Controlling Fibroblast Proliferation with Dimensionality-Specific Response by Stiffness of Injectable Gelatin Hydrogels

Abstract

We report an injectable hydrogel system with tunable stiffness for controlling the proliferation rate of human fibroblasts (HFF-1) in both two-dimensional (2D) and three-dimensional (3D) culture environments for potential use as a wound dressing material. The hydrogel composed of gelatin–hydroxyphenylpropionic acid (Gtn–HPA) conjugate was formed by the oxidative coupling of HPA moieties catalyzed by hydrogen peroxide (H2O2) and horseradish peroxidase (HRP). The stiffness of the hydrogels was controlled well by varying the H2O2 concentration. The effects of hydrogel stiffness on the proliferation rate of HFF-1 in both 2D and 3D were investigated. We found that the proliferation rate of HFF-1 using Gtn–HPA hydrogels was strongly dependent on the hydrogel stiffness, with a dimensionality-specific response. In the 2D studies, the HFF-1 exhibited a higher proliferation rate when the stiffness of the hydrogel was increased. In contrast, the HFF-1 cultured inside the hydrogel remained non-proliferative for 12 days before a stiffness-dependent proliferation profile was shown. The proliferation rate decreased with an increase in stiffness of the hydrogel in a 3D culture environment, unlike in a 2D environment.