Use of optical reporter genes to assess sublethal cellular damage following skin ablation

Gerald J. Wilmink; Susan R. Opalenik; Jeffrey M. Davidson; E. Duco Jansen

doi:10.1117/12.767263

20 February 2008 Use of optical reporter genes to assess sublethal cellular damage following skin ablation

Gerald J. Wilmink, Susan R. Opalenik, Jeffrey M. Davidson, E. Duco Jansen

Proceedings Volume 6854, Optical Interactions with Tissue and Cells XIX; 685403 (2008) https://doi.org/10.1117/12.767263
Event: SPIE BiOS, 2008, San Jose, California, United States

Abstract

Numerous medical procedures utilize pulsed lasers to remove unwanted biological tissue. Mid-infrared wavelengths which preferentially target protein absorption bands ablate tissue more efficiently than wavelengths targeting water absorption. However, the mechanism responsible for this finding has not been established. In this report, we combine optical imaging and conventional techniques to assess lethal and sublethal collateral damage after ablative surgery with a Free Electron Laser (FEL). Heat shock protein expression was used to evaluate tissue damage in a transgenic mouse strain, with the hsp70 promoter driving luciferase and GFP expression (hsp70A1-L2G). To examine wavelength-dependence in the mid-IR, laser surgery was conducted on the hsp70A1-L2G mouse model using wavelengths targeting protein (amide II band, 6.45 μm), both water and protein (amide I band, 6.10 μm); and water (2.94 μm). Hsp70-driven luciferase activity was used as a quantitative biomarker for intracellular damage, and histological analyses were conducted to measure the depth of thermal damage. For all of the wavelengths tested, the bioluminescent data showed that the magnitude of hsp70 expression was dose-dependent. Tissues treated at 6.45 µm had approximately 2x higher hsp70 expression than tissues treated at 6.10 μm. Histology showed that immediate tissue injury at the 6.45 μm wavelength was ~2x deeper than at 6.10 μm. The 6.10 μm wavelength generated the least amount of epidermal hyperplasia. Overall, the data suggests that 6.10 μm is a superior wavelength for cutaneous laser ablation procedures.

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Gerald J. Wilmink, Susan R. Opalenik, Jeffrey M. Davidson, and E. Duco Jansen "Use of optical reporter genes to assess sublethal cellular damage following skin ablation", Proc. SPIE 6854, Optical Interactions with Tissue and Cells XIX, 685403 (20 February 2008); https://doi.org/10.1117/12.767263

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KEYWORDS

Tissues

Proteins

Free electron lasers

Laser ablation

Laser tissue interaction

Skin

Green fluorescent protein

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