TUNEL assay for histopathologic evaluation of irreversible chromosomal damage following nonablative fractional photothermolysis

Jordan P. Farkas, James A. Richardson, John E. Hoopman, Spencer A. Brown, Jeffrey M. Kenkel

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)1660-1668
Number of pages9
JournalPlastic and Reconstructive Surgery
Volume122
Issue number6
DOIs
StatePublished - Dec 2008

Fingerprint

Transferases
Dermis
Epidermis
Wounds and Injuries
Lasers
Necrosis
Erbium
Abdominoplasty
Apoptosis
Rejuvenation
Equipment and Supplies
Skin
Phototherapy
DNA Nucleotidylexotransferase
Paraffin
Formaldehyde
deoxyuridine triphosphate
Coloring Agents
Collagen
Hot Temperature

ASJC Scopus subject areas

  • Surgery

Cite this

TUNEL assay for histopathologic evaluation of irreversible chromosomal damage following nonablative fractional photothermolysis. / Farkas, Jordan P.; Richardson, James A.; Hoopman, John E.; Brown, Spencer A.; Kenkel, Jeffrey M.

In: Plastic and Reconstructive Surgery, Vol. 122, No. 6, 12.2008, p. 1660-1668.

Research output: Contribution to journalArticle

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