Department of Chemical Engineering, University of Massachusetts Lowell, One University Ave, Lowell, MA, 01854, USA.
Department of Plastics Engineering, University of Massachusetts Lowell, One University Ave, Lowell, MA, 01854, USA.
Sci Rep. 2021 May 7;11(1):9808. doi: 10.1038/s41598-021-89229-w.
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.
生长因子洗脱聚合物系统已被广泛报道可改善细胞和组织的结果；然而，细胞培养条件下实际生长因子浓度的测量是有限的。由于缺乏对半衰期的了解，这一问题更加复杂，阻碍了实时生长因子浓度的确定。在这项工作中，使用酶联免疫吸附测定（ELISA）法测定碱性成纤维细胞生长因子（FGF2）的半衰期。测量了聚电解质多层（PEM）中 FGF2 的释放，并将数据拟合到一个简单的降解模型中，从而可以确定培养时间为 2 至 4 天的 FGF2 浓度。在第一个小时后，在 pH 值为 4 的 PEM 组装中，FGF2 的浓度范围为 2.67ng/mL 至 5.76ng/mL，而在 pH 值为 5 的 PEM 组装中，浓度范围为 0.62ng/mL 至 2.12ng/mL。将 CRL-2352 成纤维细胞培养在 pH 值为 4 和 pH 值为 5 的 PEM 上。2 天后，FGF2 洗脱的 PEM 条件显示出改善的细胞计数和扩散。4 天后，只有 pH 值为 4 的组装条件具有更高的细胞计数，而在 pH 值为 5 的 PEM 组装和没有 FGF2 的 PEM 显示出增加的扩散。总体而言，半衰期模型和细胞培养研究为成纤维细胞增殖提供了最佳浓度范围，并为理解时间依赖性 FGF2 浓度如何影响其他细胞类型提供了框架。
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