Abstract:

Objective The effects of the staurosporine on contraction of self-assembled constructs and extracellular matrix syntheses of goat temporomandibular joint discs were investigated. Methods Goat temporomandibular joint disc cells were isolated and cultured to P3, and 5.5×10 ⁶cells were combined with different concentrations of staurosporine (0, 0.1, 1, 10, 100 nmol·L ^-1) in agarose wells and cultured for one week. The samples were frozen and sectioned. Safranin-O, Picro-sirius red and immunohistochemical staining were performed to observe the distributions of the extracellular matrix and the expression of alpha-smooth muscle actin (α-SMA). Enzyme linked immunosorbent assay (ELISA) and Blyscan kits were utilized to quantitatively detect the contents of type Ⅰ collagen (ColⅠ) and glycosaminoglycans (GAGs). Results Each group of goat temporomandibular joint disc cells in the agarose wells were gathered to self-assemble into a disc-shaped base for 4 hours and then to gradually contract into a round shape. The Picro-sirius red staining was strong and indicated collagen distribution. The Safranin-O staining observed GAGs throughout the entire construct. The expression of ColⅠ was strongly positive in the staurosporine groups; however, the expression of α-SMA was weak. ColⅠ and GAGs contents in the staurosporine groups were greater than that of the control group, especially in the 10 nmol·L ^-1 group (P<0.01). Conclusion Staurosporine has a certain effect on the shrinkage of self-assembled constructs; however, such effect is not prominent. Staurosporine contributes to the construction synthesis of extracellular matrix.

Key words: staurosporine, self-assembled construct, temporomandibular joint disc