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In vitro cultured fetal fibroblasts have myofibroblast-associated characteristics and produce a fibrotic-like environment upon stimulation with TGF-β1: Is there a thin line between fetal scarless healing and fibrosis?

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Abstract

Transforming growth factor-β (TGF-β) is a cytokine occurring in three isoforms with an important function in development and wound healing. In wound healing, prolonged TGF-β signaling results in myofibroblast differentiation and fibrosis. In contrast, the developing second-trimester fetal skin contains high levels of all three TGF-β isoforms but still has the intrinsic capacity to heal without scarring. Insight into TGF-β signal transduction during fetal wound healing might lead to methods to control the signaling pathway during adult wound healing. In this study, we imitated wound healing in vitro by stimulating fibroblasts with TGF-β1 and examining myofibroblast differentiation. The aim was to gain insight into TGF-β signaling in human fibroblasts from fetal and adult dermis. First, TGF-β1 stimulation resulted in similar or even more severe upregulation of myofibroblast-associated genes in fetal fibroblasts compared to adult fibroblasts. Second, fetal fibroblasts also had higher protein levels of myofibroblast-marker α-smooth muscle actin (α-SMA). Third, stimulated fetal fibroblasts in collagen matrices had higher protein levels of α-SMA, produced more of the fibrotic protein fibronectin splice-variant extra domain A (FnEDA), and showed enhanced contraction. Finally, fetal fibroblasts also produced significant higher levels of TGF-β1. Altogether, these data show that in vitro cultured fetal fibroblasts have myofibroblast-associated characteristics and do produce a fibrotic environment. As healthy fetal skin has high levels of TGF-β1, FnEDA, and collagen-III as well, these findings correlate with the in vivo situation. Therefore, our study demonstrates that there are similarities between fetal skin development and fibrosis and shows the necessity to discriminate between these processes.

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Acknowledgements

The authors thank J. Braam, BSc., for the work she did during her internship. Furthermore, we are grateful to the personnel of the Centre for Contraception, Sexual Health and Abortion (CASA) in Leiden and of the Rode Kruis Ziekenhuis and the Association of Dutch Burn Centers (ADBC), both in Beverwijk, for collecting and processing tissues. We thank Dr. Ingo Herschel and Dr. Leon Olde Damink from Matricel GmbH for kindly providing Novomaix. This work was supported by the Netherlands Institute of Regenerative Medicine (NIRM, FES0908).

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Authors and Affiliations

  1. Department of Molecular Cell Biology and Immunology (MCBI), VU University Medical Center (VUmc), Zeestraat 27-29, Beverwijk, 1941 AJ, Amsterdam, The Netherlands

    M. Walraven, J. J. Akershoek & R. H. J. Beelen

  2. Association of Dutch Burn Centres (ADBC), Zeestraat 27-29, Beverwijk, 1941 AJ, Amsterdam, The Netherlands

    M. Walraven, J. J. Akershoek & M. M. W. Ulrich

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  1. M. Walraven
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  2. J. J. Akershoek
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  3. R. H. J. Beelen
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  4. M. M. W. Ulrich
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Correspondence to M. M. W. Ulrich.

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This article does not contain any studies with human participants or animals performed by any of the authors.

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Informed consent was given for all donated tissues and collection was done according to national guidelines formulated by the federation of Dutch medical scientific societies (FMWV/FDMSS).

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Walraven, M., Akershoek, J.J., Beelen, R.H.J. et al. In vitro cultured fetal fibroblasts have myofibroblast-associated characteristics and produce a fibrotic-like environment upon stimulation with TGF-β1: Is there a thin line between fetal scarless healing and fibrosis?. Arch Dermatol Res 309, 111–121 (2017). https://doi.org/10.1007/s00403-016-1710-3

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  • DOI: https://doi.org/10.1007/s00403-016-1710-3

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