Cell-based cartilaginous graft as active PGRN delivery system.

• Main Author: Tan, Li Fang.
• Other Authors: Wang Dongan
• Format: Final Year Project
• Language: English
• Published: 2013
• Subjects: DRNTU::Engineering
• Online Access: http://hdl.handle.net/10356/52713
• Institution: Nanyang Technological University

Arthritis is a group of chronic diseases distinguished by the inflammation of joints, due to the proteolytic degradation of extracellular matrix components, which leads to the gradual loss of articular cartilage and bone. The most frequently-occurring arthritis is osteoarthritis (OA), where injury or trauma results in the degeneration of the joint. Another form, the rheumatoid arthritis (RA), happens when the immune cells within the body wrongly attack healthy tissue, damaging the joint and resulting in a systemic autoimmune disorder. With native articular hyaline cartilage being avascular and aneural, most patients do not notice initial damage to the cartilage. The injuries then progress to become more generic, but cartilage loss extending to the subchondral bone is permanent. The problems arising from arthritic conditions are expected to grow as longevity of the general world’s population increases. Progranulin (PGRN), an effective TNFα antagonist, has been established to exert anti-inflammatory effects and protect cartilage from macrophages secreting TNFα. Unlike current therapies approved for RA treatment that mainly target cytokines such as TNFα, PGRN binds to TNF receptors (TNFR), inhibiting TNFα-TNFR interactions and the following cascade of inflammatory reactions. Despite the success of current RA treatment in improving the quality of life in some patients, these therapies with TNFα inhibitors remain ineffective for up to 50% of them; suggesting that PGRN, with its different mechanism adopted, may work. In this study, we established a recombinant pAdeno-X DNA plasmid containing the PGRN gene of interest. These adenoviral vectors were then expanded in HEK 293 cells, before purification of the virus to transfect chondrocytes in both 2D and 3D constructs. Analysis showed good cell viability following transfection, as well as a positive PGRN secretion profile, suggesting its potential as future therapeutic treatment for arthritic conditions.