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Chondrocytes are utilized to cartilage regeneration by being harvested through enzymatic digestion and expanded by monolayer culture. However, these procedures will cause deterioration and dedifferentiation of the chondrocytes. In addition, scaffolds are often needed to provide the cartilage with mechanical strength and three-dimensional structures. We tried to use diced cartilage prepared using a micro-slicer without digestion, monolayer culture or scaffolds. In this study, an appropriate culture condition to induce the fusion of diced cartilage in vitro and cartilage regeneration in vitro and in vivo was determined to realize a scaffold-free cartilage regeneration. As a result, diced cartilages aggregated when they were cultured more than 5 weeks in the media containing 10% fetal bovine serum (FBS). Diced cartilage cultured for 7 weeks with the media containing 10%, followed by the culture with the media containing insulin-like growth factor-1 for 5 weeks in the ultralow attachment plate showed most prominent cartilage formation both in vitro and in vivo. The volume of regenerated cartilage was 2.14 times larger than that of the original cartilage. These results indicated that large regenerative cartilage from a small amount of cartilage was achieved without deterioration or dedifferentiation.

Establishment of a new technique for the fabrication of regenerative cartilage with a microslicer device to prepare three dimensional diced cartilage