The complex inhomogeneous architecture of the human meniscal tissue at the micro and nano scale in the absence of artefacts introduced by sample treatments has not yet been fully revealed. The knowledge of the internal structure organization is essential to understand the mechanical functionality of the meniscus and its relationship with the tissue’s complex structure. In this work, we investigated human meniscal tissue structure using up-to-date non-invasive imaging techniques, based on multiphoton fluorescence and quantitative second harmonic generation microscopy complemented with Environmental Scanning Electron Microscopy measurements. Observations on 50 meniscal samples extracted from 6 human menisci (3 lateral and 3 medial) revealed fundamental features of structural morphology and allowed us to quantitatively describe the 3D organisation of elastin and collagen fibres bundles. 3D regular waves of collagen bundles are arranged in “honeycomb-like” cells that are comprised of pores surrounded by the collagen and elastin network at the micro-scale. This type of arrangement propagates from macro to the nanoscale.

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Polycaprolactone (PCL) is widely used in additive manufacturing for the construction of scaffolds for tissue engineering because of its good bioresorbability, biocompatibility, and processability. Nevertheless, its use is limited by its inadequate mechanical support, slow degradation rate and the lack of bioactivity and ability to induce cell adhesion and, thus, bone tissue regeneration. In this study, we fabricated 3D PCL scaffolds reinforced with a novel Mg-doped bioactive glass (Mg-BG) characterized by good mechanical properties and biological reactivity. An optimization of the printing parameters and scaffold fabrication was performed; furthermore, an extensive microtopography characterization by scanning electron microscopy and atomic force microscopy was carried out. Nano-indentation tests accounted for the mechanical properties of the scaffolds, whereas SBF tests and cytotoxicity tests using human bone-marrow-derived mesenchymal stem cells (BM-MSCs) were performed to evaluate the bioactivity and in vitro viability. Our results showed that a 50/50 wt% of the polymer-to-glass ratio provides scaffolds with a dense and homogeneous distribution of Mg-BG particles at the surface and roughness twice that of pure PCL scaffolds. Compared to pure PCL (hardness H = 35 ± 2 MPa and Young’s elastic modulus E = 0.80 ± 0.05 GPa), the 50/50 wt% formulation showed H = 52 ± 11 MPa and E = 2.0 ± 0.2 GPa, hence, it was close to those of trabecular bone. The high level of biocompatibility, bioactivity, and cell adhesion encourages the use of the composite PCL/Mg-BG scaffolds in promoting cell viability and supporting mechanical loading in the host trabecular bone.

In this study, we observe that the poromechanical parameters in human meniscus vary spatially throughout the tissue. The response is anisotropic and the porosity is functionally graded. To draw these conclusions, we measured the anisotropic permeability and the “aggregate modulus” of the tissue, i.e., the stiffness of the material at equilibrium, after the interstitial fluid has ceased flowing. We estimated those parameters within the central portion of the meniscus in three directions (i.e., vertical, radial and circumferential) by fitting an enhanced model on stress relation confined compression tests. We noticed that a classical biphasic model was not sufficient to reproduce the observed experimental behaviour. We propose a poroelastic model based on the assumption that the fluid flow inside the human meniscus is described by a fractional porous medium equation analogous to Darcy’s law, which involves fractional operators. The fluid flux is then time-dependent for a constant applied pressure gradient (in contrast with the classical Darcy’s law, which describes a time independent fluid flux relation). We show that a fractional poroelastic model is well-suited to describe the flow within the meniscus and to identify the associated parameters (i.e., the order of the time derivative and the permeability). The results indicate that mean values of λβ,β in the central body are λβ=5.5443×10−10m4Ns1−β, β=0.0434, while, in the posterior and anterior regions, are λβ=2.851×10−10m4Ns1−β, β=0.0326 and λβ=1.2636×10−10m4Ns1−β, β=0.0232, respectively. Furthermore, numerical simulations show that the fluid flux diffusion is facilitated in the central part of the meniscus and hindered in the posterior and anterior regions.

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