Fig. 11

Immunohistochemical staining for evaluation of chondrogenesis- and osteogenesis-related proteins. A Positive staining of Col2, Aggrecan, and Col X was evident in cell-loaded groups, while there were almost no positive cells in the first three groups without BMSCs. The EMF group showed the strongest staining of Col2 and Aggrecan proteins. The Col X expression in EMF group was relatively lower compared to the “HAC + gel + MSC” group. B Osteocalcin (OCN) and Col1 were visualized in the last three groups, and their staining was remarkably stronger compared to other two groups. The number of cells positively expressing OCN or Col1 was relatively higher in the EMF group than those in “HAC + gel” and “HAC + gel + MSC” groups. (C) RNA expression of Col 2, Aggrecan, Col X, OCN and Col 1 in regenerated tissue was measured by RT-PCR. The HAC + gel + MSC + EMF group showed highest level of Col 2, Aggrecan, Col X, OCN and Col 1 relative RNA expression, and significantly higher than any other groups. The magnification of immunohistochemical staining pictures are 3X for upper rows and 30X for lower rows. Data are shown as mean ± standard deviation (n = 5). *p < 0.05, **p < 0.01, ***p < 0.001 compared to blank group