Effects of exogenous IGF-1 delivery on the early expression of IGF-1 signaling molecules by alginate embedded chondrocytes

Cartilage tissue engineering remains a significant challenge for both researchers and clinicians. Many strategic approaches, such as the delivery of growth factors to an in vitro cultured cartilage construct, continue to receive significant attention. However, the effects of delivering exogenous signaling molecules on endogenous signaling pathways within an engineered tissue are not well understood. In order to address this concern, we have investigated how the delivery of insulin-like growth factor-1 (IGF-1, delivered at concentrations of 0, 10, 50, and 100 ng/mL) affects the endogenous expression of IGF-1, its receptor (IGF-1R), and a well known IGF-1 binding protein (IGFBP-3) by articular chondrocytes embedded in alginate hydrogels over 8 days. To the best of our knowledge, this is the first report of delivery effects upon endogenous signal expression in a three-dimensional system relevant to tissue engineering objectives. Results showed significant differences in mRNA expression of IGF-1, IGF-1R, type II collagen, and type I collagen by day 8 between the induced versus noninduced IGF-1 groups. At day 8, the induced IGF-1 groups expressed IGF-1 mRNA four times lower than the 0 ng/mL IGF-1 group. Further, the IGF-1R mRNA expression was five times higher for the groups exposed to exogenous IGF-1 versus the 0 ng/mL IGF-1 case. Interestingly, the expression of IGFBP-3 decreased for all groups. Type II collagen expression was the highest and type I collagen was the lowest for the IGF-1 delivered samples. Finally, the different concentrations of IGF-1 investigated did not demonstrate significantly different trends in mRNA expression levels. Overall, results indicate that exogenous IGF-1 delivery does affect signaling molecule expression by chondrocytes embedded in alginate hydrogels, particularly downregulating the delivered signal while upregulating its receptor.