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Enhanced Sensitivity and Human Relevance: a TNF-α-Induced HaCaT Keratinocyte Model for Screening Anti-Inflammatory
Weizuo Liao1, Yuxuan Xiao1, Qunying Leng2
1Food Safety and Health Research Center, NMPA Key Laboratory for Safety Evaluation of Cosmetics, Guangdong-Hongkong-Macao Joint Laboratory for Contaminants Exposure and Health, Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Public Health, Southern Medical University, Guangzhou, China.
A new TNF-α-induced inflammatory model using human HaCaT cells offers a sensitive, in vitro alternative for cosmetic ingredient screening. This human-relevant method reliably detects anti-inflammatory effects and distinguishes actives from controls.
Area of Science:
- In vitro toxicology and safety assessment
- Dermatology and cosmetic science
- Cell biology and immunology
Background:
- Conventional methods for evaluating cosmetic ingredient anti-inflammatory potential often rely on animal models or less sensitive cell lines.
- There is a growing need for reliable, human-relevant, and non-animal alternative methods in cosmetic safety and efficacy testing.
- Tumor Necrosis Factor-alpha (TNF-α) is a key mediator of inflammation relevant to skin conditions.
Purpose of the Study:
- To develop and validate a TNF-α-induced inflammatory model using human keratinocytes (HaCaT cell line) as an alternative in vitro method.
- To assess the sensitivity and reproducibility of this novel model for screening cosmetic ingredients.
- To compare the performance of the HaCaT model with the conventional lipopolysaccharide (LPS)-stimulated RAW264.7 murine macrophage model.
Main Methods:
- Development of a TNF-α-induced inflammatory model in HaCaT cells.
- Evaluation of anti-inflammatory potential of cosmetic ingredients by measuring interleukin-6 (IL-6) suppression.
- Comparison with the LPS-stimulated RAW264.7 macrophage model and multi-laboratory verification with a novel ingredient (β-nicotinamide mononucleotide, NMN).
Main Results:
- The TNF-α-HaCaT model demonstrated higher sensitivity, detecting significant cytokine inhibition at concentrations up to 1000-fold lower than the RAW264.7 model for certain actives.
- The model successfully distinguished known anti-inflammatory agents (ectoine, troxerutin, dipotassium glycyrrhizinate) from non-active controls (glycerin, butanediol, propylene glycol).
- The HaCaT model identified a potential pro-inflammatory effect at high concentrations of some ingredients, an observation not seen in the murine model. Multi-laboratory testing confirmed NMN's anti-inflammatory activity and assay reproducibility.
Conclusions:
- The TNF-α-HaCaT model is a sensitive, human-relevant, and reproducible alternative for screening the anti-inflammatory potential of cosmetic ingredients.
- This model offers advantages over conventional methods, including enhanced sensitivity and the ability to detect concentration-dependent pro-inflammatory shifts.
- The validated TNF-α-HaCaT assay contributes to the development of non-animal testing strategies for cosmetic safety and efficacy assessment.
