| Description |
The aim of this project was to culture CRISPR-modulated cells onto collagen scaffolds in 3D to develop tissue comparable to tendon enthesis and observe whether modulated properties were demonstrated. To do so, modulated stem cells were cultured on 3D collagen gels and histology was performed at various time points. Both edited cell lines showed success by demonstrating proteoglycan deposits and osteogenesis differentiation in 3D culture. Despite observable improvements in these properties, cells failed to penetrate the collagen scaffolds and remained concentrated along the outer layer. A cell seeding device was also successfully designed and used to culture varying ratios of cells to create a tissue gradient. The cultured gradient successfully led to a comparable density gradient along the scaffold. The method of gene modification used in this project is valuable for future use, but improvements must be made for to the scaffold to reach tissue formation comparable to native bone interfaces. This experiment is a critical step in the utilization of tissue engineering to improve both understanding and treatment of degeneration or injury to tendon enthesis and other bone interfaces. |
