In Vitro Study of Type I and III Collagen Expression by Active AAG1-AAI on With 2D Human Dermal Fibroblast Culture Using ELISA

Abstract

Aging is an essential and inevitable part of the biological process in living organisms. As a slow and chronic process, aging causes the gradual deterioration of physiological functions necessary for survival and fertility, such as deoxyribonucleic acid (DNA) repair and immune responses. The skin exhibits the most visible signs of aging due to its large volume and location on the body surface; creating an issue for most of the population which idealizes young and healthy skin. Hence, considerable efforts have been exerted in cosmetic and pharmaceutical sectors as a way to find remedies to delay or reverse aging (Kazanci, Kurus, & Atasever, 2016). Most anti-aging products target extracellular matrix (ECM) component production by fibroblasts, as its reduced production is one of the main mechanisms of dermal atrophy, a condition tightly related to aging. One of said ECM components, type I collagen, is an anti-aging marker as it constitutes the majority of collagens in the skin. Type I collagen typically constitutes 80% of the total collagen content in young skin, this is followed by type III collagen which constitutes 12%. With its role in providing tensile strength and elasticity to the skin, changes in collagen content would have visible results; as observed in skin wrinkling. It has been observed that the ratio between type I and type III collagen decreases with age, further emphasizing the significance of both collagen types in aging (Reilly & Lozano, 2021). With such findings, the anti-aging efficacy of a product can be measured from its ability to induce collagen production. To test cosmetic products, it is essential to have appropriate test systems to properly simulate the complexity of human skin; a property which is not represented in two-dimensional (2D) cell cultures. Hence, artificial skin equivalents such as the EpiDerm™ Skin Model can be used as a solution. The ability of skin equivalents to simulate processes involved in the skin, such as penetration, makes it ideal for usage in this study (Neupane et al., 2020). This study aims to evaluate the effect of active AAG1-AAI on type I and III collagen in human dermal fibroblasts (HDF) with the EpiDermTM Skin Model artificial skin equivalent as a skin barrier model by using enzyme-linked immunosorbent assay (ELISA).