Biocompatibility of a synthetic extracellular matrix on immortalized vocal fold fibroblasts in 3-D culture

In order to promote wound repair and induce tissue regeneration, an engineered hyaluronan (HA) hydrogel – Carbylan GSX, which contains di(thiopropionyl) bishydrazide-modified hyaluronic acid, di(thiopropionyl) bishydrazide-modified gelatin and polyethylene glycol diacrylate – has been developed for extracellular matrix (ECM) defects of the superficial and middle layers of the lamina propria. The purpose of this study was to evaluate the biocompatibility of Carbylan GSX in a previously established immortalized human vocal fold fibroblast (hVFF) cell line prior to human clinical trials. Immortalized hVFF proliferation, viability, apoptosis and transcript analysis for both ECM constituents and inflammatory markers were measured for two-dimensional and three-dimensional (3-D) culture conditions. There were no significant differences in morphology, cell marker protein expression, proliferation, viability and apoptosis of hVFF cultured with Carbylan GSX compared to Matrigel, a commercial 3-D control, after 1 week. Gene expression levels for fibromodulin, transforming growth factor-beta1 and tumor necrosis factor-alpha were similar between Carbylan GSX and Matrigel. Fibronectin, hyaluronidase 1 and cyclooxygenase II expression levels were induced by Carbylan GSX, whereas interleukins 6 and 8, Col I and hyaluronic acid synthase 3 expression levels were decreased by Carbylan GSX. This investigation demonstrates that Carbylan GSX may serve as a natural biomaterial for tissue-engineering of human vocal folds.