Ann Dermatol.  2015 Jun;27(3):283-290. 10.5021/ad.2015.27.3.283.

Usefulness of Skin Explants for Histologic Analysis after Fractional Photothermolysis

• Park GH
• Chang SE
• Bang S
• Won KH
• Won CH
• Lee MW
• Choi JH
• Moon KC

• KMID: 2352499
• DOI: http://doi.org/10.5021/ad.2015.27.3.283

Abstract

BACKGROUND
Fractional laser resurfacing treatment has been extensively investigated and is widely used. However, the mechanism underlying its effects is poorly understood because of the ethical and cosmetic problems of obtaining skin biopsies required to study the changes after laser treatment.
OBJECTIVE
To evaluate the usefulness of human skin explants for the investigation of fractional photothermolysis.
METHODS
Full-thickness discarded skin was treated in 4 ways: no treatment (control), fractional carbon dioxide laser, fractional Er:YAG laser, and fractional 1,550-nm erbium-doped fiber laser. Both treated and non-treated skin samples were cultured ex vivo at the air-medium interface for 7 days. Frozen tissue was sectioned and stained with hematoxylin & eosin for histologic examination and nitro blue tetrazolium chloride for viability testing.
RESULTS
Skin explants cultured for up to 3 days exhibited histologic changes similar to those observed in in vivo studies, including microscopic treatment zones surrounded by a thermal coagulation zone, re-epithelialization, and formation of microscopic epidermal necrotic debris. However, the explant structure lost its original form within 7 days of culture. The viability of skin explants was maintained for 3 days of culture but was also lost within 7 days.
CONCLUSION
The skin explant model may be a useful tool for investigating the immediate or early changes following fractional photothermolysis, but further improvements are required to evaluate the long-term and dermal changes.

Keyword

Tissue culture techniques; Tissue survival; Laser therapy; Cosmetic techniques

MeSH Terms

Biopsy
Cosmetic Techniques
Eosine Yellowish-(YS)
Hematoxylin
Humans
Laser Therapy
Lasers, Gas
Re-Epithelialization
Skin*
Tissue Culture Techniques
Tissue Survival
Eosine Yellowish-(YS)
Hematoxylin

Figure

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  Fig. 1 Ex vivo organ culture of skin explants. Method (A): culture method in which the sample is incubated in medium with the apical epithelial surface up at the air-medium interface. Method (B): culture method using Transwell chambers and microporous inserts filled with 3% agarose.

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  Fig. 2 Histologic appearance of human skin explants cultured using culture method (A) (H&E, ×100). NAFR: fractional nonablative laser treatment, Er:YAG FR: fractional Er:YAG laser treatment, CO2 FR: fractional carbon dioxide laser treatment.

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  Fig. 3 Histologic appearance of human skin explants cultured using culture method (B) (H&E, ×100). NAFR: fractional nonablative laser treatment, Er:YAG FR: fractional Er:YAG laser treatment, CO2 FR: fractional carbon dioxide laser treatment.

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  Fig. 4 Viability of human skin explants cultured using culture method (A) (nitro blue tetrazolium chloride, ×100). NAFR: fractional nonablative laser treatment, Er:YAG FR: fractional Er:YAG laser treatment, CO2 FR: fractional carbon dioxide laser treatment.

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  Fig. 5 Viability of human skin explants cultured using culture method (B) (nitro blue tetrazolium chloride, ×100). NAFR: fractional nonablative laser treatment, Er:YAG FR: fractional Er:YAG laser treatment, CO2 FR: fractional carbon dioxide laser treatment.

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