Abstract
A lot has been invested into understanding how to assemble cartilage tissue in vitro and various designs have been developed to manufacture cartilage tissue with native-like biological properties. So far, no satisfactory design has been presented. Bovine primary chondrocytes are used to self-assemble scaffold-free constructs to investigate whether mechanical loading by centrifugal force would be useful in manufacturing cartilage tissue in vitro. Six million chondrocytes were laid on top of defatted bone disks placed inside an agarose well in 50-ml culture tubes. The constructs were centrifuged once or three times per day for 15 min at a centrifugal force of 771×g for up to 4 weeks. Control samples were cultured under the same conditions without exposure to centrifugation. The samples were analysed by (immuno)histochemistry, Fourier transform infrared imaging, micro-computed tomography, biochemical and gene expression analyses. Biomechanical testing was also performed. The centrifuged tissues had a more even surface covering a larger area of the bone disk. Fourier transform infrared imaging analysis indicated a higher concentration of collagen in the top and bottom edges in some of the centrifuged samples. Glycosaminoglycan contents increased along the culture, while collagen content remained at a rather constant level. Aggrecan and procollagen α1(II) gene expression levels had no significant differences, while procollagen α2(I) levels were increased significantly. Biomechanical analyses did not reveal remarkable changes. The centrifugation regimes lead to more uniform tissue constructs, whereas improved biological properties of the native tissue could not be obtained by centrifugation.
Highlights
Articular cartilage has a limited ability for self-repair and, when damaged, it has a high risk of progressing into osteoarthritis, if not intervened
There are still several drawbacks with all of these methods, which have led to attempts to develop various tissue engineering techniques to create in vitro grown implants in hopes of combining the advantages of autologous chondrocyte implantation and mosaicplasty
It is relatively easy to produce a threedimensional construct out of chondrocytes but deciphering ways to improve the structure of the extracellular matrix (ECM) and the orientational assembly of its components is more complicated
Summary
Articular cartilage has a limited ability for self-repair and, when damaged, it has a high risk of progressing into osteoarthritis, if not intervened. There are still several drawbacks with all of these methods, which have led to attempts to develop various tissue engineering techniques to create in vitro grown implants in hopes of combining the advantages of autologous chondrocyte implantation and mosaicplasty. One such approach is to manufacture constructs with the mechanical and biological properties of the native articular cartilage (Moutos et al 2007). Advances in the development of cell-laden hydrogels still offer new possibilities to improve the physical and biological properties of the next-generation matrices applicable for osteochondral/cartilage tissue engineering. Scaffold-free approaches can mimic elements of native developmental processes (DuRaine et al 2015)
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