Stress-deprivation induces an up-regulation of versican and connexin-43 mRNA and protein synthesis and increased ADAMTS-1 production in tendon cells in situ.

Purpose: The proper function of the tenocyte network depends on cell-matrix as well as intercellular communication that is mechanosensitive. Building on the concept that the etiopathogenic stimulus for tendon degeneration is the catabolic response of tendon cells to mechanobiologic under-stimulation, we studied the pericellular matrix rich in versican and its predominant proteolytic enzyme ADAMTS-1, as well as Connexin-43 (Cx43), a major gap junction forming protein in tendons, in stress-deprived rat tail tendon fascicles (RTTfs).Materials and Methods: RTTfs were stress-deprived for up to 7 days under tissue culture conditions. RT-qPCR was used to measure mRNA expression of versican, ADAMTS-1, and Cx43. Protein synthesis was determined using Western blotting and immunohistochemistry.Results: Stress-deprivation (SD) caused a statistically significant up-regulation of versican, ADAMTS-1, and Cx43 mRNA expression that was persistent over the 7-day test period. Western blot analysis and immunohistochemical assessment of protein synthesis revealed a marked increase of the respective proteins with SD. Inhibition of proteolytic enzyme activity with ilomastat prevented the increased versican degradation and Cx43 synthesis in 3 days stress-deprived tendons when compared with non-treated, stress-deprived tendons.Conclusion: In the absence of mechanobiological signaling the immediate pericellular matrix is modulated as tendon cells up-regulate their production of ADAMTS-1, and versican with subsequent proteoglycan degradation potentially leading to cell signaling cues increasing Cx43 gap junctional protein. The results also provide further support for the hypothesis that the cellular changes associated with tendinopathy are a result of decreased mechanobiological signaling and a loss of homeostatic cytoskeletal tension.

Effect of collagen length distribution and timing for repair on the active TGF-ß concentration in tendon.

The composition of extracellular matrix (ECM) in tendon depends on the secretion profile of resident cells known as tenocytes. For tissues with a mechanical role like tendon, mechanical strain is known to play an important role in determining the secretion profile of resident cells. Previously we explored the idea of estimating average concentrations of ECM molecules as a function of tendon strain magnitude and number of loading cycles. Specifically, we developed a model of the mechanical fatigue damage of tendon collagen fibers and introduced elementary cell responses (ECRs) by which local cellular-level responses to the strain environment, combined with the fatigue damage model, were scaled up to predict tissue-level responses. Using this approach, we demonstrated that the proposed model is capable of estimating average concentrations of ECM molecules that qualitatively accord with experimental observations. In this study, we increase model realism by extending this approach to consider the implications of a non-uniform collagen fiber distribution, and the influence of time delay on repair of damaged collagen fibers. Using this approach, we focus the study on the average tenocyte secretion profile for active transforming growth factor beta (TGF-ß), and discover that increasing fiber length dispersion and/or increasing repair delay leads to increasing active TGF-ß concentrations, and reduced sensitivity of average concentration profile of TGF-ß to tendon strain.

Predicting tenocyte expression profiles and average molecular concentrations in Achilles tendon ECM from tissue strain and fiber damage.

In this study, we propose a method for quantitative prediction of changes in concentrations of a number of key signaling, structural and effector molecules within the extracellular matrix of tendon. To achieve this, we introduce the notion of elementary cell responses (ECRs). An ECR defines a normal reference secretion profile of a molecule by a tenocyte in response to the tenocyte's local strain. ECRs are then coupled with a model for mechanical damage of tendon collagen fibers at different straining conditions of tendon and then scaled up to the tendon tissue level for comparison with experimental observations. Specifically, our model predicts relative changes in ECM concentrations of transforming growth factor beta, interleukin 1 beta, collagen type I, glycosaminoglycan, matrix metalloproteinase 1 and a disintegrin and metalloproteinase with thrombospondin motifs 5, with respect to tendon straining conditions that are consistent with the observations in the literature. In good agreement with a number of in vivo and in vitro observations, the model provides a logical and parsimonious explanation for how excessive mechanical loading of tendon can lead to under-stimulation of tenocytes and a degenerative tissue profile, which may well have bearing on a better understanding of tendon homeostasis and the origin of some tendinopathies.

Crimp length decreases in lax tendons due to cytoskeletal tension, but is restored with tensional homeostasis.

Collagen crimp morphology is thought to contribute to the material behavior of tendons and may reflect the local mechanobiological environment of tendon cells. Following loss of collagen tension in tendons, tenocytes initiate a contraction response that shortens tendon length which, in turn, may alter crimp patterns. We hypothesized that changes in the crimp pattern of tendons are the result of cell-based contractions which are governed by relative tautness/laxity of the collagen matrix. To determine the relationship between crimp pattern and tensional homeostasis, rat tail tendon fascicles (RTTfs) were either allowed to freely contract or placed in clamps with 10% laxity for 7 days. The freely contracting RTTfs showed a significant decrease in percent crimp length on both day 5 (3.66%) and day 7 (7.70%). This decrease in crimp length significantly correlated with the decrease in freely contracting RTTf length. Clamped RTTfs demonstrated a significant decrease in percent crimp length on day 5 (1.7%), but no significant difference in percent crimp length on day 7 (0.57%). The results demonstrate that the tendon crimp pattern appears to be under cellular control and is a reflection of the local mechanobiological environment of the extracellular matrix. The ability of tenocytes to actively alter the crimp pattern of collagen fibers also suggests that tenocytes can influence the viscoelastic properties of tendon. Understanding the interactions between tenocytes and their extracellular matrix may lead to further insight into the role tendon cells play in maintaining tendon heath and homeostasis. © 2016 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 35:573-579, 2017.

Hypoxia inhibits primary cilia formation and reduces cell-mediated contraction in stress-deprived rat tail tendon fascicles.

BACKGROUND: Hypoxia, which is associated with chronic tendinopathy, has recently been shown to decrease the mechanosensitivity of some cells. Therefore, the purpose of this study was to determine the effect of hypoxia on the formation of elongated primary cilia (a mechanosensing organelle of tendon cells) in vitro and to determine the effect of hypoxia on cell-mediated contraction of stress-deprived rat tail tendon fascicles (RTTfs). METHODS: Tendon cells isolated from RTTfs were cultured under normoxic (21% O2) or hypoxic (1% O2) conditions for 24 hours. The cells were then stained for tubulin and the number of cells with elongated cilia counted. RTTfs from 1-month-old male Sprague-Dawley rats were also cultured under hypoxic and normoxic conditions for three days and tendon length measured daily. RESULTS: A significant (p=0.002) decrease in the percent of elongated cilia was found in cells maintained in hypoxic conditions (54.1%±12.2) when compared in normoxic conditions (71.7%±6.32). RTTfs in hypoxia showed a significant decrease in the amount of contraction compared to RTTfs in normoxia after two (p=0.007) and three (p=0.001) days. CONCLUSION: The decreased incidence of elongated primary cilia in a hypoxic environment, as well as the decreased mechanoresponsiveness of tendon cells under these conditions may relate to the inability of some cases of chronic tendinopathy to respond to strain-based rehabilitation modalities (i.e. eccentric loading).

Increased susceptibility of skin from HERDA (Hereditary Equine Regional Dermal Asthenia)-affected horses to bacterial collagenase degradation: a potential contributing factor to the clinical signs of HERDA.

BACKGROUND: Hereditary equine regional dermal asthenia (HERDA) is a genetic disorder of collagen resulting in fragile, hyper-extensible skin and ulcerative lesions. The predominance of skin lesions have been shown to occur on the dorsum of HERDA-affected horses. While this has been postulated to be due to increased exposure to sunlight of these areas, the precise pathological mechanism which causes this to occur is unclear. HYPOTHESIS/OBJECTIVES: We hypothesized that an increase in collagenase activity, that has been associated with the exposure of dermal fibroblasts to sunlight, will significantly degrade the material properties of skin from HERDA-affected horses when compared to unaffected controls. ANIMALS: Six unaffected and seven HERDA-affected horses, all euthanized for other reasons. METHODS: Full-thickness skin samples from similar locations on each horse were collected and cut into uniform strips and their material properties (tensile modulus) determined by mechanical testing before (n = 12 samples/horse) or after (n = 12 samples/horse) incubation in bacterial collagenase at 37°C for 6 h. The change in modulus following treatment was then compared between HERDA-affected and unaffected horses using a Student's t-test. RESULTS: The modulus of skin from HERDA-affected horses decreased significantly more than that from unaffected horses following collagenase treatment (54 ± 7% versus 30 ± 16%, P = 0.004). CONCLUSIONS AND CLINICAL IMPORTANCE: The significant decrease in the modulus of skin from HERDA-affected horses following collagenase exposure suggests that their altered collagen microarchitecture is more susceptible to enzymatic degradation and may explain the localization of skin lesions in HERDA-affected horses to those areas of the body most exposed to sunlight. These findings appear to support the previously reported benefits of sunlight restriction in HERDA-affected horses.

High magnitude, in vitro, biaxial, cyclic tensile strain induces actin depolymerization in tendon cells.

BACKGROUND: the cytoskeleton is a dynamic arrangement of actin filaments that maintain cell shape and are vital in mediating the mechanobiological response of the cell. METHODS: to determine the cytoskeletal response to varying in vitro, biaxial stretch amplitudes, rat-tail tendon cells were paired into control and cyclically strained groups of 4.75, 9.5, or 12% strain at 1 Hz for 2 hours and the actin cytoskeleton stained. The cells were analyzed for actin staining intensity as a measure of relative depolymerization and for cell shape. Collagenase gene expression was measured in cells undergoing 12% cyclic strain at 1 Hz for 24 hours. RESULTS: there was no significant difference in the degree of actin staining intensity between the control group and cells strained at either 4.75 or 9.5%. However, cells strained at 12% demonstrated a significant decrease in actin staining intensity (depolymerization) compared to control cells, increased collagenase expression by 81%, and a clear shift towards a more rounded cell shape. CONCLUSION: the results of this study demonstrate that the previously reported induction of collagenase activity associated with the application of high magnitude, in vitro, tensile strains may actually be a result of cytoskeletal depolymerization, which causes loss of tensional homeostasis and alteration of cell shape.

Thermal energy enhances cell-mediated contraction of lax rat tail tendon fascicles following exercise.

BACKGROUND: the application of thermal energy (TE) has shown promise in the treatment of tendinopathy. However, the precise mechanism(s) of action of this therapy is unclear. The loss of tendon cell homeostatic tension, due to loading-induced laxity, produces catabolic changes associated with tendinopathy. This catabolic activity can be inhibited through the re-establishment of a normal tensile environment via a cellular contraction mechanism. We hypothesized that application of TE will enhance the contraction rate of lax rat tail tendon fascicles (RTTfs) in an in vitro model. METHODS: following loading, 10 lax RTTfs from each mature rat (n=5) were treated once daily for 7 days with TE by replacing the culture media at 37°C (control) with 42°C media. Using calibrated photographs, RTTf lengths were measured daily. Additional RTTfs were utilized to investigate any changes in material (n=12) and/or histological (n=12) properties with TE. RESULTS: TE significantly increased the contraction rate of RTTfs (p>0.001) without altering the material or histological properties. CONCLUSION: these results demonstrate that TE significantly enhances the contraction rate of previously exercised tendons. The ability to more quickly re-establish a normal mechanobiological environment, thus minimizing any catabolic changes, may explain the beneficial effects reported with applied TE in tendinopathy treatment.

Tendon mechanobiology: Current knowledge and future research opportunities.

Tendons mainly function as load-bearing tissues in the muscloskeletal system; transmitting loads from muscle to bone. Tendons are dynamic structures that respond to the magnitude, direction, frequency, and duration of physiologic as well as pathologic mechanical loads via complex interactions between cellular pathways and the highly specialized extracellular matrix. This paper reviews the evolution and current knowledge of mechanobiology in tendon development, homeostasis, disease, and repair. In addition, we review several novel mechanotransduction pathways that have been identified recently in other tissues and cell types, providing potential research opportunities in the field of tendon mechanobiology. We also highlight current methods, models, and technologies being used in a wide variety of mechanobiology research that could be investigated in the context of their potential applicability for answering some of the fundamental unanswered questions in this field. The article concludes with a review of the major questions and future goals discussed during the recent ORS/ISMMS New Frontiers in Tendon Research Conference held on September 10 and 11, 2014 in New York City.

Tendon Contraction After Cyclic Elongation Is an Age-Dependent Phenomenon: In Vitro and In Vivo Comparisons.

BACKGROUND: Tendons are viscoelastic tissues that deform (elongate) in response to cyclic loading. However, the ability of a tendon to recover this elongation is unknown. HYPOTHESIS: Tendon length significantly increases after in vivo or in vitro cyclic loading, and the ability to return to its original length through a cell-mediated contraction mechanism is an age-dependent phenomenon. STUDY DESIGN: Controlled laboratory study. METHODS: In vitro, rat tail tendon fascicles (RTTfs) from Sprague-Dawley rats of 3 age groups (1, 3, and 12 months) underwent 2% cyclic strain at 0.17 Hz for 2 hours, and the percentages of elongation were determined. After loading, the RTTfs were suspended for 3 days under tissue culture conditions and photographed daily to determine the amount of length contraction. In vivo, healthy male participants (n = 29; age, 19-49 years) had lateral, single-legged weightbearing radiographs taken of the knee at 60° of flexion immediately before, immediately after, and 24 hours after completing eccentric quadriceps loading exercises on the dominant leg to fatigue. Measurements of patellar tendon length were taken from the radiographs, and the percentages of tendon elongation and subsequent contraction were calculated. RESULTS: In vitro, cyclic loading increased the length of all RTTfs, with specimens from younger (1 and 3 months) rats demonstrating significantly greater elongation than those from older (12 months) rats (P = .009). The RTTfs contracted to their original length significantly faster (P < .001) and in an age-dependent fashion, with younger animals contracting faster. In vivo, repetitive eccentric loading exercises significantly increased patellar tendon length (P < .001). Patellar tendon length decreased 24 hours after exercises (P < .001) but did not recover completely (P < .001). There was a weak but significant (R (2) = 0.203, P = .014) linear correlation between the amount of tendon contraction and age, with younger participants (<30 years) demonstrating significantly more contraction (P = .014) at 24 hours than older participants (>30 years). CONCLUSION: Cyclic tendon loading results in a significant increase in tendon elongation under both in vitro and in vivo conditions. Tendons in both conditions demonstrated an incomplete return to their original length after 24 hours, and the extent of this return was age dependent. CLINICAL RELEVANCE: The age- and time-dependent contraction of tendons, elongated after repetitive loading, could result in transient alterations in the mechanobiological environment of tendon cells. This, in turn, could induce the onset of catabolic changes associated with the pathogenesis of tendinopathy. These results suggest the importance of allowing time for contraction between bouts of repetitive exercise and may explain why age is a predisposing factor in tendinopathy.

Tendon cell ciliary length as a biomarker of in situ cytoskeletal tensional homeostasis.

To determine if tendon cell ciliary length could be used as a biomarker of cytoskeletal tensional homeostasis, 20 mm long adult rat tail tendons were placed in double artery clamps set 18 mm apart to create a 10% laxity. The tendons were allowed to contract over a 7 day period under culture conditions. At 0, 1, 5, and 7 days the tendon cell cilia were stained and ciliary length measured using confocal imaging. There was a significant (p<0.001) increase in ciliary length at 1 day. At day 5 (when the tendon became visibly taut) there was a significant (p<0.001) decrease in ciliary length compared to day 1. By day 7 the tendon remained taut and ciliary length returned to day zero values (p=0.883). These results suggest that cilia length reflects the local mechanobiological environment of tendon cells and could be used as a potential in situ biomarker of altered cytoskeletal tensional homeostasis.

Age-related changes in the cellular, mechanical, and contractile properties of rat tail tendons.

Tendon laxity following injury, cyclic creep, or repair has been shown to alter the normal homeostasis of tendon cells, which can lead to degenerative changes in the extracellular matrix. While tendon cells have been shown to have an inherent contractile mechanism that gives them some ability to retighten lax tendons and reestablish a homeostatic cellular environment, the effect of age on this process is unknown. To determine the effect of aging on cell number, cell shape, and tensile modulus on tendons as well as the rate of cell-mediated contraction of lax tendons, tail tendon fascicles from 1-, 3-, and 12-month-old rats were analyzed. Aging results in a decrease (p < 0.001) in cell number per mm(2): 1 m (981 ± 119), 3 m (570 ± 108), and 12 m (453 ± 23), a more flattened (p < 0.001) cell nuclei shape and a higher (p < 0.001) tensile modulus (MPa) of the tendons: 1 m (291 ± 2), 3 m (527 ± 38), and 12 m (640 ± 102). Both the extent and rate of contraction over 7 days decreased with age (p = 0.007). This decrease in contraction rate with age correlates to the observed changes seen in aging tendons [increased modulus (r(2) = 0.95), decreased cell number (r(2) = 0.89)]. The ability of tendons to regain normal tension following injury or exercise-induced laxity is a key factor in the recovery of tendon function. The decreased contraction rate as a function of age observed in the current study may limit the ability of tendon cells to retighten lax tendons in older individuals. This, in turn, may place these structures at further risk for injury or altered function.

Age-dependent effects of systemic administration of oxytetracycline on the viscoelastic properties of rat tail tendons as a mechanistic basis for pharmacological treatment of flexural limb deformities in foals.

OBJECTIVE: To describe the effect of systemically administered oxytetracycline on the viscoelastic properties of rat tail tendon fascicles (TTfs) to provide a mechanistic rationale for pharmacological treatment of flexural limb deformities in foals. SAMPLE: TTfs from ten 1-month-old and ten 6-month-old male Sprague-Dawley rats. PROCEDURES: 5 rats in each age group were administered oxytetracycline (50 mg/kg, IP, q 24 h) for 4 days. The remaining 5 rats in each age group served as untreated controls. Five days after initiation of oxytetracycline treatment, TTfs were collected and their viscoelastic properties were evaluated via a stress-relaxation protocol. Maximum modulus and equilibrium modulus were compared via a 2-way ANOVA. Collagen fibril size, density, and orientation in TTfs were compared between treated and control rats. RESULTS: Viscoelastic properties were significantly decreased in TTfs from 1-month-old oxytetracycline-treated rats, compared with those in TTfs from 1-month-old control rats. Oxytetracycline had no effect on the viscoelastic properties of TTfs from 6-month-old rats. Collagen fibril size, density, and orientation in TTfs from 1-month-old rats did not differ between oxytetracycline-treated and control rats. CONCLUSIONS AND CLINICAL RELEVANCE: Results confirmed that systemically administered oxytetracycline decreased the viscoelastic properties of TTfs from 1-month-old rats but not those of TTfs from 6-month-old rats. The decrease in viscoelastic properties associated with oxytetracycline treatment does not appear to be caused by altered collagen fibril diameter or organization. The age-dependent effect of oxytetracycline on the viscoelastic properties of tendons may be related to its effect on the maturation of the extracellular matrix of developing tendons.

Evaluation of intersegmental vertebral motion during performance of dynamic mobilization exercises in cervical lateral bending in horses.

OBJECTIVE: To identify differences in intersegmental bending angles in the cervical, thoracic, and lumbar portions of the vertebral column between the end positions during performance of 3 dynamic mobilization exercises in cervical lateral bending in horses. ANIMALS: 8 nonlame horses. PROCEDURES: Skin-fixed markers on the head, cervical transverse processes (C1-C6) and spinous processes (T6, T8, T10, T16, L2, L6, S2, and S4) were tracked with a motion analysis system with the horses standing in a neutral position and in 3 lateral bending positions to the left and right sides during chin-to-girth, chin-to-hip, and chin-to-tarsus mobilization exercises. Intersegmental angles for the end positions in the various exercises performed to the left and right sides were compared. RESULTS: The largest changes in intersegmental angles were at C6, especially for the chin-to-hip and chin-to-tarsus mobilization exercises. These exercises were also associated with greater lateral bending from T6 to S2, compared with the chin-to-girth mobilization or neutral standing position. The angle at C1 revealed considerable bending in the chin-to-girth position but not in the 2 more caudal positions. CONCLUSIONS AND CLINICAL RELEVANCE: The amount of bending in different parts of the cervical vertebral column differed among the dynamic mobilization exercises. As the horse's chin moved further caudally, bending in the caudal cervical and thoracolumbar regions increased, suggesting that the more caudal positions may be particularly effective for activating and strengthening the core musculature that is used to bend and stabilize the horse's back.

Re-establishment of cytoskeletal tensional homeostasis in lax tendons occurs through an actin-mediated cellular contraction of the extracellular matrix.

Cytoskeletal tensional homeostasis is known to be an important factor in controlling catabolic gene expression in tendon cells. Loss of cell tension in lax rat tail tendon fascicles (RTTfs) has been associated with an upregulation of MMP-13 gene expression and protein synthesis. To determine the role of the actin cytoskeleton in re-establishing tensional homeostasis in lax tendons, RTTfs were allowed to freely contract in vitro for 8 days. The cultured RTTfs contracted rapidly, reaching 50% of their initial length by 3 days. This contraction was associated with the presence of α-smooth muscle actin positive cells within the tendon. Disruption of the actin network by cytochalasian D caused an immediate and significant elongation of the contracted RTTfs. Subsequent removal of the cytochalasian D re-initiated the contraction process. When lax RTTfs were allowed to contract between fixed clamps in culture and become taut, they demonstrated a marked decrease in MMP-13 staining intensity when compared to freely contracting RTTfs. The ability of native tendon cells to contract lax tendons and re-establish their homeostatic "set point" with respect to collagenase production may be an important mechanism in the recovery of tendons elongated by injury, surgical positioning, or cyclic, viscoelastic creep secondary to repetitive exercise.

Resection of Grade III cranial horn tears of the equine medial meniscus alter the contact forces on medial tibial condyle at full extension: an in-vitro cadaveric study.

OBJECTIVE: To evaluate the magnitude and distribution of joint contact pressure on the medial tibial condyle after grade III cranial horn tears of the medial meniscus. STUDY DESIGN: Experimental study. ANIMALS: Cadaveric equine stifles (n = 6). METHODS: Cadaveric stifles were mounted in a materials testing system and electronic pressure sensors were placed between the medial tibial condyle and medial meniscus. Specimens were loaded parallel to the longitudinal axis of the tibia to 1800 N at 130°, 140°, 150°, and 160° stifle angle. Peak pressure and contact area were recorded from the contact maps. Testing was repeated after surgical creation of a grade III cranial horn tear of the medial meniscus, and after resection of the simulated tear. RESULTS: In the intact specimens, a significantly smaller contact area was observed at 160° compared with the other angles (P < .05). Creation of a grade III cranial horn tear in the medial meniscus did not significantly alter the pressure or contact area measurements at any stifle angle compared with intact specimens (P > .05). Resection of the tear resulted in significantly higher peak pressures in the central region of the medial tibial condyle at a stifle angle of 160° relative to the intact (P = .026) and torn (P = .012) specimens. CONCLUSIONS: Resection of grade III cranial horn tears in the medial meniscus resulted in a central focal region of increased pressure on the medial tibial condyle at 160° stifle angle.

Evaluation of biomechanical effects of four stimulation devices placed on the hind feet of trotting horses.

OBJECTIVE: To compare effects of 4 types of stimulation devices attached to the hind feet on hoof flight, joint angles, and net joint powers of trotting horses. ANIMALS: 8 clinically normal horses. PROCEDURES: Horses were evaluated under 5 conditions in random order: no stimulators, loose straps (10 g), lightweight tactile stimulators (55 g), limb weights (700 g), and limb weights with tactile stimulators (700 g). Reflective markers on the hind limbs were tracked during the swing phase of 6 trotting trials performed at consistent speed to determine peak hoof heights and flexion angles of the hip, stifle, tarsal, and metatarsophalangeal joints. Inverse dynamic analysis was used to calculate net joint energies. Comparisons among stimulators were made. RESULTS: Peak hoof height was lowest for no stimulators (mean ± SD, 5.42 ± 1.38 cm) and loose straps (6.72 ± 2.19 cm), intermediate for tactile stimulators (14.13 ± 7.33 cm) and limb weights (16.86 ± 15.93 cm), and highest for limb weights plus tactile stimulators (24.35 ± 13.06 cm). Compared with no stimulators, net tarsal energy generation increased for tactile stimulators, limb weights, and limb weights plus tactile stimulators, but only the weighted conditions increased net energy generation across the hip joint. CONCLUSIONS AND CLINICAL RELEVANCE: The type and weight of foot stimulators affected the magnitude of the kinematic and kinetic responses and the joints affected. These findings suggest that different types of foot stimulators are appropriate for rehabilitation of specific hind limb gait deficits, such as toe dragging and a short stride.

In situ deflection of tendon cell-cilia in response to tensile loading: an in vitro study.

To determine if a correlation exists between tensile loading and the deflection of tendon cell-cilia in situ, rat-tail tendon fascicles were stained for tubulin and mounted in a loading device attached to the stage of a confocal microscope. Individual tendon cells (n = 13) were identified and sequential images taken at 0%, 2%, 4%, 6%, and 8% grip to grip strain. The change in ciliary deflection angle was then measured at each strain level. To determine the ability of cilia to return to their original orientation, additional fascicles were loaded to 6% strain and then unloaded to 0% and tendon cell-ciliary (n = 10) deflection angle measured. There was a weak (r(2) = 0.40) but significant (p < 0.0001) correlation between the change in deflection angle and applied strain. Tensile loading produced a change in deflection angle from 0% to 3% (p = 0.039) and from 3% to 6% (p = 0.001) strain. There was no change (p = 1.000) in deflection angle from 6% to 8% strain. Reducing the strain from 6% to 0% resulted in a change (p = 0.048) in angle towards the pre-load position. However, the angle did not return to the pre-strain position (p = 0.025). These results demonstrate that tensile loading produces in situ deflection of tendon cell-cilia and supports the concept that cilia are involved in the mechanotransduction response of tendon cells.

Effect of in vitro stress-deprivation and cyclic loading on the length of tendon cell cilia in situ.

To determine the effect of loading conditions on the length of primary cilia in tendon cells in situ, freshly harvested rat tail tendons were stress-deprived (SD) for up to 72 h, cyclically loaded at 3% strain at 0.17 Hz for 24 h, or SD for 24 h followed by cyclic loading (CL) for 24 h. Tendon sections were stained for tubulin, and cilia measured microscopically. In fresh control tendons, cilia length ranged from 0.6 to 2.0 µm with a mean length of 1.1 µm. Following SD, cilia demonstrated an increase (p < 0.001) in overall length at 24 h when compared to controls. Cilia length did not increase with time of SD (p = 0.329). Cilia in cyclically loaded tendons were shorter (p < 0.001) compared to all SD time periods, but were not different from 0 time controls (p = 0.472). CL for 24 h decreased cilia length in 24 h SD tendons (p < 0.001) to levels similar to those of fresh controls (p = 0.274). The results of this study demonstrate that SD resulted in an immediate and significant increase in the length of primary cilia of tendon cells, which can be reversed by cyclic tensile loading. This suggests that, as in other tissues, cilia length in tendon cells is affected by mechanical signaling from the extracellular matrix.

Infrapatellar Straps Decrease Patellar Tendon Strain at the Site of the Jumper's Knee Lesion: A Computational Analysis Based on Radiographic Measurements.

BACKGROUND: The impetus for the use of patellar straps in the treatment of patellar tendinopathy has largely been based on empirical evidence and not on any mechanistic rationale. A computational model suggests that patellar tendinopathy may be a result of high localized tendon strains that occur at smaller patella-patellar tendon angles (PPTAs). HYPOTHESIS: Infrapatellar straps will decrease the mean localized computational strain in the area of the patellar tendon commonly involved in jumper's knee by increasing the PPTA. STUDY DESIGN: Controlled laboratory study. METHODS: Twenty adult males had lateral weightbearing and nonweightbearing radiographs of their knees taken with and without 1 of 2 infrapatellar straps at 60° of knee flexion. Morphologic measurements of PPTA and patellar tendon length with and without the straps were used as input data into a previously described computational model to calculate average and maximum strain at the common location of the jumper's knee lesion during a simulated jump landing. RESULTS: The infrapatellar bands decreased the predicted localized strain (average and maximum) in the majority of participants by increasing PPTA and/or decreasing patellar tendon length. When both PPTA and patellar tendon length were altered by the straps, there was a strong and significant correlation with the change in predicted average localized strain with both straps. CONCLUSION: Infrapatellar straps may limit excessive patella tendon strain at the site of the jumper's knee lesion by increasing PPTA and decreasing patellar tendon length rather than by correcting some inherent anatomic or functional abnormality in the extensor apparatus. CLINICAL RELEVANCE: The use of infrapatellar straps may help prevent excessive localized tendon strains at the site of the jumper's knee lesion during a jump landing.