Tissue engineering strategies to bioengineer the ageing skin phenotype in vitro

Human skin ageing is a complex and heterogeneous process, which is influenced by genetically determined intrinsic factors and accelerated by cumulative exposure to extrinsic stressors. In the current world ageing demographic, there is a requirement for a bioengineered ageing skin model, to further the understanding of the intricate molecular mechanisms of skin ageing, and provide a distinct and biologically relevant platform for testing actives and formulations. There have been many recent advances in the development of skin models that recapitulate aspects of the ageing phenotype in vitro. This review encompasses the features of skin ageing, the molecular mechanisms that drive the ageing phenotype, and tissue engineering strategies that have been utilised to bioengineer ageing skin in vitro

Kazrin, a novel periplakin-interacting protein associated with desmosomes and the keratinocyte plasma membrane

Periplakin forms part of the scaffold onto which the epidermal cornified envelope is assembled. The NH2-terminal 133 amino acids mediate association with the plasma membrane and bind a novel protein, kazrin. Kazrin is highly conserved and lacks homology to any known protein. There are four alternatively spliced transcripts, encoding three proteins with different NH2 termini. Kazrin is expressed in all layers of stratified squamous epithelia; it becomes membrane associated in the suprabasal layers, coincident with up-regulation of periplakin, and is incorporated into the cornified envelope of cultured keratinocytes. Kazrin colocalizes with periplakin and desmoplakin at desmosomes and with periplakin at the interdesmosomal plasma membrane, but its subcellular distribution is independent of periplakin. On transfection, all three kazrin isoforms have similar subcellular distributions. We conclude that kazrin is a novel component of desmosomes that associates with periplakin

Assessing the impact of airborne particulate pollution on human skin utilizing a novel human skin equivalent containing MUTZ ‐3‐derived Langerhans cells

Air pollution is an exogenous stressor known to have a detrimental impact on skin health through the induction of inflammation; however, the direct effect of topical pollution exposure is still being elucidated. Human skin equivalents (HSE) aim to reproduce in vitro the structure and function of the native skin tissue. However, HSEs typically lack skin‐resident immune cells, which could play a key role in the inflammatory response induced by pollution exposure. We outline the development of a HSE‐containing MUTZ‐3‐derived Langerhans cells (MUTZ‐3‐LCs), which show dendritic morphology and Langerhans cell marker expression. We demonstrated that HSE‐containing MUTZ‐3‐LC have lower basal levels of proinflammatory cytokines, but topical stimulation with allergens and irritant compounds induced a greater inflammatory response in these models compared to HSE without immune cells. To study the effect of pollution, we created a technique to apply diesel particulate matter (DPM) to HSEs. Though our microscopic analysis demonstrated that DPM does not penetrate the stratum corneum, we showed that DPM did induce production of proinflammatory cytokines, but notably only in HSEs containing MUTZ‐3‐LCs. These data suggest that topical exposure to air pollution can induce cutaneous inflammation and that skin‐resident immune cells contribute to this response. This highlights the significance of immune‐competent HSEs to the study of exogenous stressors in vitro

Development of a novel in vitro strategy to understand the impact of shaving on skin health: combining tape strip exfoliation and human skin equivalent technology

Introduction: The removal of unwanted hair is a widespread grooming practice adopted by both males and females. Although many depilatory techniques are now available, shaving remains the most common, despite its propensity to irritate skin. Current techniques to investigate the impact of shaving regimes on skin health rely on costly and lengthy clinical trials, which hinge on recruitment of human volunteers and can require invasive biopsies to elucidate cellular and molecular-level changes. Methods: Well-characterised human skin equivalent technology was combined with a commonplace dermatological technique of tape stripping, to remove cellular material from the uppermost layer of the skin (stratum corneum). This method of exfoliation recapitulated aspects of razor-based shaving in vitro, offering a robust and standardised in vitro method to study inflammatory processes such as those invoked by grooming practices. Results: Tape strip insult induced inflammatory changes in the skin equivalent such as: increased epidermal proliferation, epidermal thickening, increased cytokine production and impaired barrier function. These changes paralleled effects seen with a single dry razor pass, correlated with the number of tape strips removed, and were attenuated by pre-application of shaving foam, or post-application of moisturisation. Discussion: Tape strip removal is a common dermatological technique, in this study we demonstrate a novel application of tape stripping, to mimic barrier damage and inflammation associated with a dry shave. We validate this method, comparing it to razor-based shaving in vitro and demonstrate the propensity of suitable shave- and skin-care formulations to mitigate damage. This provides a novel methodology to examine grooming associated damage and a platform for screening potential skin care formulations

Palmitoylation Regulates Epidermal Homeostasis and Hair Follicle Differentiation

Palmitoylation is a key post-translational modification mediated by a family of DHHC-containing palmitoyl acyl-transferases (PATs). Unlike other lipid modifications, palmitoylation is reversible and thus often regulates dynamic protein interactions. We find that the mouse hair loss mutant, depilated, (dep) is due to a single amino acid deletion in the PAT, Zdhhc21, resulting in protein mislocalization and loss of palmitoylation activity. We examined expression of Zdhhc21 protein in skin and find it restricted to specific hair lineages. Loss of Zdhhc21 function results in delayed hair shaft differentiation, at the site of expression of the gene, but also leads to hyperplasia of the interfollicular epidermis (IFE) and sebaceous glands, distant from the expression site. The specific delay in follicle differentiation is associated with attenuated anagen propagation and is reflected by decreased levels of Lef1, nuclear β-catenin, and Foxn1 in hair shaft progenitors. In the thickened basal compartment of mutant IFE, phospho-ERK and cell proliferation are increased, suggesting increased signaling through EGFR or integrin-related receptors, with a parallel reduction in expression of the key differentiation factor Gata3. We show that the Src-family kinase, Fyn, involved in keratinocyte differentiation, is a direct palmitoylation target of Zdhhc21 and is mislocalized in mutant follicles. This study is the first to demonstrate a key role for palmitoylation in regulating developmental signals in mammalian tissue homeostasis

Collagenase Expression in Transgenic Mouse Skin Causes Hyperkeratosis and Acanthosis and Increases Susceptibility to Tumorigenesis

In a series of transgenic mice, the human tissue collagenase gene was expressed in the suprabasal layer of the skin epidermis. Visually, the mice had dry and scaly skin which upon histological analysis revealed acanthosis, hyperkeratosis, and epidermal hyperplasia. At the ultrastructural level, intercellular granular materials were absent in the transgenic skin epidermis but contact was maintained through the intact desmosomes. Despite a diversity of underlying etiologies, similar morphological hyperproliferative changes in the epidermis are observed in the human skin diseases of lamellar ichthyosis, atopic dermatitis, and psoriasis. Subsequent experiments demonstrate that when the transgenic mouse skin was treated once with an initiator (7,12-dimethyl-benz[a]anthracene) and then twice weekly with a promoter (12-O-tetradecanoylphorbol-13-acetate), there was a marked increase in tumor incidence among transgenic mice compared with that among control littermates. These experiments demonstrate that by overexpressing the highly specific proteolytic enzyme collagenase, a cascade of events leading to profound morphological changes which augment the sensitivity of the skin towards carcinogenesis is initiated in the epidermis