Effects of extracellular matrix analogues on primary human fibroblast behavior

Abstract

In vitro cell culture is a vital research tool for cell biology, pharmacology, toxicology, protein production, systems biology and drug discovery. Traditional culturing methods on plastic surfaces do not accurately represent the in vivo environment, and a paradigm shift from two-dimensional to three-dimensional (3-D) experimental techniques is underway. To enable this change, a variety of natural, synthetic and semi-synthetic extracellular matrix (ECM) equivalents have been developed to provide an appropriate cellular microenvironment. We describe herein an investigation of the properties of four commercially available ECM equivalents on the growth and proliferation of primary human tracheal scar fibroblast behavior, both in 3-D and pseudo-3-D conditions. We also compare subcutaneous tissue growth of 3-D encapsulated fibroblasts in vivo in two of these materials, Matrigel ™ and Extracel ™ . The latter shows increased cell proliferation and remodeling of the ECM equivalent. The results provide researchers with a rational basis for selection of a given ECM equivalent based on its biological performance in vitro and in vivo, as well as the practicality of the experimental protocols. Biomaterials that use a customizable glycosaminoglycan-based hydrogel appear to offer the most convenient and flexible system for conducting in vitro research that accurately translates to in vivo physiology needed for tissue engineering.

Response of Fibroblasts to Transforming Growth Factor-β1 on Two-Dimensional and in Three-Dimensional Hyaluronan Hydrogels

“…Previous studies have shown Carbylan-GSX is biocompatible and nontoxic to fibroblasts, and it enhances overall healing and tissue regeneration in in vitro 2D environment and in in vivo animal models. [14][15][16][17] HA is a ubiquitous connective tissue glycosaminoglycan that is a vital element of the ECM, providing tissues with an optimal environment for cell attachment, 14,18 migration, and growth. 15 In addition, HA is an important mediator of the inflammatory cascade, not only recruiting immune cells to the site of wounded tissue, but contributing to granulation tissue formation, 15 amplifying the normal wound-healing response, and promoting tissue regeneration.…”

In Vivo Comparison of Biomimetic Approaches for Tissue Regeneration of the Scarred Vocal Fold

“…16 Specific compositions of an HA-based semisynthetic mimic of the native ECM have been optimized for human fibroblasts. 17 To further build on our previous research demonstrating the effectiveness of HA hydrogels in a prophylactic injury model of the vocal fold lamina propria, we developed a rabbit vocal fold scar model to assess three approaches for tissue regeneration. The purpose of this study was to determine if the biomechanical and biological properties of vocal fold lamina propria scar could be improved with injection of (i) autologous fibroblasts, (ii) a semisynthetic ECM (sECM), or (iii) autologous fibroblasts encapsulated into the sECM when compared to saline-treated controls.…”

Engineering a clinically-useful matrix for cell therapy

“…7,8 This versatile sECM offers user-controllable composition and compliance, to which a variety of growth factors or matricellular proteins may be added. 9,10 The elastic moduli of hydrogels can be tailored to match the cell type to achieve optimal regeneration of a new tissue. 11,12 Design criteria for ECM mimetics.…”