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The meniscus has complex biomechanical functions endowed by the hierarchical fibrous structure of its extracellular matrix (ECM), which plays a central role in protecting the knee joint. However, it is challenging to recapitulate the ECM structure of the meniscus. Herein, we used electrospinning to prepare various scaffolds with distinct nanofibrous structures to approximate that of the heterogeneous ultrastructure of meniscus ECM. Our results showed that adjusting the deposition manner of nanofibers during electrospinning could effectively manipulate the architectures of resulting scaffolds. This approach led to electrospun scaffolds with random or aligned fibrous structures that reassemble the surface or superficial layers of the meniscus. We also showed that assembly of electrospun nanofibers into yarn-like configurations replicates the circumferentially aligned fibrous bundles of the inner part of the meniscus. These scaffolds exhibited distinct fibrous anisotropies and mechanical properties. In vitro studies indicated good cytocompatibility of scaffolds, especially, the yarn scaffold supported considerable meniscus cell infiltration. The meniscus cells, in turn, enhanced the mechanical properties of scaffolds. Taken together, these data imply that electrospun scaffolds may have potential in enhancing meniscus repair and regeneration., (© 2020 Wiley Periodicals LLC.)

Background: Meniscus tissue engineering has provided a great potential treatment for meniscal injuries. However, few scaffolds in meniscus tissue engineering have matched the mechanical properties of native meniscus., Objective: In this study, we developed a composite scaffold using decellularized meniscus extracellular matrix (DMECM) and gelatin/chitosan (G/C) to explore a preferable ratio to enhance the elastic modulus and cytotoxicity properties of scaffolds., Methods: The microstructure, porosity, cytotoxicity, and strength of the composite scaffolds were evaluated. The micro-architectures of the samples were evaluated using scanning electron microscope (SEM). Fourier Transform Infrared analysis (FTIR) was used to confirm the chemical structure with different type composite scaffolds. The compressive elastic modulus of all the scaffolds were measured by the universal tensile testing machine DNS300. Calcein-AM (fluorescent green) and propidium iodide (fluorescent red) were used to stain live cells and dead cells. Morphology and spatial distribution of cells within scaffolds were observed by confocal laser scanning microscopy FV 1000., Results: SEM showed that the composite scaffolds had suitable porous structure. CCK-8 and live/dead staining demonstrated that the composite scaffolds had no cytotoxicity and could promote bone marrow mesenchymal stem cells (BMSCs) proliferation. The FTIR results demonstrated the successful mixing of these two elements, and the addition of DMECM improved the elastic modulus and cytotoxicity of G/C composite scaffolds., Conclusions: This study developed a composite scaffold using DMECM and G/C, and demonstrated that it might be suitable for meniscal tissue engineering application.

The increasing rate of injuries to the meniscus indicates the urgent need to develop effective repair strategies. Irreparably damaged menisci can be replaced and meniscus allografts represent the treatment of choice; however, they have several limitations, including availability and compatibility. Another approach is the use of artificial implants but their chondroprotective activities are still not proved clinically. In this situation, tissue engineering offers alternative natural decellularized extracellular matrix (ECM) scaffolds, which have shown biomechanical properties comparable to those of native menisci and are characterized by low immunogenicity and promising regenerative potential. In this article, we present an overview of meniscus decellularization methods and discuss their relative merits. In addition, we comparatively evaluate cell types used to repopulate decellularized scaffolds and analyze the biocompatibility of the existing experimental models. At present, acellular ECM hydrogels, as well as slices and powders, have been explored, which seems to be promising for partial meniscus regeneration. However, their inferior biomechanical properties (compressive and tensile stiffness) compared to natural menisci should be improved. Although an optimal decellularized meniscus scaffold still needs to be developed and thoroughly validated for its regenerative potential in vivo, we believe that decellularized ECM scaffolds are the future biomaterials for successful structural and functional replacement of menisci.

To understand how the complex biomechanical functions of the meniscus are endowed by the nanostructure of its extracellular matrix (ECM), we studied the anisotropy and heterogeneity in the micromechanical properties of the meniscus ECM. We used atomic force microscopy (AFM) to quantify the time-dependent mechanical properties of juvenile bovine meniscus at deformation length scales corresponding to the diameters of collagen fibrils. At this scale, anisotropy in the elastic modulus of the circumferential fibers, the major ECM structural unit, can be attributed to differences in fibril deformation modes: uncrimping when normal to the fiber axis, and laterally constrained compression when parallel to the fiber axis. Heterogeneity among different structural units is mainly associated with their variations in microscale fiber orientation, while heterogeneity across anatomical zones is due to alterations in collagen fibril diameter and alignment at the nanoscale. Unlike the elastic modulus, the time-dependent properties are more homogeneous and isotropic throughout the ECM. These results enable a detailed understanding of the meniscus structure-mechanics at the nanoscale, and can serve as a benchmark for understanding meniscus biomechanical functions, documenting disease progression and designing tissue repair strategies., Statement of Significance: Meniscal damage is a common cause of joint injury, which can lead to the development of post-traumatic osteoarthritis among young adults. Restoration of meniscus function requires repairing its highly heterogeneous and complex extracellular matrix. Employing AFM, this study quantifies the anisotropic and heterogeneous features of the meniscus ECM structure and mechanics. The micromechanical properties are interpreted within the context of the collagen fibril nanostructure and its variation with tissue anatomical locations. These results provide a fundamental structure-mechanics knowledge benchmark, against which, repair and regeneration strategies can be developed and evaluated with respect to the specialized structural and functional complexity of the native tissue., (Copyright © 2017 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.)

Tissue-engineered meniscus regeneration is a very promising treatment strategy for meniscus lesions. However, generating the scaffold presents a huge challenge for meniscus engineering as this has to meet particular biomechanical and biocompatibility requirements. In this study, we utilized acellular meniscus extracellular matrix (AMECM) and demineralized cancellous bone (DCB) to construct three different types of three-dimensional porous meniscus scaffold: AMECM, DCB, and AMECM/DCB, respectively. We tested the scaffolds' physicochemical characteristics and observed their interactions with meniscus fibrochondrocytes to evaluate their cytocompatibility. We implanted the three different types of scaffold into the medial knee menisci of New Zealand rabbits that had undergone total meniscectomy; negative control rabbits received no implants. The reconstructed menisci and corresponding femoral condyle and tibial plateau cartilage were all evaluated at 3 and 6 months (n = 8). The in vitro study demonstrated that the AMECM/DCB scaffold had the most suitable biomechanical properties, as this produced the greatest compressive and tensile strength scores. The AMECM/DCB and AMECM scaffolds facilitated fibrochondrocyte proliferation and the secretion of collagen and glycosaminoglycans (GAGs) more effectively than did the DCB scaffold. The in vivo experiments demonstrated that both the AMECM/DCB and DCB groups had generated neomeniscus at both 3 and 6 months post-implantation, but there was no obvious meniscus regeneration in the AMECM or control groups, so the neomeniscus analysis could not perform on AMECM and control group. At both 3 and 6 months, histological scores were better for regenerated menisci in the AMECM/DCB than in the DCB group, and significantly better for articular cartilage in the AMECM/DCB group compared with the other three groups. Knee MRI scores (Whole-Organ Magnetic Resonance Imaging Scores (WORMS)) were better in the AMECM/DCB group than in the other three groups at both 3 and 6 months. At both 3 and 6 months, RT-PCR demonstrated that aggrecan, Sox9, and collagen II content was significantly higher, and mechanical testing demonstrated greater tensile strength, in the AMECM/DCB group neomenisci compared with the DCB group., (Copyright © 2016 The Authors. Published by Elsevier Ltd.. All rights reserved.)