Characterization of a New Reconstructed Full Thickness Skin Model, T-Skin™, and its Application for Investigations of Anti-Aging Compounds

Abstract

Background: We have characterized a new reconstructed full-thickness skin model, T-Skin™, compared to normal human skin (NHS) and evaluated its use in testing anti-aging compounds. Methods: The structure and layer-specific markers were compared with NHS using histological and immunohistological staining. In anti-aging experiments, T-SkinTM was exposed to retinol (10 µM) or vitamin C (200 µM) for 5 days, followed by immunohistological staining evaluation. Results: T-Skin™ exhibits a well stratified, differentiated and self-renewing epidermis with a dermal compartment of functional fibroblasts. Epidermal (cytokeratin 10, transglutaminase 1), dermo–epidermal junction (DEJ) (laminin 5, collagen-IV, collagen VII) and dermally-located (fibrillin 1, procollagen I) biomarkers were similar to those in NHS. Treatment of T-Skin™ with retinol decreased the expression of differentiation markers, cytokeratin 10 and transglutaminase 1 and increased the proliferation marker, Ki67, in epidermis basal-layer cells. Vitamin C increased the expression of DEJ components, collagen IV and VII and dermal procollagen 1. Conclusions: T-Skin™ exhibits structural and biomarker location characteristics similar to NHS. Responses of T-Skin™ to retinol and vitamin C treatment were consistent with those of their known anti-aging effects. T-Skin™ is a promising model to investigate responses of epidermal, DEJ and dermal regions to new skin anti-ageing compounds.

Topical application of a commercially available formulation of vitamin C stabilized by vitamin E and ferulic acid reduces tissue viability and protein synthesis in ex vivo human normal skin

“…Vitamin C is a powerful free radical scavenger observed in high concentrations of the epidermis and dermis, protecting skin against ROS‐induced oxidative damage 9,14 . However, UV‐induced damage may reduce the antioxidant defense system in skin, including the levels of vitamin C 5 .…”

“…Recently, it was showed that vitamin C encapsulated into liposomes enhances the deposition of the collagen type I in mouse embryo fibroblasts after 48 hours 38 . In artificial human skin, the administration of vitamin C promotes the synthesis of the collagen types VII and IV, laminin V, and procollagen type I in the dermis after 5 days 14 . In our study, formulation 1 diluted at 50% reduced the gene expression of ECM components (Col1a1, Fbn1, and Eln), protein and gene levels of housekeeping markers (actin, Rlp10, and GAPDH), and cell proliferation (lower PCNA levels) in the human skin ex vivo.…”

“…In the epidermis, vitamin C promotes keratinocyte differentiation and epidermal barrier formation, stimulating the synthesis of ceramides 7‐9 . In the dermis, vitamin C promotes the synthesis of elastic and collagen fibers in humans and rodents 10‐14 . In addition, topical administration of vitamin C protects skin against UV radiation–induced oxidative damage in pig models or reconstituted human epidermis 15‐17 .…”

A Quantitative MALDI-MSI Study of the Movement of Molecules in Biological Systems

“…Over the years tissue engineered HSEs have been established as valid models for in-vitro cosmetic and pharmaceutical testing, as well as for the investigation of skin biology mechanisms behind the generation of the epidermis, skin barrier repair/wound healing, skin pathologies and absorption testing (Schäfer-Korting, Mahmoud, et al, 2008;Xie et al, 2010;Ali et al, 2015;De Vuyst et al, 2017;Lewis et al, 2018;Bataillon et al, 2019).…”

“…Chapter 7: Conclusion and future work Conclusion 3D in-vitro tissue models of human skin represent a valid alternative to monolayer 2D cell culture, ex-vivo human and animal skin models, and, at the present time, their application finds a place in many skin research fields (Schäfer-Korting, Mahmoud, et al, 2008;Xie et al, 2010;Ali et al, 2015;De Vuyst et al, 2017;Lewis et al, 2018;Bataillon et al, 2019). 3D in-vitro skin models offer several advantages; they have a higher resemblance to the in-vivo human skin microenvironment compared to monolayer 2D cell culture, they guarantee a higher quality of preservation compared to ex-vivo skin, as they are still living systems they are easy to obtain without requiring an individual ethical licence, and they represent a valid replacement to animal testing in line with the principle of the UK organisation 3Rs (Replacement, Reduction and Refinement).…”