Norbert Kiss1,2, Kende Lőrincz1, Dóra Haluszka1,2, Enikő Kuroli1, Judit Hársing 1, Balázs Mayer1, Sarolta Kárpáti 1, György Fekete 3, Róbert Szipőcs2,4, Norbert Wikonkál1 and Márta Medvecz1
1 Department of Dermatology, Venereology and Dermatooncology, Semmelweis University, Budapest, Hungary
2 Wigner RCP, Institute for Solid State Physics and Optics, Budapest, Hungary
3 2nd Department of Pediatrics, Semmelweis University, Budapest, Hungary.
4 R&D Ultrafast Lasers Ltd, P.O. Box 622, H-1539 Budapest, Hungary
Introduction: Ehlers–Danlos syndrome (EDS) is the name for a heterogeneous group of rare genetic connective tissue disorders with an overall incidence of 1 in 5000. The classification of EDS recently has undergone significant changes, yet the description of the histological features of collagen morphology described in different EDS subtypes has endured the test of time since the 1980s. Nonlinear optical microscopy (NLO) techniques can be utilized for non-invasive in vivo label-free imaging of the skin.
Aims: As there is a niche of noninvasive diagnostic techniques of EDS, we set out to perform NLO imaging of EDS-affected skin.
Methods: Molecular genetic testing for EDS was carried out in all patients. Among NLO techniques two-photon absorption fluorescence (TPF) microscopy can visualize endogenous fluorophores, such as elastin, while the morphology of collagen fibers can be assessed by second-harmonic generation (SHG) microscopy. We performed TPF and SHG microscopy imaging on ex vivo skin samples of one patient with classical EDS and two patients with vascular EDS and two healthy controls. For histological analysis, hematoxylin and eosin (H&E), Van Gieson’s (VG) and Weigert's elastic (WE) stained sections were prepared.
Results: The genetic background of EDS was identified in all cases. We detected irregular, loosely dispersed collagen fibers in a non-parallel arrangement in the dermis of the EDS patients, while as expected, there was no noticeable impairment in the elastin content. These findings were also verified by the histopathologic reading of the H&E, VG and WE stained histology slides.
Conclusion: Based on our study, in vivo NLO imaging could be utilized for the non-invasive diagnostic testing of EDS and the assessment of the skin status of EDS patients in the future. Also, using NLO imaging it could be investigated how the dermal status relates to clinical outcomes in EDS patients.
Doctoral School: Clinical Medicine
Program: Dermatology and Venereology
Supervisor: Norbert Wikonkál
E-mail address: norbert.f.kiss @gmail.com