Objectives The diagnosis of vocal fold cancer currently relies on invasive surgical biopsies, which can compromise laryngeal function. Distinguishing between different types of laryngeal lesions without invasive tissue sampling is therefore crucial. Autofluorescence spectroscopy (AFS) has proved to be efficient as a non-invasive detection technique but has yet to be fully exploited in the context of a multi-class tissue analysis. This study evaluates whether AFS can be used to discriminate between different types of laryngeal lesions in view of assisting in vocal fold surgery and preoperative investigations.

Materials and methods Ex vivo spectral autofluorescence scans were recorded for each sample using a 405-nm laser excitation. A total of 1308 spectra were recorded from 29 vocal fold samples obtained from 23 patients. Multiclass analysis was conducted on the spectral data, classifying lesions either as normal, benign, dysplastic, or carcinoma. The results were compared to histopathological diagnosis.

Results Through an appropriate selection of spectral components and a cascading classification approach based on artificial neural networks (ANN), a classification rate of 97% was achieved for each lesion class, compared to 52% using autofluorescence intensity.

Conclusion The study demonstrates the effectiveness of AFS combined with multivariate analysis for accurate classification of vocal fold lesions. Comprehensive spectral data analysis significantly improves classification accuracy, even in challenging situations such as distinguishing between malignant and premalignant or benign lesions. This method could provide a way to perform in situ mapping of tissue states for minimally-invasive biopsy and surgical resection margins.

The authors have declared no competing interest.

NCT03585075

This study was funded by the European Union Seventh Framework Programme FP7 2007 2013 undergrant number 288663 and Agence Nationale de la Recherche (ANR 17 CE190005)

I confirm all relevant ethical guidelines have been followed, and any necessary IRB and/or ethics committee approvals have been obtained.

Yes

The details of the IRB/oversight body that provided approval or exemption for the research described are given below:

The French ethical committee (Comite de Protection des Personnes (CPP) - EST II) gave ethical approval for this work.

I confirm that all necessary patient/participant consent has been obtained and the appropriate institutional forms have been archived, and that any patient/participant/sample identifiers included were not known to anyone (e.g., hospital staff, patients or participants themselves) outside the research group so cannot be used to identify individuals.

Yes

I understand that all clinical trials and any other prospective interventional studies must be registered with an ICMJE-approved registry, such as ClinicalTrials.gov. I confirm that any such study reported in the manuscript has been registered and the trial registration ID is provided (note: if posting a prospective study registered retrospectively, please provide a statement in the trial ID field explaining why the study was not registered in advance).

Yes

I have followed all appropriate research reporting guidelines, such as any relevant EQUATOR Network research reporting checklist(s) and other pertinent material, if applicable.

Yes

All data produced in the present study are available upon reasonable request to the authors