Effect of pre-stress on the dynamic tensile behavior of the TMJ disc.

Previous dynamic analyses of the temporomandibular joint (TMJ) disc have not included a true preload, i.e., a step stress or strain beyond the initial tare load. However, due to the highly nonlinear stress-strain response of the TMJ disc, we hypothesized that the dynamic mechanical properties would greatly depend on the preload, which could then, in part, account for the large variation in the tensile stiffnesses reported for the TMJ disc in the literature. This study is the first to report the dynamic mechanical properties as a function of prestress. As hypothesized, the storage modulus (E') of the disc varied by a factor of 25 in the mediolateral direction and a factor of 200 in the anteroposterior direction, depending on the prestress. Multiple constant strain rate sweeps were extracted and superimposed via strain-rate frequency superposition (SRFS), which demonstrated that the strain rate amplitude and strain rate were both important factors in determining the TMJ disc material properties, which is an effect not typically seen with synthetic materials. The presented analysis demonstrated, for the first time, the applicability of viscoelastic models, previously applied to synthetic polymer materials, to a complex hierarchical biomaterial such as the TMJ disc, providing a uniquely comprehensive way to capture the viscoelastic response of biological materials. Finally, we emphasize that the use of a preload, preferably which falls within the linear region of the stress-strain curve, is critical to provide reproducible results for tensile analysis of musculoskeletal tissues. Therefore, we recommend that future dynamic mechanical analyses of the TMJ disc be performed at a controlled prestress corresponding to a strain range of 5­10%.

Regional dynamic tensile properties of the TMJ disc.

Although the TMJ disc has been well-characterized under tension and compression, dynamic viscoelastic regional and directional variations have heretofore not been investigated. We hypothesized that the intermediate zone under mediolateral tension would exhibit lower dynamic moduli compared with the other regions of the disc under either mediolateral or anteroposterior tension. Specimens were prepared from porcine discs (3 regions/direction), and dynamic tensile sweeps were performed at 1% strain over a frequency range of 0.1 to 100 rad/sec. Generally, the intermediate zone possessed the lowest storage and loss moduli, and the highest loss tangent. This study further accentuates the known distinct character of the intermediate zone by showing for the first time that these differences also extend to dynamic behavior, perhaps implicating the TMJ disc as a structure primarily exposed to predominantly anteroposterior tension via anterior and posterior attachments, with a need for great distension mediolaterally across the intermediate zone.

Enhanced crystallization of the Cys18 to Ser mutant of bovine gammaB crystallin.

The cysteine residues of the gamma crystallins, a family of ocular lens proteins, are involved in the aggregation and phase separation of these proteins. Both these phenomena are implicated in cataract formation. We have used bovine gammaB crystallin as a model system to study the role of the individual cysteine residues in the aggregation and phase separation of the gamma crystallins. Here, we compare the thermodynamic and kinetic behavior of the recombinant wild-type protein (WT) and the Cys18 to Ser (C18S) mutant. We find that the solubilities of the two proteins are similar. The kinetics of crystallization, however, are different. The WT crystallizes slowly enough for the metastable liquid-liquid coexistence to be easily observed. C18S, on the other hand, crystallizes rapidly; the metastable coexisting liquid phases of the pure mutant do not form. Nevertheless, the coexistence curve of C18S can be determined provided that crystallization is kinetically suppressed. In this way we found that the coexistence curve coincides with that of the WT. Despite the difference in the kinetics of crystallization, the two proteins were found to have the same crystal forms and almost identical X-ray structures. Our results demonstrate that even conservative point mutations can bring about dramatic changes in the kinetics of crystallization. The implications of our findings for cataract formation and protein crystallization are discussed.