Assessing the role of surface layer and molecular probe size in diffusion within meniscus tissue.

Diffusion within extracellular matrix is essential to deliver nutrients and larger metabolites to the avascular region of the meniscus. It is well known that both structure and composition of the meniscus vary across its regions; therefore, it is crucial to fully understand how the heterogenous meniscal architecture affects its diffusive properties. The objective of this study was to investigate the effect of meniscal region (core tissue, femoral, and tibial surface layers) and molecular weight on the diffusivity of several molecules in porcine meniscus. Tissue samples were harvested from the central area of porcine lateral menisci. Diffusivity of fluorescein (MW 332 Da) and three fluorescence-labeled dextrans (MW 3k, 40k, and 150k Da) was measured via fluorescence recovery after photobleaching. Diffusivity was affected by molecular size, decreasing as the Stokes' radius of the solute increased. There was no significant effect of meniscal region on diffusivity for fluorescein, 3k and 40k dextrans (p>0.05). However, region did significantly affect the diffusivity of 150k Dextran, with that in the tibial surface layer being larger than in the core region (p = 0.001). Our findings contribute novel knowledge concerning the transport properties of the meniscus fibrocartilage. This data can be used to advance the understanding of tissue pathophysiology and explore effective approaches for tissue restoration.

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.