Mimicking Paracrine TGF\u03b21 Signals during Myofibroblast Differentiation in 3D Collagen Networks

Michael Ansorge,Tilo Pompe,Stephanie Möller,Matthias Schnabelrauch,Marina Chkolnikov,Martin Wilde,Jiranuwat Sapudom,Ulf Anderegg

doi:10.1038/s41598-017-05912-x

Michael Ansorge, Tilo Pompe + Show 6 more

Open Access

https://doi.org/10.1038/s41598-017-05912-x

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Journal: Scientific Reports	Publication Date: Jul 18, 2017
Citations: 21	License type: open-access

Affiliation: Leipzig University, University Hospital Leipzig

Abstract

TGFβ1 is a key regulator for induction of tissue remodeling after dermal wounding. We present a model of paracrine delivery of TGFβ1 for differentiation of dermal fibroblasts based on a fibrillar 3D collagen matrix and embedded TGFβ1 releasing microparticles. We found differentiation into myofibroblasts was achieved in a TGFβ1 dependent manner at much lower doses than systemic delivery. This effect is accounted to the slow and sustained TGFβ1 release mimicking paracrine cell signals.

Highlights

Due to the high level of complexity in vivo and difficult access for high-resolution analytical tools, simplified in vitro models are necessary for in-depth understanding of TGFβ1 signaling and the development of therapeutic strategies
TGFβ1 release from 104 μ-beads into 180 μl of 1 wt-% bovine serum albumin (BSA) in phosphate buffered saline (PBS) was determined over 4 days by enzyme-linked immunosorbent assay (ELISA)
Cytokine delivery during wound healing in vivo is achieved by different cell types like macrophages, platelets and fibroblasts which constantly secrete cytokines into the surrounding tissue[5]

Summary

Introduction

Due to the high level of complexity in vivo and difficult access for high-resolution analytical tools, simplified in vitro models are necessary for in-depth understanding of TGFβ1 signaling and the development of therapeutic strategies Such model systems have to closely mimic the in vivo situation to allow for physiologically relevant results and a reduction of ethically controversial animal studies. In the last decade it became evident that cell culture conditions in a 3D ECM context are needed, but are not provided by standard plastic dish cell culture[7] Such model systems should mimic the soft, fibrillar network characteristics of the ECM, but must enable controlled delivery of mediators and cytokines. Differentiation of dermal fibroblasts into myofibroblasts within the 3D biomimetic matrices was determined by staining of TGFβ1 downstream target Smad2/3, which is phosphorylated and afterwards translocated to the nucleus, and αSMA incorporation into the actin cytoskeleton as well-known marker of contractile myofibroblasts (Fig. 1B and C)[2]

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