Characterization of representative plasticizer exposures in NICU neonates

Characterization of representative plasticizer exposures in NICU neonates using non-negative matrix factorization: a data-driven approach to assess endocrine disruption risk in mixtures

S. Cambier 1, V. Sautou 2, L. Dahbi 3, P. Chennell 2, B. Pereira 1, M.-C. Chagnon 3, L. Bernard 2*

1 Clinical Research and Innovation Direction, CHU Clermont-Ferrand, F-63000 Clermont-Ferrand, France

2 ICCF UMR CNRS 6296, CHU de Clermont-Ferrand, Université Clermont Auvergne, France

3 University of Bourgogne Europe, INSERM CTM UMR 1231, Nutrition Physiology and Toxicology Laboratory (NUTOX), platform “Packtox” L'Institut Agro Dijon, 1 Esplanade Erasme, F-21000 Dijon, France

*Corresponding author: lise.bernard@uca.fr

Introduction / Context:

Plasticizers are widely used in flexible PVC medical devices. Among these compounds, DEHP is known to be an endocrine-disruptor. It has been progressively replaced by alternative plasticizers, such as DEHTP and TEHTM, which also exhibit toxic effects when considered individually at the levels of exposure observed in clinical settings (1–3). In neonatal intensive care units (NICUs), premature and critically ill newborns are exposed to complex and repeated mixtures of these compounds during medical care (4–8). The French ARMED-NEO biomonitoring study, funded by ANSM, has shown that newborns hospitalized are simultaneously exposed to multiple plasticizers, as evidenced by the detection of several urinary metabolites of DEHP and its alternatives (8). To assess the toxicological risk associated with such multiple exposure, it is essential to define representative exposure profiles as it is impossible to experimentally test all possible combinations of metabolites.

Materials and Methods:

Data on urinary metabolites from 97 neonates (508 samples) in the ARMED-NEO cohort were analyzed using non-negative matrix factorization (NMF), a mathematical representation that identifies latent mixture structures from complex data sets. Only concentrations above the limit of quantification were considered. The optimal number of mixtures was determined based on numerical stability (cophenetic stability) and residual error criteria.

Results and Discussion:

NMF identified three representative mixtures (Ma, Mb, Mc), with Ma (67%) dominated by 5-carboxy-MEHP, Mb (28%) by 5-carboxy-MEHTP, and Mc (2%) by DEHTM-1,4, reflecting exposures to DEHP and its substitutes. This approach provided a realistic, data-driven picture of multi-exposure in NICU neonates and allowed simplification of 23 metabolites into three interpretable profiles.

These mixtures now serve as the basis for targeted EATS (Estrogen, Androgen, Thyroid-and Steroidogenesis)-related hormonal in vitro bioassays in the NEOMIX project (funded by ANSES-PNR EST 2023), aiming to evaluate potential combined endocrine-disrupting activities and to investigate whether early-life exposure to these mixtures could be associated with neurodevelopmental outcomes.

Références:

1. Kambia N, Séverin I, Farce A, Dahbi L, Dine T, Moreau E, et al. Comparative Effects of Di-(2-ethylhexyl)phthalate and Di-(2-ethylhexyl)terephthalate Metabolites on Thyroid Receptors: In Vitro and In Silico Studies. Metabolites. 10 févr 2021;11(2):94.

2. Dahbi L, Farce A, Kambia N, Séverin I, Dine T, Moreau E, et al. In vitro and in silico approach to study the hormonal activities of the alternative plasticizer tri-(2-ethylhexyl) trimellitate TEHTM and its metabolites. Arch Toxicol. mars 2022;96(3):899‑918.

3. Kambia NK, Séverin I, Farce A, Moreau E, Dahbi L, Duval C, et al. In vitro and in silico hormonal activity studies of di-(2-ethylhexyl)terephthalate, a di-(2-ethylhexyl)phthalate substitute used in medical devices, and its metabolites. J Appl Toxicol JAT. juill 2019;39(7):1043‑56.

4. Panneel L, Cleys P, Poma G, Ait Bamai Y, Jorens PG, Covaci A, et al. Ongoing exposure to endocrine disrupting phthalates and alternative plasticizers in neonatal intensive care unit patients. Environ Int. avr 2024;186:108605.

5. Cleys P, Panneel L, Jorens PG, Mulder A, Covaci A. Neonatal Exposure to Phthalate and Alternative Plasticizers via Enteral Nutrition. Environ Sci Technol. 5 août 2025;59(30):15649‑60.

6. Stroustrup A, Bragg JB, Busgang SA, Andra SS, Curtin P, Spear EA, et al. Sources of clinically significant neonatal intensive care unit phthalate exposure. J Expo Sci Environ Epidemiol. janv 2020;30(1):137‑48.

7. Demirel A, Çoban A, Yıldırım Ş, Doğan C, Sancı R, İnce Z. Hidden Toxicity in Neonatal Intensive Care Units: Phthalate Exposure in Very Low Birth Weight Infants. J Clin Res Pediatr Endocrinol. 1 sept 2016;8(3):298‑304.

8. Bernard L, Masse M, Boeuf B, Chennell P, Decaudin B, Durand N, et al. Medical devices used in NICU: The main source of plasticisers’ exposure of newborns. Sci Total Environ. 1 févr 2023;858(Pt 3):159994.

Keywords: Medical devices – endocrine disruptors – neonatal multiexposure – risk assessment – dimension reduction