Safety and performance analysis of acriflavine and methylene blue for in vivo imaging of precancerous lesions using fibered confocal fluorescence microscopy (FCFM): an experimental study

BMC Pulmonary Medicine

Luc Thiberville¹  François-Xavier Boland⁴  Pierre Bohn⁵  Richard Sesboüé²  Liana Veresezan³  Mathieu Salaun¹  Bérengère Obstoy⁵ 

[1] Clinique Pneumologique and CIC INSERM U1404, IRIB, Rouen University Hospital, Rouen F-76031, France;INSERM U-1079, IRIB, Rouen University, Rouen F-76000, France;Department of Pathology, H. Becquerel Cancer Center, Rouen, France;Laboratory of Experimental Surgery, EA 3830 GRHV, IRIB, Rouen University, Rouen F-76000, France;Quant.I.F Litis EA 4108, IRIB, Rouen University, Rouen F-76000, France

关键词: Precancerous lesions;    Methylene blue;    Heller model;    Genotoxicity;    Fibered confocal fluorescence microscopy;    Diagnosis;   

Others  :  1170570
DOI  :  10.1186/s12890-015-0020-4

 received in 2014-11-01, accepted in 2015-03-10,  发布年份 2015

【 摘 要 】

Background

Fibered confocal fluorescence microscopy (FCFM) allows in vivo investigation of pulmonary microstructures. However, the bronchial epithelium can only be imaged using exogenous fluorophores. The objective of this study is to compare methylene blue (MB) and acriflavine genotoxicity and to assess FCFM performance for in vivo imaging of precancerous lesions.

Methods

Genotoxicity was assessed using the comet assay on both cultured human lymphocytes and NCI-H460 cells, which had been exposed to MB or acriflavine before being illuminated at 660 or 488 nm, respectively. FCFM was performed on precancerous lesions in the hamster cheek pouch model, following topical application of the fluorophores. FCFM data were analyzed according to histology.

Results

No genotoxicity was found using 0.01% (w/v) MB after illumination at 660 nm for 2 and 15 min (5 mW). Acriflavine exposure (0.025%) led to DNA damages, increasing from 2 to 15 min of light exposure at 448 nm in both lymphocytes (83.4 to 88%, p = 0.021) and NCI H460 cell populations (79.9 to 84.6%, p = 0.045). In total, 11 invasive carcinoma, 24 reserve cell hyperplasia, and 17 dysplasia lesions were imaged using FCFM in vivo. With both fluorophores, the cellular density increased from hyperplasia to high-grade dysplasia (p < 0.05). With MB, the cellular diameter significantly decreased (48.9 to 13.9 μm) from hyperplasia to carcinoma (p < 0.05). In this model, a cut-off diameter of 30 μm enabled the diagnosis of high-grade lesions with a sensitivity of 94.7% and a specificity of 97%.

Conclusion

Methylene blue can be used safely to image precancerous lesions in vivo. This study does not support the use of acriflavine in humans.

【 授权许可】

   
2015 Obstoy et al.; licensee BioMed Central.

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