Abstract
Growth factor-eluting polymer systems have been widely reported to improve cell and tissue outcomes; however, measurements of actual growth factor concentration in cell culture conditions are limited. The problem is compounded by a lack of knowledge of growth factor half-lives, which impedes efforts to determine real-time growth factor concentrations. In this work, the half-life of basic fibroblast growth factor (FGF2) was determined using enzyme linked immunosorbent assay (ELISA). FGF2 release from polyelectrolyte multilayers (PEMs) was measured and the data was fit to a simple degradation model, allowing for the determination of FGF2 concentrations between 2 and 4 days of culture time. After the first hour, the FGF2 concentration for PEMs assembled at pH = 4 ranged from 2.67 ng/mL to 5.76 ng/mL, while for PEMs assembled at pH = 5, the concentration ranged from 0.62 ng/mL to 2.12 ng/mL. CRL-2352 fibroblasts were cultured on PEMs assembled at pH = 4 and pH = 5. After 2 days, the FGF2-eluting PEM conditions showed improved cell count and spreading. After 4 days, only the pH = 4 assembly condition had higher cells counts, while the PEM assembled at pH = 5 and PEM with no FGF2 showed increased spreading. Overall, the half-life model and cell culture study provide optimal concentration ranges for fibroblast proliferation and a framework for understanding how temporal FGF2 concentration may affect other cell types.
Highlights
Growth factor-eluting polymer systems have been widely reported to improve cell and tissue outcomes; measurements of actual growth factor concentration in cell culture conditions are limited
While the half-life model developed in this work was applied to a specific polyelectrolyte multilayers (PEMs), the model can be applied to any culture environment where FGF2 vs. time data is available, including traditional culture systems and those containing biomaterials capable of controlled release
Since the model is based on enzyme linked immunosorbent assay (ELISA)-detectable concentrations of FGF2, the half-life of FGF2 was determined at concentrations that are relevant to culture of cells such as fibroblasts and myoblasts
Summary
Growth factor-eluting polymer systems have been widely reported to improve cell and tissue outcomes; measurements of actual growth factor concentration in cell culture conditions are limited. The structure and properties of the resulting material depend on factors including the assembly method, polyelectrolyte concentrations, pH, and temperature[3,13,14,15] For this reason, polyelectrolytes have been used for a range of cell scaffolds and surface coatings capable of releasing specific nanoparticles, pharmaceuticals, and proteins[7,8,9,10,11,16]. Methods for measuring GF concentration include enzyme-linked immunosorbent assay (ELISA), fluorescent labeling, circular dichroism, and western blot[3,4,5,17,26,27,28,29]. Generating an accurate model of FGF2 degradation using ELISA-derived results necessitates determining its ELISA-detectable half-life
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