Quantitative Response Measurement of Cell Substrate Interactions via RT-PCR
High-throughput metrologies for the rapid and systematic evaluation of synthetic materials, which would elucidate a candidate’s potential biocompatibility, are needed. New synthetic methodologies have enabled a remarkable advance in the rational design of polymeric materials that actively control cellular and physiologic responses for use in tissue engineering applications. These methods, which afford precise control over molecular architecture, mass, and composition, produce well-defined materials that are being incorporated into scaffolds capable of supporting and regulating the adhesion, growth, and function of target cells while being minimally detrimental to normal cellular processes and surrounding tissues. A method to assess the biocompatibility of materials in vitro using real-time polymerase chain reaction (RT-PCR) has been developed within the Biomaterials group. Inflammatory responses play a prominent role in the biocompatibility of materials, as indicated by the induction of the cytokines interleukin-1 beta (IL-1b ) and tumor necrosis factor- alpha (TNF-a ). RT-PCR measurements have quantitatively assessed the genetic expression profiles for these cytokines in response to tissue-engineered scaffolds, copolymer blends, and functionalized nanoparticles. The further development of additional marker to measure cellular responses to biomaterials at the genetic level is of great importance. The quantification of critical cellular responses to interactions with synthetic substrates by RT-PCR will be described.