Premium
Real‐time three‐dimensional imaging of epidermal splitting and removal by high‐definition optical coherence tomography
Author(s) -
Boone Marc,
Draye Jean Pierre,
Verween Gunther,
Pirnay JeanPaul,
Verbeken Gilbert,
De Vos Daniel,
Rose Thomas,
Jennes Serge,
Jemec Gregor B. E.,
Del Marmol Véronique
Publication year - 2014
Publication title -
experimental dermatology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.108
H-Index - 96
eISSN - 1600-0625
pISSN - 0906-6705
DOI - 10.1111/exd.12516
Subject(s) - lamina densa , optical coherence tomography , dispase , biomedical engineering , epidermis (zoology) , confocal microscopy , dermoepidermal junction , materials science , basement membrane , chemistry , pathology , dermis , biology , anatomy , medicine , ophthalmology , microbiology and biotechnology , biochemistry , collagenase , enzyme
While real‐time 3‐ D evaluation of human skin constructs is needed, only 2‐ D non‐invasive imaging techniques are available. The aim of this paper is to evaluate the potential of high‐definition optical coherence tomography ( HD ‐ OCT ) for real‐time 3‐ D assessment of the epidermal splitting and decellularization. Human skin samples were incubated with four different agents: D ispase II , NaCl 1 M , sodium dodecyl sulphate ( SDS ) and T riton X ‐100. Epidermal splitting, dermo‐epidermal junction, acellularity and 3‐ D architecture of dermal matrices were evaluated by H igh‐definition optical coherence tomography before and after incubation. Real‐time 3‐ D HD ‐ OCT assessment was compared with 2‐ D en face assessment by reflectance confocal microscopy ( RCM ). (Immuno) histopathology was used as control. HD ‐ OCT imaging allowed real‐time 3‐ D visualization of the impact of selected agents on epidermal splitting, dermo‐epidermal junction, dermal architecture, vascular spaces and cellularity. RCM has a better resolution (1 μ m) than HD ‐ OCT (3 μ m), permitting differentiation of different collagen fibres, but HD ‐ OCT imaging has deeper penetration (570 μ m) than RCM imaging (200 μ m). Dispase II and N a C l treatments were found to be equally efficient in the removal of the epidermis from human split‐thickness skin allografts. However, a different epidermal splitting level at the dermo‐epidermal junction could be observed and confirmed by immunolabelling of collagen type IV and type VII . Epidermal splitting occurred at the level of the lamina densa with dispase II and above the lamina densa (in the lamina lucida) with NaCl . The 3‐ D architecture of dermal papillae and dermis was more affected by D ispase II on HD ‐ OCT which corresponded with histopathologic (orcein staining) fragmentation of elastic fibres. With SDS treatment, the epidermal removal was incomplete as remnants of the epidermal basal cell layer remained attached to the basement membrane on the dermis. With T riton X ‐100 treatment, the epidermis was not removed. In conclusion, HD ‐ OCT imaging permits real‐time 3‐ D visualization of the impact of selected agents on human skin allografts.