Cartilage tissue engineering on macroporous scaffolds using human tooth germ stem cells.

The main objective was to study cartilage regeneration through differentiation of human tooth germ stem cells (HTGSCs) into chondrocytes on different three-dimensional (3D) scaffolds (PCL, PLLA and PCL–PLLA). Scaffold topographies were studied by scanning electron microscopy and it was found that the scaffolds had interconnected macroporous structures. HTGSCs were isolated from impacted third molar tooth germs of young adult patients and grown for 3 weeks on the scaffolds in chondrogenic differentiation medium. Cell proliferation on the scaffolds was determined by MTS assay and it was observed that all scaffolds supported cell proliferation. Immunostaining was carried out for morphological and differentiation analyses. Immunohistochemical analyses revealed that the cells attached onto the scaffolds and deposited cartilage-specific extracellular matrix (ECM). Real-time PCR was performed to determine the expression levels of cartilage-specific genes. After 21 days of incubation in cartilage differentiation medium, expression of collagen type II increased only in the cells seeded onto PCL–PLLA blend scaffolds. Similarly, aggrecan expression was the highest on PCL–PLLA scaffolds after 3 weeks. These results suggest that all the scaffolds, and especially PCL–PLLA, were suitable for chondrogenic differentiation of HTGSCs. Copyright © 2014 John Wiley & Sons, Ltd.