Journal of Dental Research http://jdr.sagepub.

com/

Dynamic Properties of Bovine Temporomandibular Joint Disks Change with Age

E. Tanaka, J. Aoyama, M. Tanaka, H. Murata, T. Hamada and K. Tanne J DENT RES 2002 81: 618 DOI: 10.1177/154405910208100908 The online version of this article can be found at: http://jdr.sagepub.com/content/81/9/618

Published by:
http://www.sagepublications.com

On behalf of:
International and American Associations for Dental Research

Additional services and information for Journal of Dental Research can be found at: Email Alerts: http://jdr.sagepub.com/cgi/alerts Subscriptions: http://jdr.sagepub.com/subscriptions Reprints: http://www.sagepub.com/journalsReprints.nav Permissions: http://www.sagepub.com/journalsPermissions.nav

>> Version of Record - Sep 1, 2002 What is This?

Downloaded from jdr.sagepub.com at SRM DENTAL COLLEGE on December 14, 2012 For personal use only. No other uses without permission. International and American Associations for Dental Research

RESEARCH REPORTS
Biomaterials & Bioengineering

E. Tanaka*, J. Aoyama, M. Tanaka1, H. Murata2, T. Hamada2, and K. Tanne

Department of Orthodontics, Hiroshima University Faculty of Dentistry, 1-2-3 Kasumi, Minami-ku, Hiroshima 7348553, Japan; 1Division of Mechanical Science, Department of Systems and Human Science, Osaka University School of Engineering Science; 2 Department of Prosthetic Dentistry, Hiroshima University Faculty of Dentistry; *corresponding author, etanaka@hiroshima-u.ac.jp J Dent Res 81(9):618-622, 2002

Dynamic Properties of Bovine Temporomandibular Joint Disks Change with Age

ABSTRACT
The temporomandibular joint disk exhibits morphological and biochemical age-related changes. However, the possible age-related changes of the dynamic viscoelasticity in the disk are unclear. We tested the hypothesis that the dynamic viscoelastic properties of the disk change with age. Thirty-six disks from young-adult, adult, and mature-adult cattle were used for dynamic tensile tests. In all disks, the magnitudes of the complex modulus, the storage modulus, and the loss modulus increased as the frequency increased. The mature-adult disks had higher values of these moduli than did the younger disks. The loss tangent ranged from 0.1 to 0.3, which means that the disk has relatively large elasticity and relatively small viscosity. It was concluded that both the elasticity and viscosity of the disk increase with age. This may reflect age-related changes in biochemical composition. KEY WORDS: temporomandibular joint disk, dynamic viscoelasticity, aging.

INTRODUCTION

he disk of the temporomandibular joint (TMJ), located between the T temporal bone and the mandibular condyle, plays an important role as a stress absorber during function (Osborn, 1985; Tanne et al., 1991; Chin et al., 1996; Scapino et al., 1996; Kuboki et al., 1997; Tanaka et al., 1999). The TMJ disk consists mainly of type I collagen fibers and proteoglycans which are constrained in the interstices of the collagen fiber mesh (Scapino et al., 1996). Because of the composition and organization of its collagen fibers and glycosaminoglycans (GAGs), the TMJ disk exhibits viscoelastic characteristics (Nickel and McLachlan, 1994; Scapino et al., 1996; Kuboki et al., 1997; Tanaka et al., 1999). Studies of biomechanical responses to various loadings have provided useful information on the mechanical properties of various tissues and on their changes due to diseases or injuries (Oloyede et al., 1992; Iatridis et al., 1996; Ker et al., 2000; Tanaka et al ., 2000). Concerning the TMJ disk, several teststensile, compressive, creep, and stress relaxationhave provided valuable information on the viscoelastic properties, residual strain, and strain energy dissipation associated with time (Teng et al. , 1991; Scapino et al., 1996; Kuboki et al., 1997; Tanaka et al., 1999, 2002). These methods, however, are not sufficient for a rigorous evaluation of the rheological characteristics of the TMJ disk under a cyclic application of stress reflecting masticatory function (Beek et al., 2001). The automatic dynamic viscoelastometer, which is based on a principle of non-resonanceforced vibration, is a suitable instrument for evaluation of the viscoelastic properties of various materials (Clarke, 1989; Murata et al., 2000). The disk undergoes biochemical and morphological changes throughout life. These changes may be caused by changes of the loading condition in the TMJ. The calcium content of the disk increases progressively with aging (Takano et al., 1999); it has been suggested that the increase in calcifications is caused by aging or mechanical stress (Jibiki et al., 1999). Nakano and Scott (1996), using bovine TMJ disks, investigated the age-related changes in chemical composition and reported marked increases in total and sulfated GAGs in the inner tissues from mature fetuses to mature adults. Therefore, an age-related change in the viscoelastic properties can be expected. It is still unclear, however, whether the dynamic viscoelastic properties of the disk change with age. Such information may shed light on the age-related changes of the response of the TMJ disk to the applied mechanical stresses. In this in vitro study, we investigated the dynamic viscoelastic properties of the bovine TMJ disk over a wide range of frequencies. The aim was to evaluate the age-related effect on the biomechanical responses of TMJ disks, using a dynamic viscoelastometer.

MATERIALS & METHODS

Description of the Sample
Received September 7, 2001; Last revision June 14, 2002; Accepted July 8, 2002

Thirty-six bovine TMJ disks from 12 young-adult (3 yrs old), 12 adult (7 yrs old), and 12 mature-adult cattle (more than 10 yrs old) were obtained at a local

618

Downloaded from jdr.sagepub.com at SRM DENTAL COLLEGE on December 14, 2012 For personal use only. No other uses without permission. International and American Associations for Dental Research

J Dent Res 81(9) 2002

Dynamic Properties of Bovine TMJ Disks with Age

619

abattoir (Japan Agriculture, Hiroshima, Japan). The ages of the animals were estimated from the numbers of permanent anterior teeth. The protocol of the experiment was approved by the animal care and use committee at Hiroshima University. The disks were carefully dissected soon after the animals death. All disks were visually normal, oval, and had thicknesses and sizes within standard range (Meister and Berg, 1972). Immediately after resection, the disks were placed in 0.9% NaCl solution and stored at 4C; tensile tests were conducted within 8 hrs after resection. Each disk was sectioned antero-posteriorly, and a test specimen was derived from the central region (Fig. 1). To be able to determine the amount of applied stress (see below) for each specimen, we calculated the cross-sectional area by multiplying its thickness and its mediolateral width. Width and thickness were measured by means of digimatic calipers (CD-S20C, Mitutoyo Co., Kawasaki, Japan) and were 2.1 0.3 mm (mean SD) and 10.0 1.0 mm in the young-adult, 1.8 0.2 mm and 9.1 0.9 mm in the adult, and 1.8 0.1 mm and 9.6 0.3 mm in the matureadult groups; cross-sectional areas were 21.3 4.0, 16.3 2.5, and 17.3 1.1 mm2 (mean SD) in the young-adult, adult, and matureadult groups, respectively. The amount of antero-posterior length of the specimens was approximately 14 mm, including the length (about 4 mm) for gripping of the specimen in the dynamic viscoelastometer (see below).

the viscous component of the material behavior. The former is directly proportional to the energy storage in a cycle of deformation, and the latter is proportional to the average dissipation or loss of energy. The loss tangent tan is the ratio of energy lost to energy stored during cyclic deformation. The magnitude of complex modulus E* was determined by E* = / Based on the phase angle , the storage and loss moduli, E and E, were determined by E* = E + iE E = E* cos E = E* sin __ where i = 1 and tan = E/E is the loss tangent. The dependence of the dynamic viscoelasticity on age was statistically evaluated by use of the results at 1.0 Hz. This frequency reflects masticatory conditions (Druzinsky, 1993; Gallo et al ., 2000). We performed Students t tests to test for the differences among mean values of E, E, and tan among the three groups of specimens.

RESULTS
In all age groups, the values of E*, E, and E increased as the frequency increased from 0.1 to 100 Hz (Fig. 2). Note that the specimens from the young-adult group showed a relatively small change with frequency. The mature-adult group had higher values of E*, E, and E than did the younger groups, regardless of frequency. In all groups, the loss tangent exhibited relatively small changes between 0.1 and 1 Hz; it increased with frequencies beyond 1 Hz (Fig. 3). Throughout the entire range of frequencies tested, the mature-adult group had a larger loss tangent than did the younger groups. The young-adult and adult groups showed almost the same loss tangent, regardless of frequency. The tan ranged from 0.1 to 0.3, which means that the TMJ disk has a relatively large elasticity and a relatively small, but not negligible, viscosity.

Testing Apparatus and Conditions

The dynamic viscoelastic properties of the specimens were determined with the use of an automatic dynamic viscoelastometer (Rheovibron DDV-25FP, Orientec Corp., Tokyo, Japan) (Fig. 1). This device consists of a measurement operation block, a main unit, a power unit, a data-processing device, and testing jigs. The measurement operation block consists of a magnetic exciter, a tension control motor, an amplitude-detecting sensor and an amplifier, a loaddetecting load cell, and chucks. The specimen was rigidly clamped in the two chucks; the chuck-to-chuck distance was 10 mm. All experiments were performed in a bath with 0.9% NaCl solution at room temperature. During the test, a dynamic tension was applied to the specimen along an antero-posterior axis by a sinusoidal strain of = 0 + sin(t), with an administrative strain of 0 = 0.5% and an oscillation amplitude of 2 = 0.2%. The stress behavior was described by = 0 + sin(t + ). The sinusoidal strain was produced by the tension control motor in the driver; the time-dependent stress and strain were measured by means of load and displacement detectors, respectively. The main unit transmits input-output signals to the measurement operation block and data processor. The measurement operation unit detected the shape-error of the sinusoidal strain and the slippage between chuck and specimen and adjusted them automatically. In the present study, the oscillation frequency ranged from 0.1 to 100 Hz.

Dynamic Viscoelastic Parameters

Due to the viscosity, the stress-strain response is generally out of phase, and the phase difference between the stress and strain is somewhere between 0 and 90 degrees (Fig. 1). Based on the dynamic behavior of stress and strain, the complex tensile modulus E*, the tensile storage modulus E, the tensile loss modulus E, and the loss tangent tan were determined as dynamic viscoelastic parameters. The complex modulus E* can be resolved into E and E. The storage modulus (E) is the ratio of the stress in phase with the strain to the strain and represents the elastic component of the material behavior. The loss modulus (E) is the ratio of the stress 90 degrees out of phase with the strain to the strain and represents
Figure 1. Block diagram of the dynamic viscoelastometer with a schematic representation of the relationship between stress and strain of a viscoelastic material (A), and an illustration showing the anatomy of the bovine TMJ disc and the location of the specimen dissected (B). The sinusoidal strain is produced by a tension control motor in the driver, and the stress and strain are measured by means of load and displacement detectors and transmitted to a data processor. In a viscoelastic material, the phase difference between stress and strain is somewhere between (/2 > > 0), and the complex modulus E* is resolved into two components, i.e., the storage modulus E and the loss modulus E, shown vectorially. Furthermore, the tangent of the phase angle () between stress and strain is a measure of the ratio of energy loss to energy stored during cyclic deformation.

Downloaded from jdr.sagepub.com at SRM DENTAL COLLEGE on December 14, 2012 For personal use only. No other uses without permission. International and American Associations for Dental Research

620

Tanaka et al.

J Dent Res 81(9) 2002

larger than E in the younger groups (p < 0.01). With respect to the loss tangent tan , the mature-adult group had the highest value (0.16 0.03); this value was significantly larger than that in the adult group (p < 0.01). The value of tan in the adult group was almost the same as that in the young-adult group.

DISCUSSION
The TMJ disk is subjected to intermittent loading during mastication and to sustained loading during clenching. In response to dynamic force conditions, the disk behaves in a viscoelastic fashion, which is different from its behavior during static conditions. It was found that the dynamic viscoelastic behavior of the bovine TMJ disks is sensitive to changes in frequency regardless Figure 2. The complex modulus E*, storage modulus E, and loss modulus E as a function of frequency in the young-adult (A), the adult (B), and the mature-adult groups (C). Error bars are standard deviations of age. All of the dynamic (for each group, n = 12). The frequency has a proportional effect on the values of the moduli. Complex viscoelastic parameters became modulus E*; x Storage modulus E ; and l Loss modulus E . larger with an increase of frequency. This implies that, at higher frequencies, a larger load is required to produce the same strain. Thus, at higher frequencies, the disk is better able to resist higher loads than at lower frequencies. This property may come from the wavy structure of the collagen fibers comprising the TMJ disk under unstrained condition. Collagen crimping in the human TMJ disk is observed at ages ranging from 4.5 to 63 yrs (Berkovitz, 2000). It should be realized that, in the current experiment, more than 10 cycles were applied for each frequency and that the measured moduli were based on a steady-state response. Under cyclic loading, a steady-state response of the disk, as in other viscoelastic tissues, is usually reached within 10 cycles (see also Beek et al. , 2001). Therefore, it is not likely that the increase in the moduli with frequency observed in the present study is due to a time-dependent effect. To rule out such an effect, we carried out experiments (for each age group, n = 5) in which the moduli were also measured at several selected constant frequencies (1, 10, and 100 Hz) with increasing cycles (data not shown). Except for the above-mentioned initial Figure 3. Loss tangent vs. frequency. Error bars are standard deviations. response to reach a steady state, there was no effect of cycle Young-adult, p Adult, and l Mature-adult groups. number, and the steady-state moduli at these constant frequencies did not differ significantly from those obtained The storage modulus E, loss modulus E, and loss tangent with the regular experimental protocol. tan at 1.0 Hz differed among the groups (Fig. 4). The storage In the present study, the total strain was 0.5%. Within this modulus E was 0.69 0.21 MPa (mean SD) in the youngrange of strain, the predominant increase in the resultant stress adult group, 1.21 0.41 MPa in the adult group, and 1.44 with an increase of frequency may be related to the movement 0.31 MPa in the mature-adult group; the storage modulus of the of water rather than to the resilience of the collagenous portion young-adult group was significantly smaller than that of the of the matrix. Below a certain level of frequency, the older groups (p < 0.01). In the mature-adult group, the loss morphological change of the TMJ disk may match up to the modulus E was 0.23 0.06 MPa, which was significantly water movement within the tissue. At higher strain rates (higher

Downloaded from jdr.sagepub.com at SRM DENTAL COLLEGE on December 14, 2012 For personal use only. No other uses without permission. International and American Associations for Dental Research

J Dent Res 81(9) 2002

Dynamic Properties of Bovine TMJ Disks with Age

621

frequency), the frictional resistance due to interstitial fluid flow relative to the solid matrix (collagen matrix) may increase, resulting in a higher stiffness; the dissipation due to interstitial fluid flow may also increase, resulting in a larger hysteresis loop with an increase of frequency. In the present study, the storage modulus E of the disk was less than 2.5 MPa, regardless of frequency and age, which is smaller than the elastic moduli previously measured under static conditions (Tanne et al., 1991; Teng et al., 1991; Tanaka et al., 1999). In general, the dynamic properties are determined in cycle tests at low strain levels, as was the case in the study. Furthermore, it is known that the curves showing the experimental peak-and-valley stresses in cyclic testing match well with those obtained from the theoretical stress-relaxation curves according to the quasi -linear viscoelastic theory (Woo, 1986). Recently, we investigated the stress relaxation property of bovine TMJ disks in compression and found that the relaxed modulus ranged from 1.1 to 2.3 MPa (del Pozo et al. , 2002). The values of the relaxed modulus have some resemblance to the dynamic viscoelastic parameters in the present study. This finding implies that, under dynamic conditions, the disk exhibits a biomechanical equilibrium which is similar to that after stress-relaxation. In this study, we examined a wide range of frequencies. This range is relatively large compared with the upper limit of the chewing frequency in the cow (Druzinsky, 1993). It should be realized, however, that, during mastication, impulse loadings may occur which include high-frequency components. Within the wide frequency range examined in the present study, values of 0.5 to 1.0 Hz reflect masticatory conditions (Druzinsky, 1993; Gallo et al. , 2000). These values are considered to be important for assessment of the clinical significance of the results. Therefore, in this study, the dependence of dynamic viscoelasticity on aging was evaluated by use of the results at 1.0 Hz. At this frequency, the mature-adult group had significantly higher values than the young-adult group in storage modulus E, loss modulus E, and loss tangent tan. The storage modulus E describes elastic deformation under loading, while the loss modulus E represents viscous deformations. Therefore, it can be concluded from the present study that both the elasticity and viscosity of the TMJ disk increase with age. The present finding that the values of the viscoelastic parameters of the bovine TMJ disk increase with age is consistent with the findings of previous biochemical studies (Nakano and Scott, 1996; Nishimura et al., 1998). The increase in loss tangent implies an increase of viscosity relative to elasticity. Functionally, the damping, which results from a higher value of loss tangent, is likely to produce a degree of stress relief under masticatory forces. Therefore, the matureadult disk may have a greater capacity for stress distribution and thus may reduce stress concentration at the surfaces of the articulating bones.

Figure 4. Means and standard deviations of storage modulus E and loss modulus E (A), and loss tangent tan (B) at 1.0 Hz. Asterisks: significance of differences among the groups (** p < 0.01) as tested with Students t test. The mature-adult group had significantly larger moduli than the younger groups.

REFERENCES
Beek M, Aarnts MP, Koolstra JH, Feilzer AJ, van Eijden TM (2001). Dynamic properties of the human temporomandibular joint disc. J Dent Res 80:876-880. Berkovitz BK (2000). Collagen crimping in the intra-articular disc and articular surfaces of the human temporomandibular joint. Arch Oral Biol 45:749-756. Chin LP, Aker FD, Zarrinnia K (1996). The viscoelastic properties of the human temporomandibular joint disc. J Oral Maxillofac Surg 54:315-318. Clarke RL (1989). Dynamic mechanical thermal analysis of dental polymers. 1. Heat-cured poly(methyl methacrylate)-based materials. Biomaterials 10:494-498. del Pozo R, Tanaka E, Tanaka M, Okazaki M, Tanne K (2002). The regional difference of viscoelastic property of bovine temporomandibular joint disc in compressive stress-relaxation. Med Eng Phys 24:165-171. Druzinsky RE (1993). The time allometry of mammalian chewing movements: chewing frequency scales with body mass in mammals. J Theoret Biol 160:427-440. Gallo LM, Nickel JC, Iwasaki LR, Palla S (2000). Stress-field translation in the healthy human temporomandibular joint. J Dent Res 79:1740-1746. Iatridis JC, Weidenbaum M, Setton LA, Mow VC (1996). Is the nucleus pulposus a solid or a fluid? Mechanical behaviors of the nucleus pulposus of the human intervertebral disc. Spine 21:1174-1184. Jibiki M, Shimoda S, Nakagawa Y, Kawasaki K, Asada K, Ishibashi K (1999). Calcifications of the disc of the temporomandibular joint. J Oral Pathol Med 28:413-419. Ker RF, Wang XT, Pike AV (2000). Fatigue quality of mammalian tendons. J Exp Biol 203:1317-1327. Kuboki T, Shinoda M, Orsini MG, Yamashita A (1997). Viscoelastic properties of the pig temporomandibular joint articular soft tissues of the condyle and disc. J Dent Res 76:1760-1769. Meister R, Berg R (1972). Topographical and applied anatomy of the temporomandibular joint of various domestic animals with special reference to resection of the articular disc. 2. Calf (Bos taurus). Z Exp Chir 5:271-282. Murata H, Taguchi N, Hamada T, McCabe JF (2000). Dynamic viscoelastic properties and the age changes of long-term soft denture liners. Biomaterials 21:1421-1427. Nakano T, Scott PG (1996). Changes in the chemical composition of

ACKNOWLEDGMENTS
This research was supported in part by a grant (No. 12771280) for Science Research from the Ministry of Education, Science and Culture, Japan.

Downloaded from jdr.sagepub.com at SRM DENTAL COLLEGE on December 14, 2012 For personal use only. No other uses without permission. International and American Associations for Dental Research

622

Tanaka et al.

J Dent Res 81(9) 2002

the bovine temporomandibular joint disc with age. Arch Oral Biol 41:845-853. Nickel JC, McLachlan KR (1994). In vitro measurement of the stressdistribution properties of the pig temporomandibular joint disc. Arch Oral Biol 39:439-448. Nishimura M, Tan W, Mukudai Y, Nakamura S, Nakamasu K, Kawata M, et al . (1998). Role of chondroitin sulfate-hyaluronan interactions in the viscoelastic properties of extracellular matrices and fluids. Biochim Biophys Acta 1380:1-9. Oloyede A, Flachsmann R, Broom ND (1992). The dramatic influence of loading velocity on the compressive response of articular cartilage. Connect Tissue Res 27:211-224. Osborn JW (1985). The disc of the human temporomandibular joint: design, function and failure. J Oral Rehabil 12:279-293. Scapino RP, Canham PB, Finlay HM, Mills DK (1996). The behaviour of collagen fibres in stress relaxation and stress distribution in the jaw-joint disc of rabbits. Arch Oral Biol 41:1039-1052. Takano Y, Moriwake Y, Tohno Y, Minami T, Tohno S, Utsumi M, et al . (1999). Age-related changes of elements in the human articular disk of the temporomandibular joint. Biol Trace Elem Res 67:269-276.

Tanaka E, Tanaka M, Miyawaki Y, Tanne K (1999). Viscoelastic properties of canine temporomandibular joint disc in compressive load-relaxation. Arch Oral Biol 44:1021-1026. Tanaka E, Miyawaki Y, del Pozo R, Tanne K (2000). Changes in biomechanical properties of the rat interparietal suture incident to continuous tensile force application. Arch Oral Biol 45:1059-1064. Tanaka E, Tanaka M, Aoyama J, Watanabe M, Hattori Y, Asai D, et al. (2002). Viscoelastic properties and residual strain in a tensile creep test on bovine temporomandibular articular disks. Arch Oral Biol 47:139-146. Tanne K, Tanaka E, Sakuda M (1991). The elastic modulus of the temporomandibular joint disc from adult dogs. J Dent Res 70:1545-1548. Teng S, Xu Y, Cheng M, Li Y (1991). Biomechanical properties and collagen fiber orientation of temporomandibular joint discs in dogs: 2. Tensile mechanical properties of the discs. J Craniomandib Disord 5:107-114. Woo SL-Y (1986). Biomechanics of tendons and ligaments. In: Frontiers in biomechanics. Schmid-Schonbein GW, Woo SL-Y, Zweifach BW, editors. New York: Springer-Verlag, Chapter 14, pp. 180-195.

Downloaded from jdr.sagepub.com at SRM DENTAL COLLEGE on December 14, 2012 For personal use only. No other uses without permission. International and American Associations for Dental Research