Cosmetic Testing Research Articles

Full-Thickness Perfused Skin-on-a-Chip with In Vivo-Like Drug Response for Drug and Cosmetics Testing

In this study, we present a novel 3D perfused skin-on-a-chip model fabricated using micro-precision 3D printing, which offers a streamlined and reproducible approach for incorporating perfusion. Perfused skin models are well-regarded for their advantages, such as improved nutrient supply, enhanced barrier function, and prolonged tissue viability. However, current models often require complex setups, such as self-assembled endothelial cells or sacrificial rods, which are prone to variability and time-consuming. Our model uses projection micro-stereolithography 3D printing to create precise microcapillary-like channels using a biocompatible resin, overcoming the drug-absorbing properties of PDMS. A customized chip holder allows for the simultaneous culture of six perfused chips, enabling high-throughput testing. The engineered skin-on-a-chip features distinct dermis and epidermis layers, confirmed via H&E staining and immunostaining. To evaluate drug screening capabilities, inflammation was induced using TNF-α and treated with dexamethasone, with cytokine levels compared to 2D cultures and human skin biopsies. Our 3D model exhibited drug response trends similar to human skin, while showing reduced cytotoxicity over time compared to biopsies. This perfused skin-on-a-chip provides a reliable, physiologically relevant alternative for drug and cosmetics screening, simplifying perfusion setup while preserving key benefits.

Historical Analysis of Cosmetics Testing Development and Alternatives

Historical Analysis of Cosmetics Testing Development and Alternatives

Guidelines for Manufacturing and Application of Organoids: Skin.

To address the limitations of animal testing, scientific research is increasingly focused on developing alternative testing methods. These alternative tests utilize cells or tissues derived from animals or humans for in vitro testing, as well as artificial tissues and organoids. In western countries, animal testing for cosmetics has been banned, leading to the adoption of artificial skin for toxicity evaluation, such as skin corrosion and irritation assessments. Standard guidelines for skin organoid technology becomes necessary to ensure consistent data and evaluation in replacing animal testing with in vitro methods. These guidelines encompass aspects such as cell sourcing, culture techniques, quality requirements and assessment, storage and preservation, and organoid-based assays.

Computational Methods as Part of Scientific Research in Cosmetic Sciences—Are We Using the Opportunity?

In the field of the cosmetic industry, significant efforts are made to develop methods that are not only cost effective and time effective but are also environmentally friendly and cruelty free. Cosmetic tests using in vivo animal models are currently banned in the European Union. To fulfil regulatory requirements, new approach methodologies (NAMs) are implemented, and thereupon, in silico techniques have constantly acquired significance. This review aimed to show the general picture of the available computational methods and approaches, give some examples of their applications, present capabilities and limitations, and propose the way forward. The general information about in silico modelling and examples of its usage in the context of cosmetics and its legal regulation are presented. The review is divided with a focus on three endpoints of interest: (1) safety assessment, (2) exposure assessment, and (3) formulation characterization. With this comprehensive analysis, we try to answer the question as to whether we are using the opportunity.

Moving lab-grown tissues into the clinic: organ-on-a-chip and bioengineered skin systems

For patients with end stage organ failure, organ transplant is frequently the only curative option available. However, organs available for transplant are in critically short supply around the world, which has led to lengthy wait times and increased mortality. Increased global life expectancy, coupled with raised age thresholds for recipients, has heightened demand and further compounded the need for alternative strategies. Bioengineering substitutes including organ-on-a-chip and 3D bioprinting technologies have made considerable strides toward whole organ generation. Skin is the organ where the most advances have been made thus far, due to the relatively less complex spatial architecture and industry interest in the development of sophisticated models for pharmaceutical and cosmetics testing. Here, we discuss the challenges of recapitulating the complexity of native skin, including a stratified structure, vascularization, and inclusion of skin appendages, such as hair follicles and sweat glands. We discuss current technological and biological progress in the field of tissue and organ bioengineering as well as highlight future challenges to generate de novo tissue for skin grafting.

Review: Policy strategy of nano cosmetic testing in Indonesia

The implementation of nanotechnology in the cosmetic industry has developed promptly in recent decades. Among them are sunscreen, skincare, decorative makeup, and hair colorant. The cosmetics industry is a significant user of nanoscale raw materials in its manufacturing procedures. However, the supervision regarding the safety and quality of cosmetic products containing nanomaterials (NMs) in Indonesia conducted by the Indonesian FDA remains not fully optimized. This poses a challenge for the Indonesian FDA, particularly concerning the significance of employing appropriate characterization methods and the necessity for systematic evaluation. Hazard identification, exposure evaluation, microbiological testing, and NM characterization are some methods that can be conducted for monitoring nano cosmetics. Procuring instruments for nano-analysis will be a significant investment for the purpose of monitoring cosmetic products marketed in Indonesia.

Culture insert device with perfusable microchannels enhances in vitro skin model development and barrier function assessment

The ever-stricter regulations on animal experiments in the field of cosmetic testing have prompted a surge in skin-related research with a special focus on recapitulation of the in vivo skin structure in vitro. In vitro human skin models are seen as an important tool for skin research, which in recent years attracted a lot of attention and effort, with researchers moving from the simplest 2-layered models (dermis with epidermis) to models that incorporate other vital skin structures such as hypodermis, vascular structures, and skin appendages. In this study, we designed a microfluidic device with a reverse flange-shaped anchor that allows culturing of an in vitro skin model in a conventional 6-well plate and assessing its barrier function without transferring the skin model to another device or using additional contraptions. Perfusion of the skin model through vascular-like channels improved the morphogenesis of the epidermis compared with skin models cultured under static conditions. This also allowed us to assess the percutaneous penetration of the tested caffeine permeation and vascular absorption, which is one of the key metrics for systemic drug exposure evaluation.

Assessment of the three-test genetic toxicology battery for groundwater metabolites.

The two-test in vitro battery for genotoxicity testing (Ames and micronucleus) has in the majority of cases replaced the three-test battery (as two-test plus mammalian cell gene mutation assay) for the routine testing of chemicals, pharmaceuticals, cosmetics, and agrochemical metabolites originating from food and feed as well as from water treatment. The guidance for testing agrochemical groundwater metabolites, however, still relies on the three-test battery. Data collated in this study from 18 plant protection and related materials highlights the disparity between the often negative Ames and in vitro chromosome aberration data and frequently positive in vitro mammalian cell gene mutation assays. Sixteen of the 18 collated materials with complete datasets were Ames negative, and overall had negative outcomes in in vitro chromosome damage tests (weight of evidence from multiple tests). Mammalian cell gene mutation assays (HPRT and/or mouse lymphoma assay (MLA)) were positive in at least one test for every material with this data. Where both MLA and HPRT tests were performed on the same material, the HPRT seemed to give fewer positive responses. In vivo follow-up tests included combinations of comet assays, unscheduled DNA synthesis, and transgenic rodent gene mutation assays, all gave negative outcomes. The inclusion of mammalian cell gene mutation assays in a three-test battery for groundwater metabolites is therefore not justified and leads to unnecessary in vivo follow-up testing.

Applications of Engineered Skin Tissue for Cosmetic Component and Toxicology Detection.

The scale of the cosmetic market is increasing every day. There are many safety risks to cosmetics, but they benefit people at the same time. The skin can become red, swollen, itchy, chronically toxic, and senescent due to the misuse of cosmetics, triggering skin injuries, with contact dermatitis being the most common. Therefore, there is an urgent need for a system that can scientifically and rationally detect the composition and perform a toxicological assessment of cosmetic products. Traditional detection methods rely on instrumentation and method selection, which are less sensitive and more complex to perform. Engineered skin tissue has emerged with the advent of tissue engineering technology as an emerging bioengineering technology. The ideal engineered skin tissue is the basis for building good in vitro structures and physiological functions in this field. This review introduces the existing cosmetic testing and toxicological evaluation methods, the current development status, and the types and characteristics of engineered skin tissue. The application of engineered skin tissue in the field of cosmetic composition detection and toxicological evaluation, as well as the different types of tissue engineering scaffold materials and three-dimensional (3D) organoid preparation approaches, is highlighted in this review to provide methods and ideas for constructing the next engineered skin tissue for cosmetic raw material component analysis and toxicological evaluation.

Chemical Profile and Health-Promoting Activities of Crataegus laciniata (Rosaceae) Flowers.

In the present study, we focused our attention on Crataegus laciniata Ucria (Rosaceae), which is wild growing in western Sicily (Italy). The chemical profile of the C. laciniata flower's (CLF) ethanolic (70%) extract showed the presence of both C-flavonoid and O-flavonoid derivatives. Beyond the main metabolites, like hyperoside and vitexin, there are several luteolin derivates, in addition to catechin and epicatechin dimers or trimers. Regarding the antioxidant activities, CLF showed a strong ability to scavenge DPPH and ABTS radicals and a good Fe3+-reducing antioxidant power. The investigation into the key enzymes in diabetes showed strong inhibition on α-amylase and α-glucosidase, whereas the skin-whitening properties are linked to inhibitory effects on tyrosinase. Moreover, we employed Danio rerio (zebrafish) for toxicity assessment, as it represents an ideal in vivo model due to its high correlation with humans in response to pharmaceutical and cosmetic testing. Zebrafish embryos exposed to CLF (25-100 µg/mL) showed marked depigmentation compared to phenylthiourea (PTU), in addition to a high survival percentage and the absence of malformations. In conclusion, this experimental study outlines that C. laciniata flowers could be a potential source of bioactive compounds for application in the pharmaceutical and cosmeceutical industries.

Biomimetic human skin model patterned with rete ridges

Rete ridges consist of undulations between the epidermis and dermis that enhance the mechanical properties and biological function of human skin. However, most human skin models are fabricated with a flat interface between the epidermal and dermal layers. Here, we report a micro-stamping method for producing human skin models patterned with rete ridges of controlled geometry. To mitigate keratinocyte-induced matrix degradation, telocollagen–fibrin matrices with and without crosslinks enable these micropatterned features to persist during longitudinal culture. Our human skin model exhibits an epidermis that includes the following markers: cytokeratin 14, p63, and Ki67 in the basal layer, cytokeratin 10 in the suprabasal layer, and laminin and collagen IV in the basement membrane. We demonstrated that two keratinocyte cell lines, one from a neonatal donor and another from an adult diabetic donor, are compatible with this model. We tested this model using an irritation test and showed that the epidermis prevents rapid penetration of sodium dodecyl sulfate. Gene expression analysis revealed differences in keratinocytes obtained from the two donors as well as between 2D (control) and 3D culture conditions. Our human skin model may find potential application for drug and cosmetic testing, disease and wound healing modeling, and aging studies.

LB1669 Development and characterization of a miniaturized in-vitro skin model on a chip for high-throughput cosmetic testing

LB1669 Development and characterization of a miniaturized in-vitro skin model on a chip for high-throughput cosmetic testing

LB1668 Anti-aging effects of autophagy analyzed within a 3D microfluidic skin-on-a-chip for skin cosmetics testing

LB1668 Anti-aging effects of autophagy analyzed within a 3D microfluidic skin-on-a-chip for skin cosmetics testing

Analysis of facial redness by comparing VISIA and YLGTD.

Erythema, characterized by redness of the skin, is a common symptom in various facial skin conditions. Recent advancements in image processing and analysis techniques have led to the development of methods for analyzing and assessing skin texture. This study aimed to investigate the correlation between the parameters of "You Look Good Today" (YLGTD) and VISIA in the detection and assessment of facial redness. Thirty female subjects participated in this experiment, undergoing assessments using both YLGTD and VISIA. The subjects were evaluated for facial redness, and the feature count results within the red zone were measured by VISIA. YLGTD analyzed the number and percentage of red zone pixels. The assessments were conducted between [specific dates] in [location]. The results demonstrated a significant positive correlation between the feature count results within the red zone measured by VISIA and the number of red zone pixels. Similarly, YLGTD exhibited a significant positive correlation with the number and percentage of red zone pixels. In conclusion, our findings suggest a correlation between YLGTD and VISIA in the measurement of facial erythema. YLGTD can serve as a portable device for primary screening assessments, offering a convenient and reliable method to evaluate facial redness. This research contributes to the development of non-invasive techniques for assessing and monitoring facial skin conditions, providing valuable insights for dermatological diagnosis and cosmetic testing.

Development of a lip vermilion epithelium reconstruction model using keratinocytes from skin and oral mucosa.

Lip vermilion is unique and can be distinguished from the adjacent skin and oral mucosa. However, because of the lack of appropriate evaluation tools, skin and/or oral mucosa substitutes such as in vitro vermilion epithelial models have been used for lip product testing. We aimed to develop and characterize a lip vermilion epithelium reconstruction model (LVERM) using skin and oral keratinocytes. LVERM was manufactured by co-culturing primary skin and oral keratinocytes, using a device that allowed the separation of cell seeding, and created an intercalated cell-free zone, referred to as the vermilion part. After removing the device, LVERM construction was completed in 8 days, in a submerged condition. Subsequently, they were placed in an air-liquid interface for 7 days. To determine the epithelial characteristics of LVERM, keratin 2e (KRT2) and small proline-rich protein 3 (SPRR3) expression patterns were examined. The in vivo expression profiles of KRT2 and SPRR3 genes in vermilion were also examined. We found that a continuous multi-layered epithelium was generated in the LVERM that exhibited ortho- and para-keratinization in the skin and oral mucosa parts, respectively. Although an intermediate keratinization pattern was observed in the vermilion part, KRT2 and SPRR3 were co-expressed in the suprabasal layer, consistent with the expression pattern of a single vermilion epithelial model. Clustering analysis revealed that KRT2 and SPRR3 gene expression in vermilion was location-dependent within the sample. Therefore, LVERM can be used as an evaluation tool for lip products and has great importance in innovative approaches for cosmetic testing.

Save the Trip to the Store: Sustainable Shopping, Electronic Word of Mouth on Instagram and the Impact on Cosmetic Purchase Intentions

The COVID-19 pandemic has forced many brands to stop using cosmetic testers to avoid the risk of spreading the infection, jeopardising the future of cosmetic testing. Consequently, consumers must find alternative methods to conduct their information searches and, more importantly, the prospects of shopping online without going to the store to test the product. With the enormous prospects of social media cosmetic electronic word of mouth (eWOM), it is imperative to examine the influence of cosmetic eWOM on social media and for cosmetic marketers to understand the antecedents that result in cosmetic consumers making a purchase. The adapted information adoption model was validated through structural equation modelling based on 341 eligible surveys. The results confirmed that information quality, source credibility, information usefulness, and information adoption are the key antecedents in eWOM on Instagram when investigating purchase intentions in the colour cosmetic industry. This study is one of the pioneers in empirically testing the relationship between information quality and source credibility on information usefulness and, subsequently, the relationship between information usefulness, information adoption, and purchase intentions in a western market based on the cosmetic industry. These new insights provide practical implications for a cosmetic marketer, suggesting the key variables leading to purchase intentions in cosmetic eWOM, which can be utilised in marketing techniques.

Comparison of structural characteristics and molecular markers of rabbit skin, pig skin, and reconstructed human epidermis for an ex vivo human skin model

The Organization for Economic Co-operation and Development approved a reconstructed human epidermis (RHE) model for in vitro skin irritation and corrosion tests as an alternative to animal testing for cosmetics, which has been banned in the European Union since 2013. However, RHE models have several limitations, such as high manufacturing costs, a loose skin barrier, and inability to simulate all cellular and non-cellular components of the human epidermis. Therefore, new alternative skin models are needed. Ex vivo skin models have been suggested as promising tools. Here, we investigated the structural similarities in the epidermis of pig and rabbit skin, a commercial RHE model (Keraskin), and human skin. To compare the structural similarity, the thickness of each epidermal layer was compared using molecular markers. Among the candidate human skin surrogates, the epidermal thickness of the pig skin was the most similar to that of human skin, followed by rabbit skin and Keraskin. Keraskin showed thicker cornified and granular layers than human skin, while rabbit skin displayed thinner layers. Moreover, the proliferation indices of Keraskin and rabbit skin were higher than those of human skin, whereas the proliferation index of the pig skin was similar to that of human skin. Some or none of the human skin barrier proteins FLG, CLDN1, and CDH1 were expressed in pig and rabbit skin, whereas all human proteins were expressed in Keraskin. Collectively, we propose ex vivo pig skin as the most suitable model for skin irritation testing because of its similarity to human skin.

The 19th FRAME Annual Lecture, November 2022: Safer Chemicals and Sustainable Innovation Will Be Achieved by Regulatory Use of Modern Safety Science, Not by More Animal Testing.

The decisions we make on chemical safety, for consumers, workers and the environment, must be based on the best scientific data and knowledge available. Rapid advances in biology, in cell-based technologies and assays, and in analytical and computational approaches, have led to new types of highly relevant scientific data being generated. Such data enable us to improve the safety decisions we make, whilst also enabling us to avoid animal testing. Stimulated by the UK and EU bans on animal testing for cosmetics, Next Generation Risk Assessment (NGRA) approaches, which integrate various types of non-animal scientific data, have been established for assessing the safety of chemical ingredients used in cosmetics and other consumer products. In stark contrast, the chemicals regulations in Europe and other parts of the world have not kept pace with modern safety science and regulators are now mandating even more animal testing. Urgently closing this science-regulation gap is essential to upholding the EU's legislative requirement that any animal testing is a last resort. The ongoing revisions of UK and EU chemicals strategy and regulations provide an opportunity to fundamentally change the design and assessment paradigm needed to underpin safe and more sustainable innovation, through applying the best science and tools available rather than continuing to be anchored in animal tests dating back many decades. A range of initiatives have recently been launched in response to this urgent need, in the UK as well as in the EU.

Cosmetic Testing Equipment: Device and Types of Equipment for Dermatological Evaluation for Women’s Skin

There are many different equipment and techniques for an aesthetic skin evaluation, efficacy testing, claim support, and objective measurements of women’s skin parameters for all applications. Women have softer skin than men. Nothing commercially available in terms of moisture, oiliness, color, texture, etc existed over 20 years ago. Fortunately, several cost-effective ways are available to quantify these characteristics and compare them before and after a treatment or application. Before distributing the finished product to outside testing facilities for the final validation, the cosmetic business conducts these tests at different stages of product development to determine the product’s direction. In this review, we have summarized the C+K devices that are acknowledged as industry-standard tools for assessing efficacy in the fields of cosmetics and the types of equipment used for the cosmetic evaluation of the skin.

CEpiderm, a Canine Skin Analog Suitable for In Vivo Testing Replacement

Skin is one of the organs most tested for toxicity and safety evaluation during the process of drug research and development and in the past has usually been performed in vivo using animals. Over the last few years, non-animal alternatives have been developed and validated epidermis models for human and rat skin are already available. Our goal was to develop a histotypical canine skin analog, suitable for non-animal biocompatibility and biosafety assessment. Canine keratinocytes were seeded in an air-lift culture using an adapted version of the CELLnTEC protocol. Corrosion and irritation protocols were adapted from human EpiSkinTM. For histological analysis, sample biopsies were fixed in neutral-buffered formalin, and paraffin slices were routinely processed and stained with hematoxylin and eosin. A canine multilayer and stratified epidermal-like tissue (cEpiderm), confirmed by histological analysis, was obtained. The cEpiderm tissue exhibited normal morphological and functional characteristics of epidermis, namely impermeability and an adequate response to stressors. The cEpiderm is a promising canine skin model for non-animal safety testing of veterinary pharmaceuticals and/or cosmetics, significantly contributing to reducing undesirable in vivo approaches. cEpiderm is therefore a valid canine skin model and may be made commercially available either as a service or as a product.