Method validation to assess in vivo cellular and subcellular changes in buccal mucosa cells and saliva following CBCT examinations

Niels Belmans, Liese Gilles, Piroska Virag, Mihaela Hedesiu, Benjamin Salmon, Sarah Baatout, Stéphane Lucas, Reinhilde Jacobs, Ivo Lambrichts, Marjan Moreels

Research output: Contribution to journalArticlepeer-review

Abstract

Objectives: Cone-beam CT (CBCT) is a medical imaging technique used in dental medicine. However, there are no conclusive data available indicating that exposure to X-ray doses used by CBCT are harmless. We aim, for the first time, to characterize the potential age-dependent cellular and subcellular effects related to exposure to CBCT imaging. Current objective is to describe and validate the protocol for characterization of cellular and subcellular changes after diagnostic CBCT. Methods: Development and validation of a dedicated two-part protocol: 1) assessing DNA double strand breaks (DSBs) in buccal mucosal (BM) cells and 2) oxidative stress measurements in saliva samples. BM cells and saliva samples are collected prior to and 0.5 h after CBCT examination. BM cells are also collected 24 h after CBCT examination. DNA DSBs are monitored in BM cells via immunocytochemical staining for γH2AX and 53BP1. 8-oxo-7,8-dihydro- 2′-deoxyguanosine (8-oxo-dG) and total antioxidant capacity are measured in saliva to assess oxidative damage. Results: Validation experiments show that sufficient BM cells are collected (97.1 ± 1.4 %) and that γH2AX/53BP1 foci can be detected before and after CBCT examination. Collection and analysis of saliva samples, either sham exposed or exposed to IR, show that changes in 8-oxo-dG and total antioxidant capacity can be detected in saliva samples after CBCT examination. Conclusion: The DIMITRA Research Group presents a two-part protocol to analyze potential age-related biological differences following CBCT examinations. This protocol was validated for collecting BM cells and saliva and for analyzing these samples for DNA DSBs and oxidative stress markers, respectively.

Original languageEnglish
Article number20180428
Pages (from-to)20180428
JournalDentomaxillofacial Radiology
Volume48
Issue number6
DOIs
Publication statusPublished - 1 Jan 2019

Keywords

  • Buccal mucosal cells
  • Dental cone-beam CT
  • DNA Double strand breaks
  • Oxidative stress
  • Saliva
  • Mouth Mucosa/cytology
  • X-Rays
  • Humans
  • Female
  • Male
  • Cone-Beam Computed Tomography
  • Spiral Cone-Beam Computed Tomography

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