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      Application of ultrasound-mediated adapalene-coated lysozyme-shelled microbubbles in UVA-induced skin photoaging

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          Abstract

          Photoaging, the premature aging of skin induced by ultraviolet rays, is characterized by wrinkling, roughness, laxity, and pigmentary changes. Various natural and synthetic retinoids have been explored for the treatment of aging. Among retinoids, adapalene (Ada, 0.3%) is one of the most potent and widely used drugs to treat photoaging. However, it causes irritant reactions that limit its acceptance by patients. Several studies have shown the applicability of Lysozyme (Lys)-shelled microbubbles (MBs) for drug delivery through sonophoresis, and recently we have shown its efficiency to treat inflammatory skin disease. Here, we report the construction of novel Ada-LysMBs based on opposite electric charges for combined effects to treat photoaging. The Ada-LysMBs were self-assembled and had a mean diameter of 2857 nm. The maximum loading efficiency of Ada onto LysMBs was 13.99 ± 0.59%. An acoustic power density of 3 W/cm 2 for 1 min revealing maximum penetration depth of LysMBs was optimized for further in vitro and in vivo studies of Ada-LysMBs. It was observed that in vitro Ada release from Ada-LysMBs at 6 h after ultrasound (US) treatment was more rapid at pH 7.4 (82%) than at pH 5.5 (73%). Franz diffusion experiments on isolated porcine skin indicated that US approximately doubled Ada delivery by Ada-LysMBs and Ada + LysMBs at 12 h and six-fold Lys permeation by LysMBs at 6 h, compared to these treatments alone. A 5-week in vivo study in mice identified significant wrinkle reduction in animals treated with US plus Ada-LysMBs. Our findings indicate that US may be used with Ada-LysMBs in the water phase to treat photoaging by normalizing hyperkeratinization and promoting collagen synthesis.

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          Photoageing: mechanism, prevention and therapy.

          Photoageing is the superposition of chronic ultraviolet (UV)-induced damage on intrinsic ageing and accounts for most age-associated changes in skin appearance. It is triggered by receptor-initiated signalling, mitochondrial damage, protein oxidation and telomere-based DNA damage responses. Photodamaged skin displays variable epidermal thickness, dermal elastosis, decreased/fragmented collagen, increased matrix-degrading metalloproteinases, inflammatory infiltrates and vessel ectasia. The development of cosmetically pleasing sunscreens that protect against both UVA and UVB irradiation as well as products such as tretinoin that antagonize the UV signalling pathways leading to photoageing are major steps forward in preventing and reversing photoageing. Improved understanding of the skin's innate UV protective mechanisms has also given rise to several novel treatment concepts that promise to revolutionize this field within the coming decade. Such advances should not only allow for the improved appearance of skin in middle age and beyond, but also greatly reduce the accompanying burden of skin cancer.
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            Retinoids in the treatment of skin aging: an overview of clinical efficacy and safety

            Aging of skin is an intricate biological process consisting of two types. While intrinsic or chronological aging is an inevitable process, photoaging involves the premature aging of skin occurring due to cumulative exposure to ultraviolet radiation. Chronological and photoaging both have clinically differentiable manifestations. Various natural and synthetic retinoids have been explored for the treatment of aging and many of them have shown histological and clinical improvement, but most of the studies have been carried out in patients presenting with photoaged skin. Amongst the retinoids, tretinoin possibly is the most potent and certainly the most widely investigated retinoid for photoaging therapy. Although retinoids show promise in the treatment of skin aging, irritant reactions such as burning, scaling or dermatitis associated with retinoid therapy limit their acceptance by patients. This problem is more prominent with tretinoin and tazarotene whereas other retinoids mainly represented by retinaldehyde and retinol are considerably less irritating. In order to minimize these side effects, various novel drug delivery systems have been developed. In particular, nanoparticles have shown a good potential in improving the stability, tolerability and efficacy of retinoids like tretinoin and retinol. However, more elaborate clinical studies are required to confirm their advantage in the delivery of topical retinoids.
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              Solar UV radiation reduces the barrier function of human skin.

              The ubiquitous presence of solar UV radiation in human life is essential for vitamin D production but also leads to skin photoaging, damage, and malignancies. Photoaging and skin cancer have been extensively studied, but the effects of UV on the critical mechanical barrier function of the outermost layer of the epidermis, the stratum corneum (SC), are not understood. The SC is the first line of defense against environmental exposures like solar UV radiation, and its effects on UV targets within the SC and subsequent alterations in the mechanical properties and related barrier function are unclear. Alteration of the SC's mechanical properties can lead to severe macroscopic skin damage such as chapping and cracking and associated inflammation, infection, scarring, and abnormal desquamation. Here, we show that UV exposure has dramatic effects on cell cohesion and mechanical integrity that are related to its effects on the SC's intercellular components, including intercellular lipids and corneodesmosomes. We found that, although the keratin-controlled stiffness remained surprisingly constant with UV exposure, the intercellular strength, strain, and cohesion decreased markedly. We further show that solar UV radiation poses a double threat to skin by both increasing the biomechanical driving force for damage while simultaneously decreasing the skin's natural ability to resist, compromising the critical barrier function of the skin.
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                Author and article information

                Contributors
                Role: ConceptualizationRole: Funding acquisitionRole: InvestigationRole: MethodologyRole: Writing – original draft
                Role: Data curationRole: Formal analysisRole: Investigation
                Role: MethodologyRole: SupervisionRole: Writing – review & editing
                Role: Data curationRole: InvestigationRole: Software
                Role: Formal analysisRole: Funding acquisitionRole: InvestigationRole: MethodologyRole: Writing – review & editing
                Role: Data curationRole: Formal analysisRole: MethodologyRole: Writing – original draftRole: Writing – review & editing
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                21 May 2020
                2020
                : 15
                : 5
                : e0232617
                Affiliations
                [1 ] Graduate Institute of Biomedical Engineering, National Taiwan University of Science and Technology, Taipei, Taiwan
                [2 ] Department of Medical Engineering, National Defense Medical Center, Taipei, Taiwan
                [3 ] Graduate Institute of Manufacturing Technology, National Taipei University of Technology, Taipei, Taiwan
                [4 ] Department of Pathology, Tri-Service General Hospital, National Defense Medical Center, Taipei, Taiwan
                [5 ] Department of Dermatology, Tri-Service General Hospital, National Defense Medical Center, Taipei, Taiwan
                [6 ] Department of Biochemistry, National Defense Medical Center, Taipei, Taiwan
                Argonne National Laboratory, UNITED STATES
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                Author information
                http://orcid.org/0000-0001-9101-6662
                http://orcid.org/0000-0003-3075-2118
                Article
                PONE-D-19-32633
                10.1371/journal.pone.0232617
                7242023
                32438389
                49ebd313-1c3b-4bb9-accf-4fe2740b9919
                © 2020 Liao et al

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 26 November 2019
                : 18 April 2020
                Page count
                Figures: 10, Tables: 3, Pages: 21
                Funding
                Funded by: funder-id http://dx.doi.org/10.13039/501100004663, Ministry of Science and Technology, Taiwan;
                Award ID: MOST106-2221-E-011-043-MY3
                Award Recipient :
                Funded by: funder-id http://dx.doi.org/10.13039/501100010425, Tri-Service General Hospital;
                Award ID: TSGH-NTUST-106-02
                Award Recipient :
                Funded by: funder-id http://dx.doi.org/10.13039/501100010425, Tri-Service General Hospital;
                Award ID: TSGH-C106-062
                Award Recipient :
                Funded by: funder-id http://dx.doi.org/10.13039/501100010425, Tri-Service General Hospital;
                Award ID: TSGH-C107-061
                Award Recipient :
                Funded by: funder-id http://dx.doi.org/10.13039/501100010425, Tri-Service General Hospital;
                Award ID: TSGH-C108-034
                Award Recipient :
                Funded by: funder-id http://dx.doi.org/10.13039/501100010425, Tri-Service General Hospital;
                Award ID: MAB107-034
                Award Recipient :
                This work was supported in part by grants from the Ministry of Science and Technology, Taiwan (MOST106-2221-E-011-043-MY3 to AHL), the Tri-Service General Hospital, Taiwan (TSGH-NTUST-106-02 to AHL, TSGH-C106-062, TSGH-C107-061 to YCL and TSGH-C108-034 to CPC) and the Ministry of National `Defense, Taiwan (MAB107-034 to CPC).
                Categories
                Research Article
                Research and Analysis Methods
                Specimen Preparation and Treatment
                Mechanical Treatment of Specimens
                Sonication
                Biology and Life Sciences
                Biochemistry
                Enzymology
                Enzymes
                Lysozyme
                Biology and Life Sciences
                Biochemistry
                Proteins
                Enzymes
                Lysozyme
                Research and Analysis Methods
                Animal Studies
                Experimental Organism Systems
                Model Organisms
                Mouse Models
                Research and Analysis Methods
                Model Organisms
                Mouse Models
                Research and Analysis Methods
                Animal Studies
                Experimental Organism Systems
                Animal Models
                Mouse Models
                Biology and Life Sciences
                Biochemistry
                Proteins
                Collagens
                Biology and Life Sciences
                Anatomy
                Integumentary System
                Skin
                Epidermis
                Medicine and Health Sciences
                Anatomy
                Integumentary System
                Skin
                Epidermis
                Engineering and Technology
                Equipment
                Measurement Equipment
                Spectrometers
                Spectrophotometers
                Engineering and Technology
                Membrane Technology
                Membrane Dialysis
                Research and Analysis Methods
                Separation Processes
                Molecular Dialysis
                Membrane Dialysis
                Physical Sciences
                Physics
                Acoustics
                Custom metadata
                All relevant data are within the manuscript and its Supporting Information file.

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