Abstract
Introduction: Idiopathic Pulmonary Fibrosis is a process that involves abnormal cell behaviour and increased tissue stiffness. The aim of this study was to elaborate a stiffened three-dimensional (3D) collagen I matrix to study cultured human cells inside.
Methods: 3D matrices were elaborated with native collagen I and it was non-enzymathically glycated with ribose at different conditions. Matrix stiffness was measured with Athomic Force Microscope and collagen deposition was evaluated by confocal reflexion microscope. Primary fibroblasts were obtained from normal human lungs. Cellular viability in 3D matrices was evaluated by AlamarBlue fluorescence assay and LIVE/DEAD kit at different time points.
Results: It was observed that fluorescence AlamarBlue assay requires more time incubation to detect viability in 3D collagen matrices than in 2D cell cultures. An increased stiffness of 3D matrices was achieved with high concentration of collagen and ribose glycation from the second week. Stiff-variability and morphological changes in collagen I fibers was dependent on the media used for the matrix elaboration and the glycation condition. Cell death was detected in higher ribose concentrations. Fibroblasts showed a better-defined morphology and viability in matrices elaborated with lower ribose concentrations and DMEM media.
Conclusion: The development of this 3D collagen I matrix allows the fibroblast growth inside a modified microenvironment. Cell viability depends on different variables and it dramatically decreases with higher concentrations of ribose. This innovated model could help in the study of cell behaviour and phenotype at different conditions.
Supported by: SEPAR, SOCAP, FUCAP, PS09/01757
- © 2011 ERS
