Compression loading in vitro regulates proteoglycan synthesis by tendon fibrocartilage

doi:10.1016/0003-9861(92)90127-I

Archives of Biochemistry and Biophysics

Volume 298, Issue 1, October 1992, Pages 303-312

https://doi.org/10.1016/0003-9861(92)90127-I Get rights and content

Abstract

The regulation of proteoglycan synthesis in a fibrocartilaginous tissue by mechanical loading was assessed in vitro. Discs of bovine tendon fibrocartilage were loaded daily with unconfined, cyclic, uniaxial compression (5 s/min, 20 min/day) and the synthesis of large and small proteoglycans was measured by incorporation of [³⁵S]sulfate. All discs synthesized predominantly large proteoglycan when first placed in culture. After 2 weeks in culture nonloaded discs synthesized predominantly small proteoglycans whereas loaded discs continued to produce predominantly large proteoglycan. The turnover of ³⁵S-labeled proteoglycan was not significantly altered by the compression regime. Increased synthesis of large proteoglycans was induced by a 4-day compression regime following 21 days of culture without compression. Inclusion of cytochalasin B during compression mimicked this induction. Autoradiography demonstrated that cell proliferation was minimal and confined to the disc edges whereas ³⁵S-labeled proteoglycan synthesis occurred throughout the discs. These experiments demonstrate that mechanical compression can regulate synthesis of distinct proteoglycan types in fibrocartilage.

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The spatial arrangement and interactions of the extracellular matrix (ECM) components control the mechanical behavior of tissue at multiple length scales. Changes in microscale deformation mechanisms affect tissue function and are often hallmarks of remodeling and disease. Despite their importance, the deformation mechanisms that modulate the mechanical behavior of collagenous tissue, particularly in indentation and compression modes of deformation, remain poorly understood. Here, we develop an integrated computational and experimental approach to investigate the deformation mechanisms of collagenous tissue at the microscale. While the complex deformation arising from indentation with a spherical probe is often considered a pitfall rather than an opportunity, we leverage this orientation-dependent deformation to examine the shear-regulated interactions of collagen fibrils and the role of crosslinks in modulating these interactions. We specifically examine tendon and cervix, two tissues rich in collagen with quite different microstructures and mechanical functions. We find that interacting, crosslinked collagen fibrils resist microscale longitudinal compressive forces, while widely used constitutive models fail to capture this behavior. The reorientation of collagen fibrils tunes the compressive stiffness of complex tissues like cervix. This study offers new insights into the mechanical behavior of collagen fibrils during indentation, and more generally, under longitudinal compressive forces, and illustrates the mechanisms that contribute to the experimentally observed orientation-dependent mechanical behavior.
Remodeling and disease can affect the deformation and interaction of tissue constituents, and thus mechanical function of tissue. Yet, the microscale deformation mechanisms are not well characterized in many tissues. Here, we develop a combined experimental-computational approach to infer the microscale deformation mechanisms of collagenous tissues with very different functions: tendon and cervix. Results show that collagen fibrils resist microscale forces along their length, though widely-used constitutive models do not account for this mechanism. This deformation process partially modulates the compressive stiffness of complex tissues such as cervix. Computational modeling shows that crosslink-mediated shear deformations are central to this unexpected behavior. This study offers new insights into the deformation mechanisms of collagenous tissue and the function of collagen crosslinkers.
Characterisation of native and decellularised porcine tendon under tension and compression: A closer look at glycosaminoglycan contribution to tendon mechanics
2023, Journal of the Mechanical Behavior of Biomedical Materials
Decellularised porcine superflexor tendon (pSFT) has been characterised as a suitable scaffold for anterior cruciate ligament replacement, with dimensions similar to hamstring tendon autograft. However, decellularisation of tissues may reduce or damage extracellular matrix components, leading to undesirable biomechanical changes at a whole tissue scale. Although the role of collagen in tendons is well established, the mechanical contribution of glycosaminoglycans (GAGs) is less evident and could be altered by the decellularisation process. In this study, the contribution of GAGs to the tensile and compressive mechanical properties of pSFT was determined and whether decellularisation affected these properties by reducing GAG content or functionality.
PSFTs were either enzymatically treated using chondroitinase ABC to remove GAGs or decellularised using previously established methods. Native, GAG-depleted and decellularised pSFT groups were then subjected to quantitative assays and biomechanical characterisation. In tension, specimens underwent stress relaxation and strength testing. In compression, specimens underwent confined compression testing.
The GAG-depleted group was found to have circa 86% reduction of GAG content compared to native and decellularised groups. There was no significant difference in GAG content between native (3.75 ± 0.58 μg/mg) and decellularised (3.40 ± 0.37 μg/mg) groups. Stress relaxation testing discovered the time-independent and time-dependent relaxation moduli of the decellularised group were reduced ≥50% compared to native and GAG-depleted groups. However, viscoelastic behaviour of native and GAG-depleted groups resulted similar. Strength testing discovered no differences between native and GAG-depleted group's properties, albeit a reduction ∼20% for decellularised specimens' linear modulus and tensile strength compared to native tissue. In compression testing, the aggregate modulus was found to be circa 74% lower in the GAG-depleted group than the native and decellularised groups, while the zero-strain permeability was significantly higher in the GAG-depleted group (0.86 ± 0.65 mm⁴/N) than the decellularised group (0.03 ± 0.04 mm⁴/N).
The results indicate that GAGs may significantly contribute to the mechanical properties of pSFT in compression, but not in tension. Furthermore, the content and function of GAGs in pSFTs are unaffected by decellularisation and the mechanical properties of the tissue remain comparable to native tissue.
Ultrasound strain mapping of the mouse Achilles tendon during passive dorsiflexion
2022, Journal of Biomechanics
Citation Excerpt :
Specifically, impinged regions of tendon possess fibrocartilage-like structural and compositional features that are lost when impingement is removed and are absent in non-impinged areas (Malaviya et al., 2000; Gillard et al., 1979; Maffulli et al., 2006). In some cases, this tendon fibrocartilage may be a protective adaptation that helps tendons withstand compressive forces from bone (Docking et al., 2013; Wren et al., 2000; Vogel and Koob, 1989; Koob et al., 1992). However, excessive impingement has also been implicated as a causative factor in several tendinopathies, thus motivating interest in understanding the link between impingement and altered tendon phenotype (Cook and Purdam, 2012; Almekinders et al., 2003; Giori et al., 1993).
Immediately prior to inserting into bone, many healthy tendons experience impingement from nearby bony structures. However, super-physiological levels of impingement are implicated in insertional tendinopathies. Unfortunately, the mechanisms underlying the connection between impingement and tendon pathology remain poorly understood, in part due to the shortage of well-characterized animal models of impingement at clinically relevant sites. As a first step towards developing a model of excessive tendon impingement, the objective of this study was to characterize the mechanical strain environment in the mouse Achilles tendon insertion under passive dorsiflexion and confirm that – like humans – mice experience impingement of the tendon insertion from the calcaneus (heel bone) in dorsiflexed ankle positions. Based on previous work in humans, we hypothesized that during dorsiflexion, the mouse Achilles tendon insertion would experience high levels of transverse compressive strain due to calcaneal impingement. A custom-built loading platform was used to apply passive dorsiflexion, while an ultrasound transducer positioned over the Achilles tendon captured radiofrequency images. A non-rigid image registration algorithm was then used to map the transverse compressive strain based on the acquired ultrasound image sequences. Our results demonstrate that during passive dorsiflexion, transverse compressive strains were produced throughout the Achilles tendon, with significantly larger strain magnitudes at the tendon insertion than at the midsubstance. Furthermore, there was increasing transverse compressive strain observed within the Achilles tendon as a function of increasing dorsiflexion angle. This study enhances our understanding of the unique mechanical loading environment of the Achilles tendon under physiologically relevant conditions.
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2020, Clinical Biomechanics
Citation Excerpt :
Their coupling of high fixed charge density and charge-to-charge repulsion forces also provide resistance to high compressive loads (Berenson et al., 1996). Evidence that compressive loading of tendons regulates proteoglycan synthesis further emphasizes the importance of proteoglycans for tendon compressive integrity (Koob et al., 1992). The interaction of water molecules, free cations, and macromolecules with a fixed negative charge density such as collagen and proteoglycans play an essential role in tendon biomechanical properties.
Electromechanical reshaping is a novel, minimally invasive means to induce mechanical changes in connective tissues, and has the potential to be utilized in lieu of current orthopedic therapies that involve tendons and ligaments. Electromechanical reshaping delivers an electrical current to tissues while under mechanical deformation, causing in situ redox changes that produce reliably controlled and spatially limited mechanical and structural changes. In this study, we examine the feasibility of altering Young's modulus and inducing a shape deformation using an ex vivo bovine Achilles tendon model.
Tendon was mechanically deformed in two different modes: (1) elongation to assess for tensile modulus and (2) compression to assess for compressive modulus. Electromechanical reshaping was applied to tendon specimens via flat plate platinum electrodes (6 V, 3 min) while simultaneously under mechanical strain for 15 min.
In elongation mode, post-electromechanical reshaping samples demonstrated a significant decrease in Young's modulus compared to pretreatment samples (66.02 and 45.12 MPa, respectively, p < 0.0049). In compression mode, posttreatment samples illustrated a significant shape change, with an increase in diameter (10.62 to 11.36 mm, p < 0.05) and decrease in thickness (4.13 to 3.62 mm, p < 0.05).
Results demonstrated a tissue softening effect without lengthening deformation during elongation, and a shortening effect without compromising compressive stiffness during compression. Electromechanical reshaping's reliable, low-cost, and efficacious methodology in inducing mechanical and structural connective tissue modifications illustrates a potential for future alternative orthopedic applications. Future studies will optimize and refine electromechanical reshaping to address clinically relevant geometries and methods such as needle techniques.
Fibrous tissues growth and remodeling: Evolutionary micro-mechanical theory
2017, Journal of the Mechanics and Physics of Solids
Living fibrous tissues are composite materials having the unique ability to adapt their size, shape, structure and mechanical properties in response to external loading. This adaptation, termed growth and remodeling (G&R), occurs throughout life and is achieved via cell-induced turnover of tissue constituents where some are degraded and new ones are produced. Realistic mathematical modeling of G&R provides insight into the basic processes, allows for hypotheses testing, and constitutes an essential tool for establishing clinical thresholds of pathological remodeling and for the production of tissue substitutes aimed to achieve target structure and properties. In this study, a general 3D micro-mechanical multi-scale theory of G&R in fibrous tissue was developed which connects between the evolution of the tissue structure and properties, and the underlying mechano-biological turnover events of its constituents. This structural approach circumvents a fundamental obstacle in modeling growth mechanics since the growth motion is not bijective. The model was realized for a flat tissue under two biaxial external loadings using data-based parameter values. The predictions show close similarity to characteristics of remodeled adult tissue including its structure, anisotropic and non-linear mechanical properties, and the onset of in situ pre-strain and pre-stress. The results suggest that these important features of living fibrous tissues evolve as they grow.
Tissue-scale anisotropy and compressibility of tendon in semi-confined compression tests
2015, Journal of Biomechanics
Citation Excerpt :
This indicates that the compressive response of tendon tissue is physiologically relevant and suggests that PGs are potentially involved in the load bearing mechanisms. Only few compression experiments on tendons have been reported (Koob et al., 1992; Lee et al., 2000; Williams et al., 2008; Fang et al., 2014) in contrast to tensile testing, which has been applied on various length scales, reaching from single collagen molecules (Sun et al., 2002, 2004; Bozec and Horton, 2005) and fibrils (Graham et al., 2004; Shen et al., 2008; Svensson et al., 2010, 2012) over fascicles (Screen et al., 2004, 2013; Clemmer et al., 2010; Thorpe et al., 2012) to whole tendons and tendon segments (Wren et al., 2003; Lake et al., 2010; Rigozzi et al., 2009; Kahn et al., 2010, 2013). Many of the aforementioned studies were used to identify the material parameters of constitutive models establishing relations between tissue stress and strain.
In this study, porcine tendon tissue was tested with a dedicated semi-confined compression set-up that enables us to induce states of either fibrils in compression (mode I), tension (mode II) or at constant length (mode III), respectively. The results suggest that tendon tissue is compressible and demonstrates a significantly stiffer response in mode I than in mode III. This implies that the fibril direction remains the axis of transverse isotropy in compression and that it provides an anisotropic contribution to the tissue stress. These results, which are important for the development of constitutive models for tendon tissue, are discussed with respect to the hierarchical structure of the extracellular matrix.

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Compression loading in vitro regulates proteoglycan synthesis by tendon fibrocartilage

Abstract

Int. Rev. Cytol

J. Biomech

Cell

Exp. Cell Res

Matrix

Trans. Orthop. Res. Soc

Biochem. J

Trans. Orthop. Res. Soc