Skip to main content
Top
Published in: BMC Musculoskeletal Disorders 1/2012

Open Access 01-12-2012 | Research article

Stochastic amplitude-modulated stretching of rabbit flexor digitorum profundus tendons reduces stiffness compared to cyclic loading but does not affect tenocyte metabolism

Authors: Thomas H Steiner, Alexander Bürki, Stephen J Ferguson, Benjamin Gantenbein-Ritter

Published in: BMC Musculoskeletal Disorders | Issue 1/2012

Login to get access

Abstract

Background

It has been demonstrated that frequency modulation of loading influences cellular response and metabolism in 3D tissues such as cartilage, bone and intervertebral disc. However, the mechano-sensitivity of cells in linear tissues such as tendons or ligaments might be more sensitive to changes in strain amplitude than frequency. Here, we hypothesized that tenocytes in situ are mechano-responsive to random amplitude modulation of strain.

Methods

We compared stochastic amplitude-modulated versus sinusoidal cyclic stretching. Rabbit tendon were kept in tissue-culture medium for twelve days and were loaded for 1h/day for six of the total twelve culture days. The tendons were randomly subjected to one of three different loading regimes: i) stochastic (2 – 7% random strain amplitudes), ii) cyclic_RMS (2–4.42% strain) and iii) cyclic_high (2 - 7% strain), all at 1 Hz and for 3,600 cycles, and one unloaded control.

Results

At the end of the culture period, the stiffness of the “stochastic” group was significantly lower than that of the cyclic_RMS and cyclic_high groups (both, p < 0.0001). Gene expression of eleven anabolic, catabolic and inflammatory genes revealed no significant differences between the loading groups.

Conclusions

We conclude that, despite an equivalent metabolic response, stochastically stretched tendons suffer most likely from increased mechanical microdamage, relative to cyclically loaded ones, which is relevant for tendon regeneration therapies in clinical practice.
Appendix
Available only for authorised users
Literature
1.
go back to reference Wilson JJ, Best TM: Common overuse tendon problems: A review and recommendations for treatment. Am Fam Physician. 2005, 72 (5): 811-818.PubMed Wilson JJ, Best TM: Common overuse tendon problems: A review and recommendations for treatment. Am Fam Physician. 2005, 72 (5): 811-818.PubMed
2.
go back to reference Asundi KR, Kursa K, Lotz J, Rempel DM: In vitro system for applying cyclic loads to connective tissues under displacement or force control. Ann Biomed Eng. 2007, 35 (7): 1188-1195. 10.1007/s10439-007-9295-9.CrossRefPubMed Asundi KR, Kursa K, Lotz J, Rempel DM: In vitro system for applying cyclic loads to connective tissues under displacement or force control. Ann Biomed Eng. 2007, 35 (7): 1188-1195. 10.1007/s10439-007-9295-9.CrossRefPubMed
3.
go back to reference Sharma P, Maffulli N: Biology of tendon injury: healing, modeling and remodeling. J Musculoskelet Neuronal Interact. 2006, 6 (2): 181-190.PubMed Sharma P, Maffulli N: Biology of tendon injury: healing, modeling and remodeling. J Musculoskelet Neuronal Interact. 2006, 6 (2): 181-190.PubMed
4.
go back to reference Wang JH: Mechanobiology of tendon. J Biomech. 2006, 39 (9): 1563-1582. 10.1016/j.jbiomech.2005.05.011.CrossRefPubMed Wang JH: Mechanobiology of tendon. J Biomech. 2006, 39 (9): 1563-1582. 10.1016/j.jbiomech.2005.05.011.CrossRefPubMed
5.
go back to reference Nakama LH, King KB, Abrahamsson S, Rempel DM: Evidence of tendon microtears due to cyclical loading in an in vivo tendinopathy model. J Orthop Res. 2005, 23 (5): 1199-1205. 10.1016/j.orthres.2005.03.006.CrossRefPubMed Nakama LH, King KB, Abrahamsson S, Rempel DM: Evidence of tendon microtears due to cyclical loading in an in vivo tendinopathy model. J Orthop Res. 2005, 23 (5): 1199-1205. 10.1016/j.orthres.2005.03.006.CrossRefPubMed
6.
go back to reference Lavagnino M, Arnoczky SP, Tian T, Vaupel Z: Effect of amplitude and frequency of cyclic tensile strain on the inhibition of MMP-1 mRNA expression in tendon cells: an in vitro study. Connect Tissue Res. 2003, 44 (3–4): 181-187.CrossRefPubMed Lavagnino M, Arnoczky SP, Tian T, Vaupel Z: Effect of amplitude and frequency of cyclic tensile strain on the inhibition of MMP-1 mRNA expression in tendon cells: an in vitro study. Connect Tissue Res. 2003, 44 (3–4): 181-187.CrossRefPubMed
7.
go back to reference Lavagnino M, Arnoczky SP, Kepich E, Caballero O, Haut RC: A finite element model predicts the mechanotransduction response of tendon cells to cyclic tensile loading. Biomech Model Mechanobiol. 2008, 7 (5): 405-416. 10.1007/s10237-007-0104-z.CrossRefPubMed Lavagnino M, Arnoczky SP, Kepich E, Caballero O, Haut RC: A finite element model predicts the mechanotransduction response of tendon cells to cyclic tensile loading. Biomech Model Mechanobiol. 2008, 7 (5): 405-416. 10.1007/s10237-007-0104-z.CrossRefPubMed
8.
go back to reference Gardner K, Arnoczky SP, Lavagnino M: Effect of in vitro stress-deprivation and cyclic loading on the length of tendon cell cilia in situ. J Orthop Res. 2011, 29 (4): 582-587. 10.1002/jor.21271.CrossRefPubMed Gardner K, Arnoczky SP, Lavagnino M: Effect of in vitro stress-deprivation and cyclic loading on the length of tendon cell cilia in situ. J Orthop Res. 2011, 29 (4): 582-587. 10.1002/jor.21271.CrossRefPubMed
9.
go back to reference Hannafin JA, Arnoczky SP, Hoonjan A, Torzilli PA: Effect of stress deprivation and cyclic tensile loading on the material and morphologic properties of canine flexor digitorum profundus tendon: an in vitro study. J Orthop Res. 1995, 13 (6): 907-914. 10.1002/jor.1100130615.CrossRefPubMed Hannafin JA, Arnoczky SP, Hoonjan A, Torzilli PA: Effect of stress deprivation and cyclic tensile loading on the material and morphologic properties of canine flexor digitorum profundus tendon: an in vitro study. J Orthop Res. 1995, 13 (6): 907-914. 10.1002/jor.1100130615.CrossRefPubMed
10.
go back to reference Thornton GM, Shao X, Chung M, Sciore P, Boorman RS, Hart DA, Lo IK: Changes in mechanical loading lead to tendonspecific alterations in MMP and TIMP expression: influence of stress deprivation and intermittent cyclic hydrostatic compression on rat supraspinatus and Achilles tendons. Br J Sports Med. 2010, 44 (10): 698-703. 10.1136/bjsm.2008.050575.CrossRefPubMed Thornton GM, Shao X, Chung M, Sciore P, Boorman RS, Hart DA, Lo IK: Changes in mechanical loading lead to tendonspecific alterations in MMP and TIMP expression: influence of stress deprivation and intermittent cyclic hydrostatic compression on rat supraspinatus and Achilles tendons. Br J Sports Med. 2010, 44 (10): 698-703. 10.1136/bjsm.2008.050575.CrossRefPubMed
11.
go back to reference Arnoczky SP, Tian T, Lavagnino M, Gardner K: Ex vivo static tensile loading inhibits MMP-1 expression in rat tail tendon cells through a cytoskeletally based mechanotransduction mechanism. J Orthop Res. 2004, 22 (2): 328-333. 10.1016/S0736-0266(03)00185-2.CrossRefPubMed Arnoczky SP, Tian T, Lavagnino M, Gardner K: Ex vivo static tensile loading inhibits MMP-1 expression in rat tail tendon cells through a cytoskeletally based mechanotransduction mechanism. J Orthop Res. 2004, 22 (2): 328-333. 10.1016/S0736-0266(03)00185-2.CrossRefPubMed
12.
go back to reference Qi J, Chi L, Bynum D, Banes AJ: Gap junctions in IL-1β-mediated cell survival response to strain. J Appl Physiol. 2011, 110 (5): 1425-1431. 10.1152/japplphysiol.00477.2010.CrossRefPubMed Qi J, Chi L, Bynum D, Banes AJ: Gap junctions in IL-1β-mediated cell survival response to strain. J Appl Physiol. 2011, 110 (5): 1425-1431. 10.1152/japplphysiol.00477.2010.CrossRefPubMed
13.
go back to reference Chandrashekar N, Slauterbeck J, Hashemi J: Effects of cyclic loading on the tensile properties of human patellar tendon. Knee. 2012, 19 (1): 65-68. 10.1016/j.knee.2010.11.014.CrossRefPubMed Chandrashekar N, Slauterbeck J, Hashemi J: Effects of cyclic loading on the tensile properties of human patellar tendon. Knee. 2012, 19 (1): 65-68. 10.1016/j.knee.2010.11.014.CrossRefPubMed
14.
go back to reference Malaviya P, Butler DL, Korvick DL, Proch FS: In vivo tendon forces correlate with activity level and remain bounded: evidence in a rabbit flexor tendon model. J Biomech. 1998, 31 (11): 1043-1049. 10.1016/S0021-9290(98)00123-7.CrossRefPubMed Malaviya P, Butler DL, Korvick DL, Proch FS: In vivo tendon forces correlate with activity level and remain bounded: evidence in a rabbit flexor tendon model. J Biomech. 1998, 31 (11): 1043-1049. 10.1016/S0021-9290(98)00123-7.CrossRefPubMed
15.
go back to reference Saber S, Zhang AY, Ki SH, Lindsey DP, Smith RL, Riboh J, Pham H, Chang J: Flexor tendon tissue engineering: bioreactor cyclic strain increases construct strength. Tissue Eng Part A. 2010, 16 (6): 2085-2090. 10.1089/ten.tea.2010.0032.CrossRefPubMed Saber S, Zhang AY, Ki SH, Lindsey DP, Smith RL, Riboh J, Pham H, Chang J: Flexor tendon tissue engineering: bioreactor cyclic strain increases construct strength. Tissue Eng Part A. 2010, 16 (6): 2085-2090. 10.1089/ten.tea.2010.0032.CrossRefPubMed
16.
go back to reference Wren TA, Beaupré GS, Carter DR: A model for loading-dependent growth, development, and adaptation of tendons and ligaments. J Biomech. 1998, 31 (2): 107-114.CrossRefPubMed Wren TA, Beaupré GS, Carter DR: A model for loading-dependent growth, development, and adaptation of tendons and ligaments. J Biomech. 1998, 31 (2): 107-114.CrossRefPubMed
17.
go back to reference Reno C, Marchuk L, Sciore P, Frank CB, Hart DA: Rapid isolation of total RNA from small samples of hypocellular, dense connective tissues. Biotechniques. 1997, 22 (6): 1082-1086.PubMed Reno C, Marchuk L, Sciore P, Frank CB, Hart DA: Rapid isolation of total RNA from small samples of hypocellular, dense connective tissues. Biotechniques. 1997, 22 (6): 1082-1086.PubMed
18.
go back to reference Dudli S, Haschtmann D, Ferguson SJ: Fracture of the vertebral endplates, but not equienergetic impact load, promotes disc degeneration in vitro. J Orthop Res. 2011, 30 (5): 809-816.CrossRefPubMed Dudli S, Haschtmann D, Ferguson SJ: Fracture of the vertebral endplates, but not equienergetic impact load, promotes disc degeneration in vitro. J Orthop Res. 2011, 30 (5): 809-816.CrossRefPubMed
19.
go back to reference Livak KJ, Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) Method. Methods. 2001, 25 (4): 402-408. 10.1006/meth.2001.1262.CrossRefPubMed Livak KJ, Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) Method. Methods. 2001, 25 (4): 402-408. 10.1006/meth.2001.1262.CrossRefPubMed
20.
go back to reference Enobakhare BO, Bader DL, Lee DA: Quantification of sulfated glycosaminoglycans in chondrocyte/alginate cultures, by use of 1,9-dimethylmethylene blue. Anal Biochem. 1996, 243 (1): 189-191. 10.1006/abio.1996.0502.CrossRefPubMed Enobakhare BO, Bader DL, Lee DA: Quantification of sulfated glycosaminoglycans in chondrocyte/alginate cultures, by use of 1,9-dimethylmethylene blue. Anal Biochem. 1996, 243 (1): 189-191. 10.1006/abio.1996.0502.CrossRefPubMed
21.
go back to reference Farndale RW, Buttle DJ, Barrett AJ: Improved quantitation and discrimination of sulphated glycosaminoglycans by use of dimethylmethylene blue 1. Biochim Biophys Acta. 1986, 883 (2): 173-177. 10.1016/0304-4165(86)90306-5.CrossRefPubMed Farndale RW, Buttle DJ, Barrett AJ: Improved quantitation and discrimination of sulphated glycosaminoglycans by use of dimethylmethylene blue 1. Biochim Biophys Acta. 1986, 883 (2): 173-177. 10.1016/0304-4165(86)90306-5.CrossRefPubMed
22.
go back to reference Parkinson J, Samiric T, Ilic MZ, Feller JCAA, Handley CJ: Change in proteoglycan metabolism is a characteristic of human patellar tendinopathy. Arthritis Rheum. 2010, 62 (10): 3028-3035. 10.1002/art.27587.CrossRefPubMed Parkinson J, Samiric T, Ilic MZ, Feller JCAA, Handley CJ: Change in proteoglycan metabolism is a characteristic of human patellar tendinopathy. Arthritis Rheum. 2010, 62 (10): 3028-3035. 10.1002/art.27587.CrossRefPubMed
23.
go back to reference Shim JW, Elder SH: Influence of cyclic hydrostatic pressure on fibrocartilaginous metaplasia of achilles tendon fibroblasts. Biomech Model Mechanobiol. 2006, 5 (4): 247-252. 10.1007/s10237-005-0013-y.CrossRefPubMed Shim JW, Elder SH: Influence of cyclic hydrostatic pressure on fibrocartilaginous metaplasia of achilles tendon fibroblasts. Biomech Model Mechanobiol. 2006, 5 (4): 247-252. 10.1007/s10237-005-0013-y.CrossRefPubMed
24.
go back to reference Leigh DR, Abreu EL, Derwin KA: Changes in gene expression of individual matrix metalloproteinases differ in response to mechanical unloading of tendon fascicles in explant culture. J Orthop Res. 2008, 26 (10): 1306-1312. 10.1002/jor.20650.CrossRefPubMed Leigh DR, Abreu EL, Derwin KA: Changes in gene expression of individual matrix metalloproteinases differ in response to mechanical unloading of tendon fascicles in explant culture. J Orthop Res. 2008, 26 (10): 1306-1312. 10.1002/jor.20650.CrossRefPubMed
25.
go back to reference Woon CY, Kraus A, Raghavan SS, Pridgen BC, Megerle K, Pham H, Chang J: Three-dimensional-construct bioreactor conditioning in human tendon tissue engineering. Tissue Eng Part A. 2011, 17 (19–20): 2561-2572.CrossRefPubMed Woon CY, Kraus A, Raghavan SS, Pridgen BC, Megerle K, Pham H, Chang J: Three-dimensional-construct bioreactor conditioning in human tendon tissue engineering. Tissue Eng Part A. 2011, 17 (19–20): 2561-2572.CrossRefPubMed
26.
go back to reference White KL, Camire LM, Parks BG, Corey WS, Hinton RY: Krackow locking stitch versus locking premanufactured loop stitch for soft-tissue fixation: a biomechanical study. Arthroscopy. 2010, 26 (12): 1662-1666. 10.1016/j.arthro.2010.05.013.CrossRefPubMed White KL, Camire LM, Parks BG, Corey WS, Hinton RY: Krackow locking stitch versus locking premanufactured loop stitch for soft-tissue fixation: a biomechanical study. Arthroscopy. 2010, 26 (12): 1662-1666. 10.1016/j.arthro.2010.05.013.CrossRefPubMed
27.
go back to reference Nordin M, Frankel VH: Basic biomechanics of the musculoskelettal system. Edited by: Butler J. 2001, Maryland US: Lippincott Williams & Wilkins Nordin M, Frankel VH: Basic biomechanics of the musculoskelettal system. Edited by: Butler J. 2001, Maryland US: Lippincott Williams & Wilkins
28.
go back to reference Alberts B, Johnson A, Lewis J, Raff M, Roberts K, Walter P: Molecular biology of the cell. 2002, Fourth Edition: Garland Alberts B, Johnson A, Lewis J, Raff M, Roberts K, Walter P: Molecular biology of the cell. 2002, Fourth Edition: Garland
29.
go back to reference Kjaer M: Role of extracellular matrix in adaptation of tendon and skeletal muscle to mechanical loading. Physiol Rev. 2004, 84 (2): 649-698. 10.1152/physrev.00031.2003.CrossRefPubMed Kjaer M: Role of extracellular matrix in adaptation of tendon and skeletal muscle to mechanical loading. Physiol Rev. 2004, 84 (2): 649-698. 10.1152/physrev.00031.2003.CrossRefPubMed
30.
go back to reference Scott A, Danielson P, Abraham T, Fong G, Sampaio AV, Underhill TM: Mechanical force modulates scleraxis expression in bioartificial tendons. J Musculoskelet Neuronal Interact. 2011, 11 (2): 124-132.PubMed Scott A, Danielson P, Abraham T, Fong G, Sampaio AV, Underhill TM: Mechanical force modulates scleraxis expression in bioartificial tendons. J Musculoskelet Neuronal Interact. 2011, 11 (2): 124-132.PubMed
31.
go back to reference Thomopoulos S, Das R, Birman V, Smith L, Ku K, Elson EL, Pryse KM, Marquez JP, Genin GM: Fibrocartilage tissue engineering: the role of the stress environment on cell morphology and matrix expression. Tissue Eng Part A. 2011, 17 (7–8): 1039-1053.CrossRefPubMedPubMedCentral Thomopoulos S, Das R, Birman V, Smith L, Ku K, Elson EL, Pryse KM, Marquez JP, Genin GM: Fibrocartilage tissue engineering: the role of the stress environment on cell morphology and matrix expression. Tissue Eng Part A. 2011, 17 (7–8): 1039-1053.CrossRefPubMedPubMedCentral
32.
go back to reference Maeda T, Sakabe T, Sunaga A, Sakai K, Rivera AL, Keene DR, Sasaki T, Stavnezer E, Iannotti J, Schweitzer R, Ilic D, Baskaran H, Sakai T: Conversion of mechanical force into TGF-β-mediated biochemical signals. Curr Biol. 2011, 21 (11): 933-941. 10.1016/j.cub.2011.04.007.CrossRefPubMedPubMedCentral Maeda T, Sakabe T, Sunaga A, Sakai K, Rivera AL, Keene DR, Sasaki T, Stavnezer E, Iannotti J, Schweitzer R, Ilic D, Baskaran H, Sakai T: Conversion of mechanical force into TGF-β-mediated biochemical signals. Curr Biol. 2011, 21 (11): 933-941. 10.1016/j.cub.2011.04.007.CrossRefPubMedPubMedCentral
33.
go back to reference Altman GH, Horan RL, Martin I, Farhadi J, Stark PR, Volloch V, Richmond JC, Vunjak-Novakovic G, Kaplan DL: Cell differentiation by mechanical stress. FASEB J. 2002, 16 (2): 270-272.PubMed Altman GH, Horan RL, Martin I, Farhadi J, Stark PR, Volloch V, Richmond JC, Vunjak-Novakovic G, Kaplan DL: Cell differentiation by mechanical stress. FASEB J. 2002, 16 (2): 270-272.PubMed
34.
go back to reference Sassoon AA, Ozasa Y, Chikenji T, Sun YL, Larson DR, Maas ML, Zhao C, Jen J, Amadio PC: Skeletal muscle and bone marrow derived stromal cells: A comparison of tenocyte differentiation capabilities. J Orthop Res. 2012, 30 (11): 1710-1718. 10.1002/jor.22135.CrossRefPubMedPubMedCentral Sassoon AA, Ozasa Y, Chikenji T, Sun YL, Larson DR, Maas ML, Zhao C, Jen J, Amadio PC: Skeletal muscle and bone marrow derived stromal cells: A comparison of tenocyte differentiation capabilities. J Orthop Res. 2012, 30 (11): 1710-1718. 10.1002/jor.22135.CrossRefPubMedPubMedCentral
35.
go back to reference Qi J, Dmochowski JM, Banes AN, Tsuzaki M, Bynum D, Patterson M, Creighton A, Gomez S, Tech K, Cederlund A, Banes AJ: Differential expression and cellular localization of novel isoforms of the tendon biomarker tenomodulin. J Appl Physiol. 2012, 113 (6): 861-871. 10.1152/japplphysiol.00198.2012.CrossRefPubMed Qi J, Dmochowski JM, Banes AN, Tsuzaki M, Bynum D, Patterson M, Creighton A, Gomez S, Tech K, Cederlund A, Banes AJ: Differential expression and cellular localization of novel isoforms of the tendon biomarker tenomodulin. J Appl Physiol. 2012, 113 (6): 861-871. 10.1152/japplphysiol.00198.2012.CrossRefPubMed
36.
go back to reference Asundi KR, Rempel DM: Cyclic loading inhibits expression of MMP-3 but not MMP-1 in an in vitro rabbit flexor tendon model. Clin Biomech (Bristol, Avon). 2008, 23 (1): 117-121. 10.1016/j.clinbiomech.2007.08.007.CrossRef Asundi KR, Rempel DM: Cyclic loading inhibits expression of MMP-3 but not MMP-1 in an in vitro rabbit flexor tendon model. Clin Biomech (Bristol, Avon). 2008, 23 (1): 117-121. 10.1016/j.clinbiomech.2007.08.007.CrossRef
Metadata
Title
Stochastic amplitude-modulated stretching of rabbit flexor digitorum profundus tendons reduces stiffness compared to cyclic loading but does not affect tenocyte metabolism
Authors
Thomas H Steiner
Alexander Bürki
Stephen J Ferguson
Benjamin Gantenbein-Ritter
Publication date
01-12-2012
Publisher
BioMed Central
Published in
BMC Musculoskeletal Disorders / Issue 1/2012
Electronic ISSN: 1471-2474
DOI
https://doi.org/10.1186/1471-2474-13-222

Other articles of this Issue 1/2012

BMC Musculoskeletal Disorders 1/2012 Go to the issue