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Related Experiment Videos

Modeling aberrant wound healing using tissue-engineered skin constructs and multiphoton microscopy.

Behrooz A Torkian1, Alvin T Yeh, Rodney Engel

  • 1Department of Otolaryngology- Head and Neck Surgery, University of California, Irvine Medical Center, Irvine, CA 92612, USA.

Archives of Facial Plastic Surgery
|May 19, 2004
PubMed
Summary

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This study introduces a new wound healing model using multiphoton microscopy (MPM) to observe keloid fibroblast activity. Keloid fibroblasts deposit collagen faster than normal fibroblasts in engineered skin, aiding scar research.

Area of Science:

  • Biomedical Engineering
  • Tissue Engineering
  • Dermatology

Background:

  • Keloids and hypertrophic scars arise from abnormal wound healing, a common complication of injury or surgery.
  • Previous research on aberrant wound healing was limited by in vitro methods requiring cell destruction.
  • Multiphoton microscopy (MPM) offers a dye-free imaging technique using endogenous tissue chromophores like collagen and NADH.

Purpose of the Study:

  • To introduce a novel wound-healing model for investigating keloid-derived fibroblast activity and collagen production.
  • To analyze fibroblast behavior and extracellular matrix changes in intact, engineered tissue constructs over time.

Main Methods:

  • Engineered skin constructs (RAFTs) were created with keloid or normal fibroblasts in collagen gel and a keratinocyte layer.

Related Experiment Videos

  • Multiphoton microscopy (MPM) with second-harmonic generation (SHG) was used to image collagen in intact constructs.
  • Constructs were wounded, and serial MPM and phase-contrast images were captured over 8 days to track extracellular matrix remodeling.

    • Main Results:

    • The RAFT constructs successfully mimicked human skin's bilayer structure.
    • MPM SHG imaging revealed increased collagen deposition post-wounding.
    • Constructs with keloid fibroblasts exhibited a higher rate of collagen deposition compared to those with normal fibroblasts.

    Conclusions:

    • The developed MPM model allows for serial, non-destructive observation of intact tissue specimens.
    • This model validates the biological activity of RAFT constructs in simulating wound healing processes.
    • The findings highlight distinct collagen deposition rates between keloid and normal fibroblasts in a dynamic wound model.