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

方法验证以评估 CBCT 检查后颊粘膜细胞和唾液的体内细胞和亚细胞变化

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

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.

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.

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