Journal: Acta biomaterialia

Article Title: Functionally graded multilayer scaffolds for in vivo osteochondral tissue engineering

doi: 10.1016/j.actbio.2018.07.039

Figure Lengend Snippet: The cell-laden top and middle layers support in vivo formation of stratified cartilage with lubricin-rich surface. Alcian Blue staining and immunohistochemical staining for type II collagen and lubricin of the cell-laden trilayer scaffold at 8 weeks post-implantation. Scale bars represent 200 µm. High magnification images for top, middle, and bottom layers of the trilayer scaffold as well as native osteochondral tissue are also shown. Scale bars indicate 50 µm. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: The treated sections were immersed in a blocking solution containing 3% (w/v) normal goat serum and 0.1% (v/v) Triton X-100 at 25 °C for 1 h and incubated with primary antibodies against type I collagen (1:100, rabbit polyclonal, Fitzerald, 70R-CR007x), type II collagen (1:50, rabbit polyclonal, Fitzerald, 70R-CR008), type X collagen (1:50, rabbit polyclonal, Fitzerald, 20R-CR030), osteocalcin (1:100, rabbit polyclonal, Abcam, ab93876), lubricin (1:50, goat polyclonal, Santa Cruz Biotechnology, sc-50079), or lamin B (1:100, rabbit polyclonal, Thermo Scientific, RB10569-P0) in a blocking solution at 4 °C for 16 h. The sections were washed with PBS and treated with 3% (v/v) hydrogen peroxide for 7 min to deactivate endogenous peroxidase.

Techniques: In Vivo, Staining, Immunohistochemical staining