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Volume 12
Issue 3
10.3390/cosmetics12030111
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This is an early access version, the complete PDF, HTML, and XML versions will be available soon.
Article

Two-Photon Microscopy for the Investigation of Morphological and Quantitative Changes in Skin Chrono- and Photo-Aging

by
Stefano Bighetti
1,*,
Chiara Rovati
1,
Luca Bettolini
1,
Mariachiara Arisi
1,
Mariateresa Rossi
1,
Cosetta Ravelli
2,
Sara Rovaris
1,
Marina Venturini
1,
Stefania Mitola
2 and
Piergiacomo Calzavara-Pinton
1
1
Dermatology Department, University of Brescia, ASST Spedali Civili di Brescia, 25123 Brescia, Italy
2
Department of Molecular and Translational Medicine, University of Brescia, 25123 Brescia, Italy
*
Author to whom correspondence should be addressed.
Cosmetics 2025, 12(3), 111; https://doi.org/10.3390/cosmetics12030111
Submission received: 14 March 2025 / Revised: 19 May 2025 / Accepted: 23 May 2025 / Published: 26 May 2025
(This article belongs to the Special Issue Feature Papers in Cosmetics in 2025)
Versions Notes

Abstract

Skin aging is influenced both by intrinsic factors and environmental exposures, such as UV radiation, which accelerate structural changes within the skin’s extracellular matrix (ECM). Understanding these changes is crucial for developing effective anti-aging treatments. This pilot cross-sectional study examined skin biopsy samples from three Caucasian male subjects with different levels of UV exposure, aiming to evaluate the effectiveness of two-photon microscopy (2PM) and Second Harmonic Generation (SHG) in visualizing and quantifying structural changes associated with skin aging. The samples were analyzed using 2PM to assess the structure and density of collagen and elastin fibers within the ECM. Integrated optical density (IOD) and the SHG-to-Autofluorescence Aging Index of the Dermis (SAAID) were used for quantitative analysis. This study revealed a significant decrease in collagen density and increased disorganization in the ECM with age. Photo-exposed skin showed a more pronounced degradation of collagen and a higher increase in elastin content compared to non-photo-exposed skin. The average IOD for collagen was notably lower in elderly subjects compared to younger subjects, with a marked decrease in chronically photo-exposed skin. The SAAID values indicated a substantial impact of photoaging, with lower scores in photo-exposed elderly skin compared to non-exposed skin. In conclusion, 2PM and SHG microscopy were effective in visualizing and quantifying age- and UV-induced skin remodeling, providing valuable insights into the distinct mechanisms driving intrinsic and extrinsic aging.
Keywords: two-photon microscopy; photoaging; extracellular matrix; collagen degradation; aging skin two-photon microscopy; photoaging; extracellular matrix; collagen degradation; aging skin

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MDPI and ACS Style

Bighetti, S.; Rovati, C.; Bettolini, L.; Arisi, M.; Rossi, M.; Ravelli, C.; Rovaris, S.; Venturini, M.; Mitola, S.; Calzavara-Pinton, P. Two-Photon Microscopy for the Investigation of Morphological and Quantitative Changes in Skin Chrono- and Photo-Aging. Cosmetics 2025, 12, 111. https://doi.org/10.3390/cosmetics12030111

AMA Style

Bighetti S, Rovati C, Bettolini L, Arisi M, Rossi M, Ravelli C, Rovaris S, Venturini M, Mitola S, Calzavara-Pinton P. Two-Photon Microscopy for the Investigation of Morphological and Quantitative Changes in Skin Chrono- and Photo-Aging. Cosmetics. 2025; 12(3):111. https://doi.org/10.3390/cosmetics12030111

Chicago/Turabian Style

Bighetti, Stefano, Chiara Rovati, Luca Bettolini, Mariachiara Arisi, Mariateresa Rossi, Cosetta Ravelli, Sara Rovaris, Marina Venturini, Stefania Mitola, and Piergiacomo Calzavara-Pinton. 2025. "Two-Photon Microscopy for the Investigation of Morphological and Quantitative Changes in Skin Chrono- and Photo-Aging" Cosmetics 12, no. 3: 111. https://doi.org/10.3390/cosmetics12030111

APA Style

Bighetti, S., Rovati, C., Bettolini, L., Arisi, M., Rossi, M., Ravelli, C., Rovaris, S., Venturini, M., Mitola, S., & Calzavara-Pinton, P. (2025). Two-Photon Microscopy for the Investigation of Morphological and Quantitative Changes in Skin Chrono- and Photo-Aging. Cosmetics, 12(3), 111. https://doi.org/10.3390/cosmetics12030111

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