REGENERATION OF DAMAGED GROWTH PLATE USING IGF-I PLASMID-RELEASING POROUS PLGA SCAFFOLDS

Growth plate injuries account for 15-30% of long bone fractures in children. About 10% of these result in significant growth disturbances due to formation of a boney bar. If not treated correctly, this can lead to life-lasting consequences of limb length inequalities and angular deformities. Current treatments for growth plate injuries include removal of boney bar and insertion of fat, silicone, bone cement, etc.. This treatment y is inadequate, leaving almost half of these patients with continued deformities. This dissertation reports characterization of a DNA–containing porous poly(lactic-co-glycolic acid) (PLGA) scaffold system, chondrogenesis using insulin-like growth factor I (IGF-I) plasmid-releasing scaffolds in vitro, and in vivo testing of IGF-I plasmid-releasing scaffolds to regenerate growth plate . Controlled release of naked and DNA complexed with polyethylenimine (PEI) was achieved from porous PLGA scaffolds. PEI affected release of complexes from PLGA scaffolds, as PEI:DNA complexes were released at a lower rate compared to naked DNA encapsulated in low molecular weight (LMW) and high molecular weight PLGA scaffolds, as well as hydrophilic and hydrophobic PLGA scaffolds. Hydrophilicity and molecular weight of PLGA affected the release profiles of both naked DNA and PEI:DNA complexes from the scaffolds, as evidenced by later peak DNA and PEI:DNA release with increasing hydrophilicity and molecular weight. LMW hydrophilic PLGA scaffolds supported growth and chondrogenic differentiation of mesenchymal multipotent D1 cells, chondrocytes, and bone marrow cells (BMCs) in vitro. Culturing BMCs on IGF-I plasmid-encapsulated scaffolds resulted in elevated expression of IGF-I compared to blank scaffolds. Removal of boney bar and implantation of IGF-I plasmid-releasing LMW PLGA scaffolds in a rabbit model of growth plate injury resulted in some improvement of leg angular deformity compared to no scaffold implantation. Histological analysis of the newly developed cartilage showed growth plate-like columnar arrangement of chondrocytes in a defect that received IGF-I plasmid encapsulated scaffold, although the level of organization of newly formed cartilage was inferior to that of native growth plate. This appears to be the first report of the regeneration of growth plate-like structure without the use of stem cells in an animal model of physeal injury