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TUNEL Assay for Histopathologic Evaluation of Irreversible Chromosomal Damage following Nonablative Fractional Photothermolysis

Farkas, Jordan P. M.D.; Richardson, James A. D.V.M., Ph.D.; Hoopman, John E. C.M.L.S.O.; Brown, Spencer A. Ph.D.; Kenkel, Jeffrey M. M.D.

Plastic and Reconstructive Surgery: December 2008 - Volume 122 - Issue 6 - p 1660-1668
doi: 10.1097/PRS.0b013e31818a9a26
Experimental: Original Articles

Background: Fractional photothermolysis is extremely popular in skin rejuvenation and remodeling procedures. However, the extent of thermal cellular injury beyond the borders of the coagulated microcolumns produced with fractional phototherapy is undefined.

Methods: Six abdominoplasty patients were pretreated with the Lux1540 Fractional Erbium device (Palomar, Inc., Burlington, Mass.) at various clinical laser settings. After tissue excision, the panni were immediately biopsied. Biopsy specimens were fixed in formalin, embedded in paraffin, sectioned, and evaluated with the terminal deoxynucleotidyl transferase–mediated deoxyuridine triphosphate nick end labeling (TUNEL) procedure for cellular necrosis/apoptosis. Tissue was sectioned horizontally and longitudinally to help define the depth and distribution of the microcolumns of injury in a three-dimensional plane.

Results: The extent of cellular necrosis/apoptosis at variable depths within the epidermis and dermis was demonstrated successfully with the TUNEL technique. After the Lux1540 treatment, TUNEL-positive nuclei were identified in a vertically oriented fashion that extended from the epidermis into the papillary and reticular dermis, highlighting the areas of injury. The TUNEL-positive nuclei defined lesions that were approximately 175 to 225 μm in diameter and penetrated to variable depths (200 to 900 μm), depending on the fluence used for treatment (18 to 100 mJ).

Conclusions: TUNEL immunofluorescent labeling provided an accurate assessment of cellular damage within and surrounding the microthermal zones of coagulated collagen with respect to column depth and width. Because of its specificity, the TUNEL assay can be a useful adjunct to other histologic stains used to characterize cellular damage and matrix denaturation in skin treated with any fractional ablative or nonablative laser device.

Dallas, Texas

From the Department of Plastic Surgery, Clinical Center for Cosmetic Laser Treatment, and the Departments of Pathology and Environmental Health and Safety, University of Texas Southwestern Medical Center at Dallas.

Received for publication January 30, 2008; accepted May 29, 2008.

Disclosure: The research grant and devices used in the study were provided by Palomar Medical Technologies.

Jeffrey M. Kenkel, M.D.; Department of Plastic Surgery; Clinical Center for Cosmetic Laser Treatment; University of Texas Southwestern Medical Center at Dallas; 1801 Inwood Road; Dallas, Texas 75390-9163;

©2008American Society of Plastic Surgeons