TESTING
Novel 3D model to study feminine care ingredients
Thomas Clamens, Céline Lancelot, Michel Salmon – Straticell
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The human vaginal epithelium is a non- keratinized stratified squamous epithelium composed of basal, parabasal, intermediate, and superficial layers that undergo dynamic remodeling across the menstrual cycle but also with woman’s age and hormonal status.1 The basal layer is the deepest layer, which
is mitotically active, continuously producing new cells. The parabasal cells are small, round cells that emerge from the basal layer and form several layers of tissue. The intermediate cells are characterized by foamy nuclei and larger cytoplasm rich in glycogen. The most superficial layer consists of
flattened, non-cornified cells with pyknotic nuclei.2
This multilayered structure forms a
robust barrier that protects underlying tissues from mechanical stress and chemical insults. In a woman’s life, main external stressors
are hygiene soaps, moisturizing creams, local contraceptives or even menstrual pads. Yet, because those intimate care products are in direct and often prolonged contact with the vaginal tissue, ingredients with even sub-irritant effects can disrupt the epithelial barrier function and induce inflammation. This is why feminine care products are
subject to regulations aiming at protecting consumers from hazardous chemicals, including the assessment of the level of risk by in vitro safety assays ensuring the compatibility of topically applied women’s health products and devices with the vaginal tissues. The traditional method for safety testing has long been based on ex vivo animal vaginal
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tissues.3
The shift in recent years toward non-
animal and more ethical alternatives allowed the emergence of innovative in vitro models. Conventional vaginal epithelial cell lines
in monolayer culture are now available from providers. They are highly adapted to preliminary researches and early identifications of potential safety issues but like every 2D cell systems, fail to replicate the morphology and function of multilayered structure such as the vaginal epithelium. In contrast, cells can be cultured in specific
conditions to generate organ-like 3D systems that replicate the architecture and behaviour of native tissues. Such organotypic structures are more representative of the in vivo environment and considered nowadays as the gold standard in safety and efficacy assessment in preclinical studies.4,5
Notably, organoid-based assays
have demonstrated higher predictive accuracy for irritation and inflammation compared to monolayer cell cultures, providing a more reliable cell system for assessing product safety.6
The present study reports the newly
reconstruction of a vaginal organoid tissue model and demonstrates its use to evaluate the safety of topical ingredients commonly found in feminine care formulations. The in vitro 3D HVE model described here
was reconstructed from isolated A431 vulval epidermoid carcinoma cells cultured on a microporous support at the air/liquid interface. Reconstructed HVE were maintained in a humid atmosphere at 37°C with 5% CO2
. Hemalun/Eosin histological sections
generated from paraffin-embedded tissues revealed an early stratification after three days of culture that could be increased until day 14 with no visible sign of mortality (Figure 1). Cells in the different layers were non-keratinized squamous cells similar to the basal layer of native vaginal tissues. After 18 days in culture, apical cells displayed a morphology close to the native suprabasal layer could be detected (Figure 1). However, a loss of tissue architecture
with increased intercellular spaces was also observed, predicting a reduction in cell survival and a visible failure of the barrier function at
November 2025 PERSONAL CARE
ABSTRACT
In line with a growing demand for non- animal in vitro alternatives for the safety testing of feminine and cosmetic care products, the present study describes the in- house reconstruction of a vaginal organoid tissue model and demonstrates its use to evaluate topical ingredients commonly found in feminine care formulations. The 3D human vaginal epithelium (HVE) in vitro model reconstructed from isolated vulvar epithelial cells displayed in vivo- like structure and functionality from batch to batch, supporting its use as a reliable and reproducible in vitro tool to evaluate the safety and efficacy of intimate care ingredients and final products
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