Despite many outstanding research works on cartilage tissue engineering, actual
clinical application is not quite successful because of the absorption and progressive distortion of
tissue engineered cartilage. We have developed a new method of cartilage tissue engineering
comprising chondrocyte mixed Pluronic F-127 and cultured chondrocyte cell sheet which entirely
cover the cell-Pluronic complex. We believe the addition of cultured chondrocyte cell sheet
enhances the efficacy of chondrogenesis in vivo.
Human ear cartilage piece was enzymatically dissociated and chondrocyte suspension was
acquired. Chondrocytes were cultured and expanded as the routine manner. Cultured chondrocytes
were plated in high-density monolayer and cultured with Chondrogenic media in 5% CO2 incubator.
After 3 weeks of culture, chondrocyte cell sheet was formed and complete single sheet of
chondrocyte could be harvested by gentle manipulation of culture plate with a cell scraper.
Chondrocyte-Pluronic complex was established by mixing 1x 106 cells with 0.5 of Pluronic F-
127. Chondrocyte-Pluronic complex was completely covered with a sheet of cultured chondrocyte.
The completed tissue engineered constructs were implanted into the subcutaneous tissue pocket of
nude mice on the back. Tissue engineered constructs without cultured cell sheet were used as
control. Samples were harvested at 8 weeks postoperatively and they were subjected to histological
analysis and assayed for glycosaminoglycan (GAG), and type II collagen. Grossly, the size of
cartilage specimen of cultured chondrocyte cell sheet covered group was larger than that of the
control. On histologic examination, the specimen of cultured chondrocyte cell sheet covered group
showed lacunae-containing cells embedded in a basophilic matrix. The chondrocyte cell sheet
covered group specimen resembled mature or immature cartilage. The result of measurement of
GAG and type II collagen of cartilage specimen of cultured chondrocyte sheet covered group was
higher than that of the control.
In conclusion, the new method of cartilage tissue engineering using chondrocyte cell sheet seems
to be an effective method providing higher cartilage tissue gain and reliable success rate for
cartilage tissue engineering.