505. EVALUATION OF PRO-INFLAMMATORY BIOMARKERS OF IRRITATION IN HUMAN SKIN MODEL USING MULTIPLEXED IMMUNOASSAY.

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Izbicka¹, E., Yee², B., Zhang², A., Streeper¹, R.T. ¹BTNS LLC, San Antonio, TX, USA and ²Panomics Inc., Fremont, CA, USA.

Summary:
This study by researchers at BTNS LLC and Panomics Inc. demonstrated that MatTek's EpiDerm in vitro human skin tissue equivalent is an excellent in vitro model for analyzing the potential anti-inflammatory effects of formulations designed to be applied topically on human skin.

Objective: The study evaluated multiplexed immunoassays to identify panels of biomarkers for efficacy of novel therapeutics for dermatologic conditions in a preclinical ex vivo model.

Methods: We used human EpiDerm™ model to compare modulation of skin irritation due to a topical exposure to 1.5% croton oil (CO) as a single agent or concurrent with either 0.5% indomethacin or 25% HF1107, a novel fatty acid derivative (patent pending). Following 18-hour exposure, toxicity was assessed by MTT conversion. Levels of 39 human chemokines, cytokines, and PGE(2) in conditioned media and tissue lysates were measured using multiplexed Luminex immunoassays. Inter-group differences were assessed by Student t-test.

Results: Toxicity of HF1107 was comparable with that of indomethacin by MTT. Over 50% (21/40) of the examined analytes showed highly significant inter-group differences (p<0.0002). In comparison with CO, HF1107 alone downregulated 14 and upregulated 1 medium marker; 9 tissue markers were downregulated. Respective numbers for indomethacin alone were: medium (11 down) and tissue (3 down). Concurrent HF1107 and CO downregulated 10 and upregulated 3 medium markers; respective numbers for indomethacin were 12 down and 4 up. Interestingly, HF1107 plus CO downregulated 12 tissue markers versus 3 down and 5 up for indomethacin in the same regimen.

Conclusions: The results demonstrate superior anti-inflammatory activity of HF1107 versus indomethacin and support an extended multiplexed immunoassay-based analysis of skin biomarker profiles in the EpiDerm model as robust tools in development of novel agents for treatment of skin diseases and for analysis of irritancy of cosmetics and other agents.

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EpiDerm Reconstructed Human Epidermis (RhE) Data Sheet

EpiDerm Specifications

EpiDerm Technical References

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Applications: Biomarker research, Inflammation - skin, Irritation - skin

Keywords: Anti-inflammatory activity, Biomarker, Biomarkers, EPI-200, EpiDerm, FGF-2, IL-12, IL-1a, IL-1ra, IL-8, Luminex assays, MMP, MMP-1, MMP-13, Multiplexed Luminex immunoassays, PGE2, Pro-inflammatory biomarkers, Skin irritancy, Skin irritation, VEGF

Materials Tested: Croton oil, Dexamethasone, Dyphenylhydramine, Indomethacin

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