Tensile energy dissipation and mechanical properties of the knee meniscus: relationship with fiber orientation, tissue layer, and water content.

Introduction: The knee meniscus distributes and dampens mechanical loads. It is composed of water (∼70%) and a porous fibrous matrix (∼30%) with a central core that is reinforced by circumferential collagen fibers enclosed by mesh-like superficial tibial and femoral layers. Daily loading activities produce mechanical tensile loads which are transferred through and dissipated by the meniscus. Therefore, the objective of this study was to measure how tensile mechanical properties and extent of energy dissipation vary by tension direction, meniscal layer, and water content. Methods: The central regions of porcine meniscal pairs (n = 8) were cut into tensile samples (4.7 mm length, 2.1 mm width, and 0.356 mm thickness) from core, femoral and tibial components. Core samples were prepared parallel (circumferential) and perpendicular (radial) to the fibers. Tensile testing consisted of frequency sweeps (0.01-1Hz) followed by quasi-static loading to failure. Dynamic testing yielded energy dissipation (ED), complex modulus (E*), and phase shift (δ) while quasi-static tests yielded Young's Modulus (E), ultimate tensile strength (UTS), and strain at UTS (εUTS). To investigate how ED is influenced by the specific mechanical parameters, linear regressions were performed. Correlations between sample water content (φw) and mechanical properties were investigated. A total of 64 samples were evaluated. Results: Dynamic tests showed that increasing loading frequency significantly reduced ED (p < 0.05). Circumferential samples had higher ED, E*, E, and UTS than radial ones (p < 0.001). Stiffness was highly correlated with ED (R2 > 0.75, p < 0.01). No differences were found between superficial and circumferential core layers. ED, E*, E, and UTS trended negatively with φw (p < 0.05). Discussion: Energy dissipation, stiffness, and strength are highly dependent on loading direction. A significant amount of energy dissipation may be associated with time-dependent reorganization of matrix fibers. This is the first study to analyze the tensile dynamic properties and energy dissipation of the meniscus surface layers. Results provide new insights on the mechanics and function of meniscal tissue.

Effect of molecular weight and tissue layer on solute partitioning in the knee meniscus.

Objective: Knee meniscus tissue is partly vascularized, meaning that nutrients must be transported through the extracellular matrix of the avascular portion to reach resident cells. Similarly, drugs used as therapeutic agents to treat meniscal pathologies rely on transport through the tissue. The driving force of diffusive transport is the gradient of concentration, which depends on molecular solubility. The meniscus is organized into a core region sandwiched between the tibial and femoral superficial layers. Structural differences exist across meniscal regions; therefore, regional differences in solubility are also hypothesized. Methods: Samples from the core, tibial and femoral layers were obtained from 5 medial and 5 lateral porcine menisci. The partition coefficient (K) of fluorescein, 3 â€‹kDa and 40 â€‹kDa dextrans in the layers of the meniscus was measured using an equilibration experiment. The effect of meniscal compartment, layer, and solute molecular weight on K was analyzed using a three-way ANOVA. Results: K ranged from a high of ∼2.9 in fluorescein to a low of ∼0.1 in 40 â€‹kDa dextran and was inversely related to the solute molecular weight across all tissue regions. Tissue layer only had a significant effect on partitioning of 40k Dex solute, which was lower in the tibial surface layer relative to the core (p â€‹= â€‹0.032). Conclusion: This study provides insight into depth-dependent partitioning in the meniscus, indicating the limiting effect of the meniscus superficial layer on solubility increases with solute molecular size. This illustrates how the surface layers could potentially reduce the effectiveness of drug delivery therapies incorporating large molecules (>40 â€‹kDa).

Heterogeneity of dynamic shear properties of the meniscus: A comparison between tissue core and surface layers.

Damage to the meniscus has been associated with excessive shear loads. Aimed at elucidating meniscus pathophysiology, previous studies have investigated the shear properties of the meniscus fibrocartilaginous core. However, the meniscus is structurally inhomogeneous, with an external cartilaginous envelope (tibial and femoral surface layers) wrapping the tissue core. To date, little is known about the shear behavior of the surface layers. The objective of this study was to measure the dynamic shear properties of the surface layers and derive empirical relations with their composition. Specimens were harvested from tibial and femoral surface layers and core of porcine menisci (medial and lateral, n = 10 each). Frequency sweep tests yielded complex shear modulus (G*) and phase shifts (δ). Mechanical behavior of regions was described by a generalized Maxwell model. Correlations between shear moduli with water and glycosaminoglycans content of the tissue regions were investigated. The femoral surface had the lowest shear modulus, when compared to core and tibial regions. A 3-relaxation times Maxwell model satisfactorily interpreted the shear behavior of all tissue regions. Inhomogeneous tissue composition was also observed, with water content in the surface layers being higher when compared with tissue core. Water content negatively correlated with shear properties in all regions. The lower measured shear properties in the femoral layer may explain the higher prevalence of meniscal tears on the superior surface of the tissue. The heterogenous behavior of the tissue in shear provides insight into meniscus pathology and has important implications for efforts to tissue engineer replacement tissues.

Strain-Dependent Diffusivity of Small and Large Molecules in Meniscus.

Due to lack of full vascularization, the meniscus relies on diffusion through the extracellular matrix to deliver small (e.g., nutrients) and large (e.g., proteins) to resident cells. Under normal physiological conditions, the meniscus undergoes up to 20% compressive strains. While previous studies characterized solute diffusivity in the uncompressed meniscus, to date, little is known about the diffusive transport under physiological strain levels. This information is crucial to fully understand the pathophysiology of the meniscus. The objective of this study was to investigate strain-dependent diffusive properties of the meniscus fibrocartilage. Tissue samples were harvested from the central portion of porcine medial menisci and tested via fluorescence recovery after photobleaching to measure diffusivity of fluorescein (332 Da) and 40 K Da dextran (D40K) under 0%, 10%, and 20% compressive strain. Specifically, average diffusion coefficient and anisotropic ratio, defined as the ratio of the diffusion coefficient in the direction of the tissue collagen fibers to that orthogonal, were determined. For all the experimental conditions investigated, fluorescein diffusivity was statistically faster than that of D40K. Also, for both molecules, diffusion coefficients significantly decreased, up to ∼45%, as the strain increased. In contrast, the anisotropic ratios of both molecules were similar and not affected by the strain applied to the tissue. This suggests that compressive strains used in this study did not alter the diffusive pathways in the meniscus. Our findings provide new knowledge on the transport properties of the meniscus fibrocartilage that can be leveraged to further understand tissue pathophysiology and approaches to tissue restoration.

Compressive Properties and Hydraulic Permeability of Human Meniscus: Relationships With Tissue Structure and Composition.

The meniscus is crucial in maintaining knee function and protecting the joint from secondary pathologies, including osteoarthritis. The meniscus has been shown to absorb up to 75% of the total load on the knee joint. Mechanical behavior of meniscal tissue in compression can be predicted by quantifying the mechanical parameters including; aggregate modulus (H) and Poisson modulus (ν), and the fluid transport parameter: hydraulic permeability (K). These parameters are crucial to develop a computational model of the tissue and for the design and development of tissue engineered scaffolds mimicking the native tissue. Hence, the objective of this study was to characterize the mechanical and fluid transport properties of human meniscus and relate them to the tissue composition. Specimens were prepared from the axial and the circumferential anatomical planes of the tissue. Stress relaxation tests yielded the H, while finite element modeling was used to curve fit for ν and K. Correlations of moduli with water and glycosaminoglycans (GAGs) content were investigated. On average H was found to be 0.11 ± 0.078 MPa, ν was 0.32 ± 0.057, and K was 2.9 ± 2.27 × 10-15 m4N-1s-1. The parameters H, ν, and K were not found to be statistically different across compression orientation or compression level. Water content of the tissue was 77 ± 3.3% while GAG content was 8.79 ± 1.1%. Interestingly, a weak negative correlation was found between H and water content (R2 ~ 34%) and a positive correlation between K and GAG content (R2 ~ 53%). In conclusion, while no significant differences in transport and compressive properties can be found across sample orientation and compression levels, data trends suggest potential relationships between magnitudes of H and K, and GAG content.