Abstract

Ultraviolet-A (UVA) irradiation induces harmful effects on skin cells and accelerates skin aging through oxidative stress. In this study, the effects of a hydrogen-generating silica material named ULH-002 against UVA injuries in human cells and 3D skin equivalents were investigated. The oxygen radical absorption capacity (ORAC) assay showed that both freshly prepared ULH-002 solutions and 7-day-old solutions exhibited equal peroxyl radical (ROO·) scavenging activities concentration-dependently. CellROX® green/orange staining showed that ULH-002 could reduce UVA-induced oxidative stress in human keratinocytes HaCaT and human gingival fibroblasts (HGFs). ULH-002 significantly prevented UVA-induced apoptotic/necrotic cell death and cell-viability decline in HGFs and keratinocytes, as shown by Annexin V/PI apoptosis assay and PrestoBlue assay, respectively. Immunostaining showed that ULH-002 prevented the UVA-induced deterioration of expression of both type IV and I collagens in the 3D skin equivalents, and similarly in monolayer HGFs. UVA-enhanced melanogenesis was observed in human melanocytes HMV-II and HMV-II cell-containing 3D skin equivalents, but markedly prevented by ULH-002 as demonstrated by Fontana–Masson’s staining. In conclusion, our data suggested that ULH-002 could protect human keratinocytes and fibroblasts from UVA-induced injuries, prevent the loss of type IV and I collagens, as well as reduce melanogenesis. ULH-002 might be developed as a skin care reagent in the cosmetic industry.

Highlights

  • Oxidative stress is the main contributor to skin aging processes

  • Keratinocytes, fibroblasts and melanocytes are the main cell-types in human skin

  • Oxidative stress-induced damage in these cells leads to cell number drop, collagen synthesis decline and pigmentation [7,8]

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Summary

Introduction

Oxidative stress is the main contributor to skin aging processes. Keratinocytes, fibroblasts and melanocytes are the main cell-types in human skin. Oxidative stress-induced damage in these cells leads to cell number drop, collagen synthesis decline and pigmentation [7,8]. Molecular hydrogen (H2 ) has gained attention as a selective antioxidant against toxic ROS. Many studies have reported that H2 exhibits antioxidative stress effects, and has various anti-inflammatory and anti-allergic effects [9,10,11,12]. We tested the effects of hydrogen-generating material named ULH-002 containing hydrogen-occluded microporous silica on ultravioletA (UVA)-induced cellular ROS generation, cell death, collagen loss and melanogenesis in human keratinocytes, fibroblasts, melanocytes and three-dimensional skin equivalents

Preparation of ULH-002 Solution
Cell Culture
Flow Cytometry
Fibrinogen Gel-Based Method
Collagen Gel-Based Method
UVA Irradiation
Cellular ROS Detection
Apoptosis Detection
2.10. Immunohistochemistry
2.11. Determination of Melanin Content
2.12. Statistical Analysis
Results
Characterization
EffectsUVA of ULH-002
Repressive of ULH-002 on UVA-induced cell damageininHGFs
Effects of on UVA-Induced
Repressive
Discussion
Conclusions
Full Text
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