Tissue Eng Regen Med.  2018 Dec;15(6):721-733. 10.1007/s13770-018-0145-7.

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

• Iyer K
• Chen Z
• Ganapa T
• Wu BM
• Tawil B
• Linsley CS

• KMID: 2441264
• DOI: http://doi.org/10.1007/s13770-018-0145-7

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.

Keyword

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

MeSH Terms

Cell Movement
Cell Proliferation
Cell Survival
Coculture Techniques
Collagen
Collagen Type I
Drug Therapy
Epithelial Cells
Fibrin
Fibroblasts*
In Vitro Techniques*
Keratinocytes*
Mass Screening
Optical Imaging
Skin
Wound Healing*
Wounds and Injuries*
Collagen
Collagen Type I
Fibrin