Tendon healing in interleukin-4 and interleukin-6 knockout mice.

Cytokines have been shown to play an important role in tendon and ligament healing by regulating cellular differentiation and activity. The majority of studies that have investigated the role of cytokines in tendon and ligament healing have added them to injured tissue and assessed their effect. Because the efficacy of exogenously applying cytokines is dependent upon many factors such as the correct dosage, timing, and frequency, conflicting results are often reported. To avoid these factors, this study used transgenic mice with knockouts of interleukin-4 (IL4 -/-) and interleukin-6 (IL6 -/-) to investigate their role in tendon healing. Because of the reported roles of both of these cytokines in inflammation and fibroplasia, it was hypothesized that the order of organizational, geometric, and mechanical properties would be (greatest to least) injured IL6 -/-, injured control, and injured IL4 -/- mice. In addition, it was hypothesized that specific cytokines would be upregulated in each knockout group, but not compensate for the lack of IL-4 or IL-6. Mechanical and organizational properties of injured tendons from IL6 -/- mice were inferior to that of control and IL4 -/- mice despite the upregulation of the pro-inflammatory cytokine TNF-alpha. Temporal levels of IL-10 and IL-13 in the IL4 -/- mice resulted in comparable and even superior properties when compared to CTL mice. This study shows that IL-6 could not be compensated for and plays an important role in tendon healing. This study also supports the use of this animal model to further investigate tendon healing.

Tendon properties in interleukin-4 and interleukin-6 knockout mice.

Cytokines are known to play an important role in normal tendon development, function, and maintenance through interactions with fibroblasts and extracellular matrix proteins. However, the role of interleukins on normal tendon activity remains poorly understood. Previous studies that have researched the role of specific cytokines by exogenously applying them have often reported conflicting results. Therefore, a knockout mouse model was used to investigate the role of interleukins 4 and 6 on normal tendon organizational and biomechanical properties. It was hypothesized that interleukin-6 knockout (IL6 -/-) mice will display more organized collagen orientation and greater cross-sectional area and mechanical properties when compared to that of control mice. In addition, interleukin-4 knockout (IL4 -/-) mice will display the most disorganized collagen orientation and lowest cross-sectional area and mechanical properties. As hypothesized, IL6 -/- mice show a trend towards lower angular deviation (more organized) (p<0.1) when compared to IL4 -/- mice. In addition, the IL6 -/- mice show a trend towards a higher percent relaxation (p<0.1) and a significantly higher modulus (p<0.01) when compared to CTL and IL4 -/- mice. Unexpectedly, the IL6 -/- mice exhibited no significant differences in collagen fiber distribution and maximum stress from the other groups and actually had a smaller cross-sectional area than CTL mice (p<0.1). This study supports transgenic mice as an animal model for investigating how cytokines affect normal tendon properties. In addition, this study demonstrates that interleukins may play an important role in tendon development, function, and maintenance.

Investigating tendon fascicle structure-function relationships in a transgenic-age mouse model using multiple regression models.

Proper replacement or repair of damaged tendons or ligaments requires functionally engineered tissue that mimics their native mechanical properties. While tendon structure-function relationships are generally assumed, there exists little quantitative evidence of the roles of distinct tendon components in tendon function. Previous work has used linear correlations to assess the independent, univariate effects of one structural or one biochemical variable on mechanics. The current study's objective was to simultaneously and rigorously evaluate the relative contributions of seven different structural and compositional variables in predicting tissue mechanical properties through the use of multiple regression statistical models. Structural, biochemical, and mechanical analysis were all performed on tail tendon fascicles from different groups of transgenic mice, which provide a reproducible, noninvasive, in vivo model of changes in tendon structure and composition. Interestingly, glycosaminoglycan (GAG) content was observed to be the strongest predictor of mechanical properties. GAG content was also well correlated with collagen content and mean collagen fibril diameter. Collagen fibril area fraction was a significant predictor only of material properties. Therefore, in a large multivariate model, GAG content was the largest predictor of mechanical properties, perhaps both through direct influence and indirectly through its correlation with collagen content and fibril structure.

Strain-rate sensitive mechanical properties of tendon fascicles from mice with genetically engineered alterations in collagen and decorin.

Tendons have complex mechanical behaviors that are nonlinear and time dependent. It is widely held that these behaviors are provided by the tissue composition and structure. It is generally thought that type I collagen provides the primary elastic strength to tendon while proteoglycans, such as decorin, play a role in failure and viscoelastic properties. This study sought to quantify such structure-function relationships by comparing tendon mechanical properties between normal mice and mice genetically engineered for altered type I collagen content and absence of decorin. Uniaxial tensile ramp to failure experiments were performed on tail tendon fascicles at two strain rates, 0.5%/s and 50%/s. Mutations in type I collagen led to reduced failure load and stiffness with no changes in failure stress, modulus or strain rate sensitivity. Fascicles without decorin had similar elastic properties to normal fascicles, but reduced strain rate sensitivity. Fascicles from immature mice, with increased decorin content compared to adult fascicles, had inferior elastic properties but higher strain rate sensitivity. These results showed that tendon viscoelasticity is affected by decorin content but not by collagen alterations. This study provides quantitative evidence for structure-function relationships in tendon, including the role of proteoglycan in viscoelasticity.