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To produce a large-sized regenerative joint by the conjugation of small elements combining artificial bone with tissue engineering cartilage, we made small beta tricalcium phosphate (β-TCP)-hexagonal columns (each side: 2 mm, height: 1 cm) as a substitution of subchondral bone, and attempted to achieve mutual bonding of those columns. A large number of cell sheets was produced from mesenchymal stem cells (MSCs) using NIsopropylacrylamide (NIPPAM)-coated culture dishes. Those cell sheets were dripped onto the lateral surface of a β-TCP-hexagonal column. A pair of the columns was combined side-to-side with each other and were then cultured in osteogenic medium. After 3 weeks of culture, the matrix production-promoting stimulation led to bonding of the two columns, and they were not readily separated by lifting one column using forceps. Not only a pair of those columns, but also three or five, were strongly conjugated, and the columns could be lifted as a single mass. Such in vitro bonding of hexagonal columns of β-TCP utilizing MSCs can support the increase in size of regenerative joints and would enable widening of clinical indications for tissue engineering.

journal of hard tissue biology

Fabrication of Stereotyped Beta-Tricalcium-Phosphate Blocks into a Conjugated Structure using Mesenchymal Stem Cell Sheets Prepared in Temperature-Responsive Culture Dishes