Abstract
A continuing challenge in cartilage tissue engineering for cartilage regeneration is the creation of a suitable synthetic microenvironment for chondrocytes and tissue regeneration. The aim of this study was to develop a highly tunable hybrid scaffold based on a silk fibroin matrix (SM) and a hyaluronic acid (HA) hydrogel. Human articular chondrocytes were embedded in a porous 3-dimensional SM, before infiltration with tyramine modified HA hydrogel. Scaffolds were cultured in chondropermissive medium with and without TGF-β1. Cell viability and cell distribution were assessed using CellTiter-Blue assay and Live/Dead staining. Chondrogenic marker expression was detected using qPCR. Biosynthesis of matrix compounds was analyzed by dimethylmethylene blue assay and immuno-histology. Differences in biomaterial stiffness and stress relaxation were characterized using a one-step unconfined compression test. Cell morphology was investigated by scanning electron microscopy. Hybrid scaffold revealed superior chondro-inductive and biomechanical properties compared to sole SM. The presence of HA and TGF-β1 increased chondrogenic marker gene expression and matrix deposition. Hybrid scaffolds offer cytocompatible and highly tunable properties as cell-carrier systems, as well as favorable biomechanical properties.
Highlights
Articular cartilage regeneration is still challenging due to its limited intrinsic selfregenerative capacity [1,2]
The HRP concentration had a minor influence on the viscoelastic properties, higher HRP concentration accelerated gelation
The staining is based on an acidophilic metachromatic dye which selectively stains acidic tissue components such as Sulphated glycosaminoglycan (sGAG), which results in a color change from blue to purple
Summary
Articular cartilage regeneration is still challenging due to its limited intrinsic selfregenerative capacity [1,2]. Third-generation ACI products provide a three-dimensional (3D) cell-carrier matrix, which is either composed of a hydrogel made of natural biopolymers such as agarose–alginate, hyaluronic acid (HA), type 1 collagen, or macroporous scaffolds made of natural materials such as type 1/3 collagen, HA, HA/fibrin or synthetic polymers (e.g., polyglycolic acid and poly(lactic-coglycolic)acid) [5,6,7]. Each biomaterial can provide individual advantages for cell viability and differentiation, biomaterials composed of a sole biomaterial are often limited in their overall function. Material stiffness and associated resistance to biomechanical forces, osmotic capacity, degradation behavior and matrix retention have been proven to significantly guide cell differentiation [8,9,10]
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