Oral epithelial cell reaction after exposure to Invisalign plastic material

Am J Orthod Dentofacial Orthop. 2014 Jan;145(1):64-71. doi: 10.1016/j.ajodo.2013.09.011.


Introduction: Invisalign plastic aligners (Align Technology, Santa Clara, Calif) are used to correct malocclusions. The aligners wrap around the teeth and are in contact with gingival epithelium during treatment. The purpose of this study was to evaluate the cellular responses of oral epithelium exposed to Invisalign plastic in vitro.

Methods: Oral epithelial cells were exposed to eluate obtained by soaking Invisalign plastic in either saline solution or artificial saliva for 2, 4, and 8 weeks. Cells grown in media containing saline solution or saliva served as controls. Morphologic changes were assessed by light microscopy. The 3-[4, 5-dimethythiazol- 2-yl]-2, 5-diphenyl tetrazolium bromide assay and flow cytometry were used to determine cell viability and membrane integrity, respectively. Cellular adhesion and micromotion of epithelial cells were measured in real time by electrical cell-substrate impedance sensing.

Results: Cells exposed to saline-solution eluate appeared rounded, were lifted from the culture plates, and demonstrated significantly increased metabolic inactivity or cell death (P <0.05). Saliva eluates did not induce significant changes in cell viability compared with untreated cells. Flow cytometry and electric cell-substrate impedance sensing showed that cells treated with saline-solution eluate exhibited compromised membrane integrity, and reduced cell-to-cell contact and mobility when compared with saliva-eluate treatment.

Conclusions: Exposure to Invisalign plastic caused changes in viability, membrane permeability, and adhesion of epithelial cells in a saline-solution environment. Microleakage and hapten formation secondary to compromised epithelial integrity might lead to isocyanate allergy, which could be systemic or localized to gingiva. However, these results suggest that saliva might offer protection.

Publication types

  • Comparative Study

MeSH terms

  • Cell Adhesion / drug effects
  • Cell Culture Techniques
  • Cell Death / drug effects
  • Cell Line
  • Cell Membrane / drug effects
  • Cell Membrane Permeability / drug effects
  • Cell Movement / drug effects
  • Cell Survival / drug effects
  • Colorimetry / methods
  • Coloring Agents
  • Culture Media
  • Dental Materials / chemistry
  • Dental Materials / pharmacology*
  • Electric Impedance
  • Epithelial Cells / drug effects
  • Flow Cytometry
  • Gingiva / cytology
  • Gingiva / drug effects*
  • Humans
  • Keratinocytes / drug effects*
  • Materials Testing
  • Orthodontic Appliances*
  • Plastics / chemistry
  • Plastics / pharmacology*
  • Polyurethanes / chemistry
  • Polyurethanes / pharmacology*
  • Saliva / physiology
  • Saliva, Artificial / chemistry
  • Sodium Chloride / chemistry
  • Tetrazolium Salts
  • Thiazoles
  • Time Factors
  • Tooth Movement Techniques / instrumentation*


  • Coloring Agents
  • Culture Media
  • Dental Materials
  • Plastics
  • Polyurethanes
  • Saliva, Artificial
  • Tetrazolium Salts
  • Thiazoles
  • Sodium Chloride
  • thiazolyl blue