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Keratinocyte Migration in a Three-Dimensional In Vitro Wound Healing Model Co-Cultured with Fibroblasts

Tissue Engineering and Regenerative Medicine volume 15, pages 721–733 (2018)Cite this article

Abstract

Background:

Because three-dimensional (3D) models more closely mimic native tissues, one of the goals of 3D in vitro tissue models is to aid in the development and toxicity screening of new drug therapies. In this study, a 3D skin wound healing model comprising of a collagen type I construct with fibrin-filled defects was developed.

Methods:

Optical imaging was used to measure keratinocyte migration in the presence of fibroblasts over 7 days onto the fibrin-filled defects. Additionally, cell viability and growth of fibroblasts and keratinocytes was measured using the alamarBlue® assay and changes in the mechanical stiffness of the 3D construct was monitored using compressive indentation testing.

Results:

Keratinocyte migration rate was significantly increased in the presence of fibroblasts with the cells reaching the center of the defect as early as day 3 in the co-culture constructs compared to day 7 for the control keratinocyte monoculture constructs. Additionally, constructs with the greatest rate of keratinocyte migration had reduced cell growth. When fibroblasts were cultured alone in the wound healing construct, there was a 1.3 to 3.4-fold increase in cell growth and a 1.2 to 1.4-fold increase in cell growth for keratinocyte monocultures. However, co-culture constructs exhibited no significant growth over 7 days. Finally, mechanical testing showed that fibroblasts and keratinocytes had varying effects on matrix stiffness with fibroblasts degrading the constructs while keratinocytes increased the construct’s stiffness.

Conclusion:

This 3D in vitro wound healing model is a step towards developing a mimetic construct that recapitulates the complex microenvironment of healing wounds and could aid in the early studies of novel therapeutics that promote migration and proliferation of epithelial cells.

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Acknowledgements

The authors acknowledge funding support from the National Institute of Arthritis and Musculoskeletal and Skin Diseases of the National Institutes of Health under Award Number R21AR064437. The content is solely the responsibility of the authors and does not necessarily represent the official views of the funding sponsors.

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Affiliations

  1. Department of Bioengineering, University of California, Los Angeles, 420 Westwood Plaza, Room 5121, Engineering V, P.O. Box 951600, Los Angeles, CA, 90095-1600, USA

    Kritika Iyer, Zhuo Chen, Teja Ganapa, Benjamin M. Wu, Bill Tawil & Chase S. Linsley

  2. Division of Advanced Prosthodontics and the Weintraub Center for Reconstructive Biotechnology, School of Dentistry, University of California, Los Angeles, 10833 Le Conte Ave, Los Angeles, CA, 90095, USA

    Benjamin M. Wu

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  1. Kritika Iyer
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  2. Zhuo Chen
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Correspondence to Chase S. Linsley.

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Iyer, K., Chen, Z., Ganapa, T. et al. Keratinocyte Migration in a Three-Dimensional In Vitro Wound Healing Model Co-Cultured with Fibroblasts. Tissue Eng Regen Med 15, 721–733 (2018). https://doi.org/10.1007/s13770-018-0145-7

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  • DOI: https://doi.org/10.1007/s13770-018-0145-7

Keywords

  • Collagen
  • Fibrin
  • In vitro model
  • Cell migration
  • Cell proliferation