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

Introduction: 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. Materials and M...

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Main Authors: Stefano Bighetti, Chiara Rovati, Luca Bettolini, Mariachiara Arisi, Mariateresa Rossi, Cosetta Ravelli, Sara Rovaris, Marina Venturini, Stefania Mitola, Piergiacomo Calzavara-Pinton
Format: Article
Language:English
Published: MDPI AG 2025-05-01
Series:Cosmetics
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Online Access:https://www.mdpi.com/2079-9284/12/3/111
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author Stefano Bighetti
Chiara Rovati
Luca Bettolini
Mariachiara Arisi
Mariateresa Rossi
Cosetta Ravelli
Sara Rovaris
Marina Venturini
Stefania Mitola
Piergiacomo Calzavara-Pinton
author_facet Stefano Bighetti
Chiara Rovati
Luca Bettolini
Mariachiara Arisi
Mariateresa Rossi
Cosetta Ravelli
Sara Rovaris
Marina Venturini
Stefania Mitola
Piergiacomo Calzavara-Pinton
author_sort Stefano Bighetti
collection DOAJ
description Introduction: 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. Materials and Methods: 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. Results: 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. Discussion: The SAAID values indicated a substantial impact of photoaging, with lower scores in photo-exposed elderly skin compared to non-exposed skin. Conclusions: 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.
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spelling doaj-art-b68e3af278ef4287bd9d2cb3d5c037992025-08-20T03:26:52ZengMDPI AGCosmetics2079-92842025-05-0112311110.3390/cosmetics12030111Two-Photon Microscopy for the Investigation of Morphological and Quantitative Changes in Skin Chrono- and Photo-AgingStefano Bighetti0Chiara Rovati1Luca Bettolini2Mariachiara Arisi3Mariateresa Rossi4Cosetta Ravelli5Sara Rovaris6Marina Venturini7Stefania Mitola8Piergiacomo Calzavara-Pinton9Dermatology Department, University of Brescia, ASST Spedali Civili di Brescia, 25123 Brescia, ItalyDermatology Department, University of Brescia, ASST Spedali Civili di Brescia, 25123 Brescia, ItalyDermatology Department, University of Brescia, ASST Spedali Civili di Brescia, 25123 Brescia, ItalyDermatology Department, University of Brescia, ASST Spedali Civili di Brescia, 25123 Brescia, ItalyDermatology Department, University of Brescia, ASST Spedali Civili di Brescia, 25123 Brescia, ItalyDepartment of Molecular and Translational Medicine, University of Brescia, 25123 Brescia, ItalyDermatology Department, University of Brescia, ASST Spedali Civili di Brescia, 25123 Brescia, ItalyDermatology Department, University of Brescia, ASST Spedali Civili di Brescia, 25123 Brescia, ItalyDepartment of Molecular and Translational Medicine, University of Brescia, 25123 Brescia, ItalyDermatology Department, University of Brescia, ASST Spedali Civili di Brescia, 25123 Brescia, ItalyIntroduction: 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. Materials and Methods: 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. Results: 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. Discussion: The SAAID values indicated a substantial impact of photoaging, with lower scores in photo-exposed elderly skin compared to non-exposed skin. Conclusions: 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.https://www.mdpi.com/2079-9284/12/3/111two-photon microscopyphotoagingextracellular matrixcollagen degradationaging skin
spellingShingle Stefano Bighetti
Chiara Rovati
Luca Bettolini
Mariachiara Arisi
Mariateresa Rossi
Cosetta Ravelli
Sara Rovaris
Marina Venturini
Stefania Mitola
Piergiacomo Calzavara-Pinton
Two-Photon Microscopy for the Investigation of Morphological and Quantitative Changes in Skin Chrono- and Photo-Aging
Cosmetics
two-photon microscopy
photoaging
extracellular matrix
collagen degradation
aging skin
title Two-Photon Microscopy for the Investigation of Morphological and Quantitative Changes in Skin Chrono- and Photo-Aging
title_full Two-Photon Microscopy for the Investigation of Morphological and Quantitative Changes in Skin Chrono- and Photo-Aging
title_fullStr Two-Photon Microscopy for the Investigation of Morphological and Quantitative Changes in Skin Chrono- and Photo-Aging
title_full_unstemmed Two-Photon Microscopy for the Investigation of Morphological and Quantitative Changes in Skin Chrono- and Photo-Aging
title_short Two-Photon Microscopy for the Investigation of Morphological and Quantitative Changes in Skin Chrono- and Photo-Aging
title_sort two photon microscopy for the investigation of morphological and quantitative changes in skin chrono and photo aging
topic two-photon microscopy
photoaging
extracellular matrix
collagen degradation
aging skin
url https://www.mdpi.com/2079-9284/12/3/111
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