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Related Experiment Videos

Assessing laser-tissue damage with bioluminescent imaging.

Gerald J Wilmink1, Susan R Opalenik, Joshua T Beckham

  • 1Vanderbilt University, Department of Biomedical Engineering, 5824 Stevenson Center, Nashville, Tennessee 37235, USA.

Journal of Biomedical Optics
|September 13, 2006
PubMed
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A new engineered skin model combined with bioluminescent imaging allows noninvasive study of laser-tissue interactions. This model tracks heat shock protein 70 (hsp70) gene expression, aiding in optimizing laser procedures to minimize tissue damage.

Area of Science:

  • Biomedical Engineering
  • Laser-Tissue Interaction
  • Molecular Biology

Background:

  • Traditional models for studying laser-tissue effects have limitations.
  • A need exists for reproducible, noninvasive models for longitudinal gene expression studies.
  • Optimizing laser parameters requires understanding thermal damage responses.

Purpose of the Study:

  • To develop and validate a novel organotypic raft (engineered skin) model for studying laser-induced thermal damage.
  • To utilize bioluminescent imaging (BLI) for noninvasive, quantitative gene expression analysis.
  • To investigate heat shock protein 70 (hsp70) as a marker for thermal stress in engineered skin.

Main Methods:

  • An organotypic raft model composed of human cells in an extracellular matrix was created.

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  • Cells were transfected with an adenovirus expressing luciferase under the hsp70 promoter.
  • Thermal stress was applied using a CO2 laser, and bioluminescence was monitored using BLI.
  • Gene expression (hsp70 mRNA) and protein levels were analyzed via RT-PCR and ELISA.

    • Main Results:

    • Bioluminescent imaging revealed peak hsp70 expression 4-12 hours post-thermal stress.
    • A minimum laser irradiance of 0.679 Wcm2 activated the hsp70 response.
    • Higher irradiance led to reduced hsp70 response due to tissue ablation, confirmed by histological analysis.
    • Luciferase activity accurately reflected hsp70 protein levels.

    Conclusions:

    • Quantitative BLI in engineered tissue equivalents provides a powerful, noninvasive model for sequential gene expression studies.
    • This model serves as a high-throughput screening platform for laser-tissue interaction research.
    • The developed model aids in optimizing laser parameters to minimize undesirable tissue damage.