In vitro study on biomechanical characteristics of tendons of limbs of the buffalo* S.C.dubal, 1 K.N.Vyas 2 and Y.L.

Vyas 3 Department of Anatomy College of Veterinary Sc. & A.H., Gujarat Agricultural University, Anand 388 001 (Guj) India Key words: Biomechanical characteristics, tendons, limbs, buffalo. ABSTRACT The biomechanical characteristics of tendons of distal part of the fore and hind limbs of six young (6 to 10 months old) and adult (5 to 10 years old) male Surti buffaloes were utilized to understand quantitatively the susceptibility of the tendons for strain injuries. There were significant (P < 0.01) age related differences amongst the tendons studied. The mean value of Young` s modulus of elasticity, factor of safety and fracture toughness did not differ (P >0.01) amongst the tendons studied, and hence these properties appeared to be material properties. All other mechanical properties appeared to be specimen properties. Those tendons which act as prime movers and are subjected to high physical stresses, had higher values of tensile strength but lower values of critical length of micro-damage and the number of fatigue cycles than their opponents. It was therefore concluded that former tendons were more susceptible to tendon injuries than the later ones. INTRODUCTION The aetiology of tendon injuries in domestic animals is, in general, rather complicated. It is an emerging concept in musculo-skeletal pathology that cyclic over loading creates cumulative micro-damage which ultimately results in tissue failure. Most of the biomechanical studies have been carried out on equine tendons. The suggested factors responsible for micro-damage of equine tendons are cross-sectional area and collagen content (Riemersma and Schamhardt, 1985), composition of extra-cellular matrix (Jones and Boe, 1990), longitudinal heterogenecity (Smith et al.,1994), inter fibre differences (Becker et al,. 1994) and elevation of core temperature ( Wilson and Goodship,1994 ). There are a few reports pertaining to the biomechanical studies of the bovine tendons (Pandya, 1982, Vyas and Vyas, 1985). However, these studies did not concern with the tendon injuries in general and micro-damage and cyclic over loading in particular. The present work was an attempt to _______________________________________

Part of Ph.D. thesis submitted by first author.

1

Assistant professor, 2 Retd. Principal and 3Associate professor.

compare the biomechanical properties of tendons of distal part of both the limbs of the buffalo so that the study may be useful in understanding the tendon injuries. Emphasis was especially so that the study may be useful in understanding the tendon injuries. Emphasis was especially given to the transverse length of micro-damage and number of cycles, which can cause the tendon failure. MATERIALS AND METHODS Fresh muscles with their tendons were obtained from cadaver fore and hind limbs of apparently healthy six male young (6 to 10 months old) and adult ( 5 to 10 years old ) Surti buffaloes. The mean body weight of young and adult animals were 64.54 6.74 kg and 518.84 14.55 kg respectively. The tendons were immediately kept in ice cooled normal saline solution. The widest part of muscle belly was cut transversely then its impression was recorded on a graph paper and subsequently its cross-sectional area (CSA in cm2) was measured. The maximum force generated by the muscle was termed as working load (WL in kg).The working load was calculated by multiplying the CSA of the muscle belly with 6 i.e., WL = 6 CSA ( Ikai and Fukunaga, 1968). From the thinnest part of the tendon, two perpendicular diameters were taken. The CSA was almost elliptical in shape. The tendons were then immediately subjected to study the tensile loading at testing machine. For each tendon, a stress-strain curve was obtained to measure the young` s modulus of elasticity (E). The ultimate tensile load (UL) at which the tendon ruptured was also recorded. During the experiment the tendons were kept cool and moist by applying the cold normal saline solution. Tensile strength (TS) of a tendon was obtained by dividing the UL with its original CSA. Factor of safety (FS) was obtained by dividing the UL with the WL. The critical length (CL) of destructive process (the transverse length of micro-damage) was calculated as follows:

CL G

= =

2 G E / (TS) 2, Surface free energy of the tendon per unit area

(1)

The exact value of G is not known. The most probable value of G was assumed to be 0.50 10 3 J / m2 ( Pascoe 1978 and Epifanov 1979 ). The fracture toughness (FT) was calculated as follows: FT = = (TS) ( CL) (2 G E) (2)

The fatigue tensile strength (FTS) of a tendon was obtained by its minimum WL (assumed to be equal to 4 CSA of its belly. The number of cycles (N) of repeated loading that could produced the failure, was calculated as follows (Pascoe 1978 ): (d CL/dN) C = = C ( FTS ) n Constant. ( CL ) n/2 (3)

Since CL<< diameter of the tendon, then for n = 2, from eq (3) we get, N = (CL) / C ( FTS ) 2 (4)

The significance of mean differences amongst tendons was obtained by applying the multiple Duncan` s `t` test. The significance of age difference was obtained by applying the Student ` s pair test` t`. RESULTS The results of the present study are summarized in tables 1, 2, 3 and 4. Biomechanical properties in relation to age: The mean values of factor of safety and tensile strength of tendons of young animals were significantly (P < 0.01) more than those of the adult ones. However, the working load, ultimate load, Young ` s modulus of elasticity, critical length of micro-damage, number of

cycles of fatigue failure, fracture toughness and fatigue tensile strength showed significantly (P< 0.01) higher mean values in the adult animals than the young one. Biomechanical properties in relation to limbs and tendons: 1. Working load and Ultimate load: In both the age groups, the mean values of WL and UL of the tendons of hindlimb were significantly (P< 0.05) higher than those of the forelimb. Further, the WL and UL of tendons of cranio-lateral (tarsal flexors and digital extensors) group showed significantly (P< 0.05) lower mean values than those of the caudo-medial (tarsal extensors and digital flexors ) group in both the limbs ( Table 1). 2. Tensile strength and Fatigue tensile stress: The mean values of TS and FTS of the tendons of forelimb, were significantly (P< 0.05) more than those of the hind limb. Further, the tendons of cranio-lateral group of both the limbs had significantly (P<0.05) higher mean values of TS and FTS than those of the caudomedial group (Table 2). 3. Young ` s modulus of elasticity, Fracture toughness and Factor of safety: The mean values of E, FT and FS did not differ significantly (P> 0.01) amongst tendons studies (Table 3). The pooled mean values were 9.50 o.25 kN / mm 2, 173. 95 2.47 N/mm 3/2 and 5 0.70, respectively, in the calves and 19.50 0.30 kN / mm2, 330.58 27.06 N / mm 3/2 and 3 0.20, respectively, in the adult buffaloes. 4. Critical length of micro-damage, Number of fatigue cycles: On an average the tendons of cranio-lateral group of the forelimb showed lower mean values of CL and N than those of the caudo-medial group (Table 4). However, an opposite trend was observed in the tendons of the hind limb. Further, the digital extensor tendons of the forelimb had significantly (P< 0.05) lower mean values of N than those of the hind limb.

Further more, the superficial digital flexor tendon (SDFT) of both limbs had significantly (P< 0.05) lower mean values of CL than the deep digital flexor tendon (DDFT). Though the Achilles tendon showed the highest value of the WL and UL, the mean value of TS and FS were comparable with that of other tendons (Table 1,2 and 3). The flexor tendons of carpal and tarsal joints showed high value of the CL and N. DISCUSSION The results of this in vitro study can be discussed at two levels (1) from a biomechanical and (2) with more caution, from clinical view point. The factor of safety is the measure of design of safety margin of working load. The tendon is one of the important components of locomotion machine. Therefore the tendon should work below its maximum strength. Hence, the FS of tendons studied was greater than the unity (Table 3). The Young `s modulus of elasticity indicates the internal strength of a material against the external forces applied (Case and Chivler, 1971). The ultimate load when expressed in terms of unit area is known as tensile strength. Theoretical value of TS should about 0.1 E. However, the observed TS is always several times less (Table - 2 and 3). This discrepancy is due to the presence of micro-cracks which reduces the strength (Epifanov, 1979). According to Guy (1976), there is initially a tiny internal void, which grows into a micro crack in tensile strain. Further formation of a crack relieves the elastic stress. As long as the length of the micro-crack remains below a certain value, energy is required for it to develop. Further extension of tendon results in a reduction of its energy. Thus, a rise in temperature will enhance the progress of micro-crack. The cyclic tensile loading of equine tendon has been shown to result in an elevation of core temperature (Wilson and Goodship, 1994). The cyclic overloading creates cumulative micro-damage. When the transverse length of this micro-damage reaches a critical value (say critical length of destructive process or simply critical length of micro-damage), there is spontaneous rupture of the material. An

increase in tensile strength decreases in the plastic work done in initiating fracture at the tip of flaws. The rate of growth of micro-crack is related with a measurement known as stress concentration factor (K). It describes the distribution of stresses at the crack tip. When the value of K achieves a critical value, it is known as fracture toughness, there is catastrophic failure of the material (Pascoe, 1978). The extensor tendons of fore limb, the gastrocnemius tendon and Achilles tendon had high tensile strength. Hence, these tendons appeared to be highly susceptible to develop flaws. A low but repeated (continuous or intermittent) stress can cause the rupture of the tendon. This minimum stress is known as fatigue tensile stress. Which is obtained by stressstrain (SN) curve. Since such study was not conducted, the FTS was assumed to be minimum working load (= 4 CSA of muscle belly) per unit CSA of tendon. The calculated number of cycles gave an understanding of tendon failure. The mean values of E, FS and FT did not differ significantly (P>0.01) amongst the tendons studied. These biomechanical properties appeared to be material properties. This inference is in resonance with the reports of Wilmink et al. (1992). According to them the (Young` s) elastic modulus of collagen fibres obtained from SDFT of the fore limb of the race horse, did not vary significantly between the peripheral and core fibres or young and old horses. However, the present findings of significant age differences in mean values of these measurements, could be attributed to the differences in the tendon materials as the result of on going maturation processes in both the matrix and the fibres. The mean values of all other biomechanical measurements differed significantly amongst the tendons studied. The biomechanical properties appeared to be speciemen properties. This could be described to differential growth of the muscles and their tendons. This type of growth is functionally oriented ( Hildebrand, 1974). The present findings of WL, UL, FS and TS of the digital tendons are in close

agreement with the reports of Vyas and Vyas (1985). However, the TS of Achilles tendon of human being as reported by Mason and Allen (1947), and the UL and TS of accessory ligament of DDFT of equine fore limb, as reported by Becker et al. (1994), were higher than those of the present findings. These differences could be due to the differences in the specimen. From clinical point of view, the critical length of micro-damage and the number of cycles of fatigue failure are important. The constant (C), appeared in eq (3), is a material property (Guy, 1976). Since the value of C was not known, the value of N was expressed in multiple of C-1 (Table 4). The critical length of micro-damage indicates the maximum transverse damage, which a tendon can sustain under tensile strain. When the tendon attains the critical length, it ruptures under the applied stress. The higher the stress the less is the durability of the tendon. The tendons of forelimbs could sustain smaller damage and less number of cyclic loading compared to the tendons of hind limb. The former tendons were therefore more liable to tendon injuries than the later ones. This interpretation is in agreement with that of Stashak (1987) in the horses. Further, in the forelimbs, the extensor tendons could sustain smaller micro-damage and cycles of fatigue failure than the flexor tendons. Hence, the extensor tendons seemed to be more prone to tendon injuries than the flexor tendons. Though Stashak (1987) mentioned that the flexor tendon of fore limb of race horses and hunters are more liable to tendon injuries than the extensor tendons. The tendon injuries are directly related with the excess physical stress. The above discrepancy could be ascribed to differences in work stresses imposed on the buffaloes and the racehorses. In case of the hind limb, the extensor tendons of the tarsal joint and the digital flexor tendons (DFT) could sustain larger damage but low fatigue cycles than their opponent tendons. The tendon of gastrocnemius muscle and Achilles tendon appeared to be the most injury prone tendons. Krishnamurthy (1993) has also stated the same in the ruminants. Further

more, the DFT of the hind limb appeared to be more liable to the tendon injuries than those of the forelimb. This could be attributed to the differences in the functional status of these tendons. The DFT of the hind limb propel the whole body, hence, they are always subjected to high tensile stress. The DFT of the forelimb balances the body during motion, they are therefore subjected to relatively lower tensile stress. Within the forelimb, the SDFT could sustain shorter micro-damage and fatigue cycles than the DDFT. Therefore, the SDGT should be more liable to tendon injuries than the DDFT. This interpretation is in agreement with that of Stashak (1987) in the horses. However, according to Greenough et al. (1972), in cattle, no particular tendon has been recorded as being more liable to strain injuries than others. From this study it was concluded that the biomechanical characteristics like Young ` s modulus of elasticity, factor of safety and fracture toughness, were material properties. All other mechanical properties studied were specimen properties. There were age-related changes in the biomechanical properties of the tendons. Those tendons, which are prime movers and / or are subjected to high tensile stress, had higher tensile strength and were more susceptible to tendon injuries than their opponent tendons.

REFERENCES Becker, C. K, Sandberg, H. H C. M. and Barneveld, A 1994. In vitro mechanical properties of the accessory ligament of the deep digital flexor tendons in horses in relation to age.Equine Veterinary Journal, 26: 454 459. Case, J. and Chilver, A. H.1971. Strength of materials and Structures. 2nd edn. E.L.B.S and Edward Arnold Ltd,, London. Epifanov, G. I.. 1979. Solid state physics .Mir Publishers, Moscow. Greenough, P. R., MacCallum, F. J. and Weaver, A. D. 1972. Lameness in cattle. Cline and Boyd, Edinburgh.

Guy, A.G.1976. Essentials of material science. McGraw-Hill Kogakusha Ltd., New York. Hildebrand, M.1974. Analysis of vertebrate structures. John Willey and Sons, New York. Ikai, M. and Fukunaga, T.1968. Calculation of muscle strength per unit cross-sectional area of humane muscle by ultrasonic measurements. . Int. Z. Angew Physiol. Einschl Cinchiesslich Arbcitsphysiologia, 26:26 Jone, A. J. and Boe, J. A. 1990. Age and position related heterogenecity of equine tendon extra-cellular matrix composition. Research in Veterinary Science, 48: 357 364. Krishnamurthy, D. 1993. Tendons and ligaments. In : Ruminant Surgery. Eds. R.P.S. Tyagi and Jit Singh, CBS Publishersand Distributors, Delhi. Mason, M. L. and Allen, M. S. 1947. The rate of healing of tendons. An experimental study of tensile strength. Annals of Surgery.113: 424 450 Pandya, S. T. 1982. An experimental study on healing of sectional tendons in the bovine. M.V.Sc. Thesis, Gujarat Agricultural University, Anand. Pascoe, K. J. 1978. An Introduction to the properties of engineering material.3rd edn. E.L.B.S. and Van Nostrand Reinhold Co., London. Riemersma, D. J. and Schamhardt, H. C.1985. In vitro mechanical properties of equine tendons in relation to cross-sectional area and collagen content. Research in Veterinary Science. 39: 263 270. Smith, R. K. W, Jones, R. and Webbon, P. M.1994. The cross-sectional area of normal equine digital flexor tendons determined ultra-sonograhpically. Equine Veterinary Journal. 26: 460 465. Stashak, T. S. 1987. Adam` s Lameness in Horse. Ed.T.S. Stashak. Lea and Febiger, Philadelphia Vyas, Y. L. and Vyas, K. N. 1985. Tensile strength and factor of safety of the digital tendons in Surti buffalo and Kankrej bullock. J Anim. Sci. 56:1017 1022.

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Wilmink, J. Wilson, A. M. and Goodship, A. E. 1992. Functional siginificance of morphology and micro-mechanics of partial rupture of the SDFT in race horse. Research in Veterinary Science. 53: 345 359. Wilson, A. M. and Goodship, A. E. 1994. Exercise induce hyperthermia as a possible mechanism for tendon degeneration. J. Biomech. 27: 899 905.

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Table 1: Working load (WL, kN) and ultimate load (UL, kN) of tendons of buffalo. ________________________________________________________________________ Name of muscle (tendon) Buffalo calf Adult buffalo ( No. of animals = 6) WL UL WL UL _______________________________________________________________________ I. Forelimb: A. Cranio-lateral group: 1. Extensor carpii radialis 0.20 0.01 1.18 0.01 0.75 0.08 2.82 0.20 2. Medial digital extensor 0.28 0.01 0.70 0.05 0.52 0.02 0.92 0.10 3. Common digital extensor 0.35 0.02 0.74 0.03 0.31 0.01 0.72 0.06 4. Lateral digital extensor 0.10 0.01 0.86 0.08 0.61 0.02 1.04 0.10 B. Caudo-medial group: 1. Flexor carpii radialis 0.20 0.01 0.47 0.04 0.37 0.01 1.36 0.11 2. Flexor carpii ulnaris 0.21 0.01 0.60 0.03 0.41 0.02 1.02 0.05 3. Ulnaris lateralis 0.39 0.04 0.88 0.16 0.76 0.03 1.72 0.10 4. superficial digital flexor 0.21 0.02 1.02 0.13 0.61 0.03 1.74 0.13 5. Deep digital flexor 0.30 0.03 1.80 0.09 0.63 0.04 1.90 0.20 II. Hind limb A. Cranio-lateral group: 1. Peroneus tertius 0.09 0.01 1.11 0.09 0.56 0.03 1.62 0.12 2. Peroneus longus 0.17 0.01 0.65 0.09 0.75 0.04 2.06 0.10 3. tibialis cranialis 0.10 0.01 0.69 0.05 0.51 0.02 1.31 0.09 4. Medial digital extensor 0.27 0.02 0.60 0.08 0.41 0.03 1.28 0.07 5. Common digital extensor 0.40 0.04 0.76 0.07 0.57 0.03 1.50 0.02 6. Lateral digital extensor 0.18 0.01 0.60 0.03 0.37 0.02 1.10 0.10 B. Caudo-medial group: 1. Gastrocnemius 0.15 0.01 1.00 0.9 1.15 0.08 2.15 0.17 2. Achilles tendon 0.51 0.04 4.34 0.25 1.83 0.04 7.80 0.24 3. Superficial digital flexor 0.22 0.03 1.80 0.16 0.65 0.04 2.02 0.14 4. Deep digital flexor 0.72 0.05 1.95 0.18 1.13 0.10 2.04 0.18 The mean values of the measurements differed significantly between the age (P<0.01) and the limb groups (P<0.05).

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Table 2: Tensile strength (TS, N/ sq mm) and fatigue tensile stress (FTS, N /sq mm ) of tendons of the buffalo. _________________________________________________________________________ Name of muscle (tendon) Buffalo calf Adult buffalo ( No. of animals = 6) TS FTS TS FTS I. Forelimb: A. Cranio-lateral group: 1. Extensor carpii radialis 40.68 1.50 07.17 0.06 31.25 1.25 09.00 0.10 2. Medial digital extensor 96.15 1.89 40.00 1.50 35.27 1.20 20.00 0.20 3. Common digital extensor 105.0 2.05 90.00 1.20 40.10 1.30 17.10 0.20 4. Lateral digital extensor 84.70 1.50 10.79 1.00 24.00 1.20 15.01 0.20 B. Caudo-medial group: 1. Flexor carpii radialis 36.20 1.40 15.80 1.00 25.06 1.2 07.40 0.10 2. Flexor carpii ulnaris 55.12 1.50 20.25 1.00 26.75 1.10 08.75 0.10 3. Ulnaris lateralis 70.25 1.40 30.55 1.20 32.20 1.40 14.64 0.20 4. Superficial digital flexor 53.65 1.60 11.45 1.00 26.14 1.30 09.10 0.10 5. Deep digital flexor 50.46 1.50 09.86 1.00 25.05 1.20 09.00 0.10 II. Hind limb A. Cranio-lateral group: 1. Peroneus tertius 44.40 1.30 0.40 0.05 27.22 1.20 09.72 0.01 2. Peroneus longus 36.58 1.37 10.00 0.08 42.00 1.30 15.24 0.01 3. Tibialis cranialis 39.10 1.20 05.60 0.04 23.92 1.20 09.85 0.01 4. Medial digital extensor 61.36 1.44 15.75 0.05 30.14 1.30 09.45 0.02 5. Common digital extensor 80.44 1.66 40.00 0.10 49.20 1.45 20.00 0.02 6. Lateral digital extensor 40.15 1.45 20.00 0.07 2950 1.30 10.75 0.01 B. Caudo-medial group: 1. Gastrocnemius 50.00 1.55 07.88 0.05 29.96 1.30 16.35 0.03 2. Achilles tendon 47.25 1.40 05.76 0.02 39.42 .1.35 09.75 0.03 3. Superficial digital flexor 55.26 1.35 07.58 0.02 24.50 1.20 09.05 0.02 4. Deep digital flexor 50.79 1.20 19.25 0.03 22.81 1.30 13.00 0.02 -------------------------------------------------------------------------------------------------------------------The mean values of the measurements differed significantly between the age (P<0.01) and the limb groups (P<0.05)

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Table 3: Young` s modulus of elasticity ( E, kN / sq mm ), Factor of safety ( FS) and Fracture toughness ( FT, kN / mm 3/2 ) of tendons of the buffalo. __________________________________________________________________________ Name of muscle (tendon) Buffalo calf Adult buffalo ( No. of animals = 6) E FS FT E FS FT I. Forelimb: A. Cranio-lateral group: 1. Extensor carpii radialis 10.42 0.25 6 180 5 20.83 0.24 4 258 10 2. Medial digital extensor 9.23 0.20 3 170 2 20.20 0.25 2 250 10 3. Common digital extensor 9.50 0.10 2 170 2 18.00 0.10 2 240 15 4. Lateral digital extensor 10.17 0.20 9 180 5 20.15 0.25 2 254 15 B. Caudo-medial group: 1. Flexor carpii radialis 8.80 0.35 2 165 3 18.10 0.30 4 235 10 2. Flexor carpii ulnaris 8.10 0/20 2 160 5 19.00 0.20 3 240 10 3. Ulnaris lateralis 9.35 0.25 2 185 3 18.25 0.20 2 250 17 4. superficial digital flexor 10.88 0.15 5 180 4 20.80 0.20 3 240 12 5. Deep digital flexor 10.40 0.15 6 180 4 20.80 0.20 3 250 10 II. Hind limb A. Cranio-lateral group: 1. Peroneus tertius 7.90 0.25 12 160 5 18.35 0.30 3 303 15 2. Peroneus longus 8.25 0.10 4 160 4 18.15 10 3 215 10 3. Tibialis cranialis 8.05 0.15 7 160 5 17.00 0.30 3 370 15 4. Medial digital extensor 9.97 0.35 2 180 10 20.50 0.30 3 500 15 5. Common digital extensor 9.10 0.20 2 170 8 19.80 0.30 3 450 10 6. Lateral digital extensor 8.90 0.25 3 165 3 19.40 0.30 3 249 10 B. Caudo-medial group: 1. Gastrocnemius 10.40 0.40 7 180 5 20.35 0.20 2 440 15 2. Achilles tendon 12.75 0.30 9 200 10 21.35 0.15 4 315 10 3. Superficial digital flexor 10.20 0.25 8 180 5 20.00 0.25 3 570 15 4. Deep digital flexor 10.10 0.25 3 180 5 20.75 0.30 2 570 15 ------------------------------------------------------------------------------------------------------------------The mean values of the measurements differed significantly between the age (P<0.01) and the limb groups (P<0.05).

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Table 4: Critical length of micro-damage ( CL, mm ) and number of cycles causing fatigue failire(10 3 C 1 N/mm3/2) of the tendons of the buffalo. ________________________________________________________________________ Name of muscle (tendon) Buffalo Calf Adult buffalo ( No. of animals = 6) CL N CL N I. Forelimb: A. Cranio-lateral group: 1. Extensor carpii radialis 6.50 0.05 120 10 22.10 1.5 270 16 2. Medial digital extensor 10.0 0.05 10 02 16.30 1.2 40 06 3. Common digital extensor 0.86 0.01 10 01 12.05 1.1 40 03 4. Lateral digital extensor 1.45 0.01 10 01 36.45 1.4 160 10 B. Caudo-medial group: 1. Flexor carpii radialis 7.02 0.02 30 01 29.68 1.2 540 15 2. Flexor carpii ulnaris 3.00 0.01 10 01 27.00 1.1 350 13 3. Ulnaris lateralis 2.00 0.01 02 0.1 20.16 1.1 90 15 4. Superficial digital flexor 3.84 0.02 30 02 29.00 1.1 350 18 5. Deep digital flexor 4.05 0.01 40 03 32.58 1.2 400 20 II. Hind limb A. Cranio-lateral group: 1. Peroneus tertius 4.68 0.01 300 16 15.29 1.9 160 10 2. Peroneus longus 5.85 0.02 60 05 11.56 0.9 50 05 3. Tibialis cranialis 4.79 0.01 160 12 30.01 1.3 300 12 4. Medial digital extensor 2.45 0.01 10 01 20.58 1.1 230 16 5. Common digital extensor 1.58 0.01 1 0.01 10.64 0.8 30 05 6. Lateral digital extensor 5.98 0.02 10 0.1 22.35 1.1 200 12 B. Caudo-medial group: 1. Gastrocnemius 4.00 0.02 60 07 24.05 1.1 30 02 2. Achilles tendon 6.00 0.02 180 12 14.78 0.6 40 03 3. Superficial digital flexor 3.05 0.01 50 02 33.85 1.3 10 01 4. Deep digital flexor 4.15 0.01 10 01 40.10 1.7 10 01 -------------------------------------------------- --------------------------------------------------------------The mean values of the measurements differed significantly between the age (P<0.01) and the limb groups (P<0.05).