroblast proliferation in the condylar cartilage was identified by insulin-like growth factor-1 receptor (IGF-1r) immunostaining. Furthermore, an X-ray was taken for cephalometric analysis. In the soft-diet group, the duty time of the superficial masseter muscle at higher activity levels was significantly lower than that in the hard-diet group. This decrease in muscular loading of the jaw system was accompanied by: a significant reduction in (i) articular cartilage thickness, (ii) expression of IGF-1r immunopositive cells and (iii) mandible ramus height. In conclusion, a decrease in masticatory demand during the growth period leads to insufficient mandibular development.

Burst characteristics of daily jaw muscle activity in juvenile rabbits

SUMMARY Muscle activation varies with different behaviors and can be quantified by the level and duration of activity bursts. Jaw muscles undergo large anatomical changes during maturation, which are presumably associated with changes in daily muscle function. Our aim was to examine the daily burst number, burst length distribution and duty time (fraction of the day during which a muscle was active) of the jaw muscles of juvenile male rabbits (Oryctolagus cuniculus). A radio-telemetric device was implanted to record muscle activity continuously from the digastric, superficial and deep masseter, medial pterygoid and temporalis during maturation week 9-14. Daily burst characteristics and duty times were determined for activations, including both powerful and non-powerful motor behavior. All muscles showed constant burst numbers, mean burst lengths and duty times during the recording period. Including all behavior, the temporalis showed significantly larger daily burst numbers (205 000) and duty times (18.2%) than the superficial and deep masseter (90 000; 7.5%). Burst numbers and duty times were similar for the digastric (120 000; 11.1%) and medial pterygoid (115 000; 10.4%). The temporalis and deep masseter showed many short low activity bursts (0.05 s), the digastric showed many long bursts (0.09 s). For activations during powerful behaviors the superficial masseter and medial pterygoid had the largest burst numbers and duty times. Both muscles showed similar burst characteristics for all activation levels. It was concluded that activation of the jaw muscles is differently controlled during powerful and non-powerful motor behaviors and the functional organization of motor control patterns does not vary from 9 to 14 weeks of age.

Local applications of myostatin-siRNA with atelocollagen increase skeletal muscle mass and recovery of muscle function.

Background Growing evidence suggests that small-interfering RNA (siRNA) can promote gene silencing in mammalian cells without induction of interferon synthesis or nonspecific gene suppression. Recently, a number of highly specific siRNAs targeted against disease-causing or disease-promoting genes have been developed. In this study, we evaluate the effectiveness of atelocollagen (ATCOL)-mediated application of siRNA targeting myostatin (Mst), a negative regulator of skeletal muscle growth, into skeletal muscles of muscular dystrophy model mice. Methods and Findings We injected a nanoparticle complex containing myostatin-siRNA and ATCOL (Mst-siRNA/ATCOL) into the masseter muscles of mutant caveolin-3 transgenic (mCAV-3Tg) mice, an animal model for muscular dystrophy. Scrambled (scr) -siRNA/ATCOL complex was injected into the contralateral muscles as a control. Two weeks after injection, the masseter muscles were dissected for histometric analyses. To investigate changes in masseter muscle activity by local administration of Mst-siRNA/ATCOL complex, mouse masseter electromyography (EMG) was measured throughout the experimental period via telemetry. After local application of the Mst-siRNA/ATCOL complex, masseter muscles were enlarged, while no significant change was observed on the contralateral side. Histological analysis showed that myofibrils of masseter muscles treated with the Mst-siRNA/ATCOL complex were significantly larger than those of the control side. Real-time PCR analysis revealed a significant downregulation of Mst expression in the treated masseters of mCAV-3Tg mice. In addition, expression of myogenic transcription factors was upregulated in the Mst-siRNA-treated masseter muscle, while expression of adipogenic transcription factors was significantly downregulated. EMG results indicate that masseter muscle activity in mCAV-3Tg mice was increased by local administration of the Mst-siRNA/ATCOL complex. Conclusion These data suggest local administration of Mst-siRNA/ATCOL complex could lead to skeletal muscle hypertrophy and recovery of motor disability in mCAV-3Tg mice. Therefore, ATCOL-mediated application of siRNA is a potential tool for therapeutic use in muscular atrophy diseases.