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Related Experiment Video

Updated: Apr 8, 2026

Author Spotlight: 3D Movement Assessment of Maxillary Posterior Teeth in Clear Aligner Treatment
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A Dual Cell-Line Molecular Platform to Assess Sub-Threshold Biocompatibility: Application to Processed Orthodontic

Katia Barbaro1, Ettore Candida2, Francesca Silvestrini Biavati2

  • 1Istituto Zooprofilattico Sperimentale Del Lazio e Della Toscana "M. Aleandri", Rome, Katia Barbaro, Italy.

Orthodontics & Craniofacial Research
|April 7, 2026
PubMed
Summary

Orthodontic clear aligners show minimal immune response, even when trimmed, confirming their safety for oral use. This study utilized a sensitive dual-cell model to detect sub-threshold molecular effects, ensuring biocompatibility below toxicity levels.

Keywords:
biocompatibilityhormone receptorsinflammatory markersorthodontic clear alignerspolyethylene terephthalate glycol‐modified (PETG)

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Area of Science:

  • Biomaterials Science
  • Immunology
  • Endocrinology

Background:

  • Traditional biocompatibility testing often misses subtle molecular responses below toxicity thresholds, critical for long-term tissue contact materials.
  • Orthodontic clear aligners require thorough evaluation due to prolonged intraoral use and varying manufacturing methods.

Purpose of the Study:

  • To assess low-level hormonal and immune responses to orthodontic clear aligners using a dual-cell model (MCF-7 and THP-1).
  • To evaluate the biocompatibility of clear aligners manufactured via CNC milling or laser cutting.

Main Methods:

  • Utilized MTT assay for cell viability and RT-PCR for gene expression analysis of inflammatory markers (COX-2, IL-6, IL-8) and hormone receptors (ERα, ERβ).
  • Employed a dual-cell system (MCF-7, THP-1) to detect sub-threshold molecular responses.

Main Results:

  • Modest COX-2 modulation observed in THP-1 cells with milled aligners, without significant induction of IL-6 or IL-8.
  • No changes in estrogen receptor (ERα/ERβ) expression were detected.
  • All observed responses were below established toxicity thresholds for xenoestrogens and inflammatory mediators.

Conclusions:

  • The MCF-7 and THP-1 cell pair serves as a sensitive preclinical system for detecting sub-threshold bioactivity in dental materials.
  • Orthodontic clear aligners, including those trimmed post-manufacturing, do not exhibit antigenic properties, supporting their safe intraoral application.