title: Recellularizing of human acellular dermal matrices imaged by high-definition optical coherence tomography.
abstract: High-definition optical coherence tomography (HD-OCT) permits real-time 3D imaging of the impact of selected agents on human skin allografts. The real-time 3D HD-OCT assessment of (i) the impact on morphological and cellular characteristics of the processing of human acellular dermal matrices (HADMs) and (ii) repopulation of HADMs in vitro by human fibroblasts and remodelling of the extracellular matrix by these cells. Four different skin decellularization methods, Dispase II/Triton X-100, Dispase II/SDS (sodium dodecyl sulphate), NaCl/Triton X-100 and NaCl/SDS, were analysed by HD-OCT. HD-OCT features of epidermal removal, dermo-epidermal junction (DEJ) integrity, cellularity and dermal architecture were correlated with reflectance confocal microscopy (RCM), histopathology and immunohistochemistry. Human adult dermal fibroblasts were in vitro seeded on the NaCl/Triton X-100 processed HADMs, cultured up to 19 days and evaluated by HD-OCT in comparison with MTT proliferation test and histology. Epidermis was effectively removed by all treatments. DEJ was best preserved after NaCl/Triton X-100 treatment. Dispase II/SDS treatment seemed to remove all cellular debris in comparison with NaCl/Triton X-100 but disturbed the DEJ severely. The dermal micro-architectural structure and vascular spaces of (sub)papillary dermis were best preserved with the NaCl/Triton X-100. The impact on the 3D structure and vascular holes was detrimental with Dispase II/SDS. Elastic fibre fragmentation was only observed after Dispase II incubation. HD-OCT showed that NaCl/Triton X-100 processed matrices permitted in vitro repopulation by human dermal fibroblasts (confirmed by MTT test and histology) and underwent remodelling upon increasing incubation time. Care must be taken in choosing the appropriate processing steps to maintain selected properties of the extracellular matrix in HADMs. Processing HADMs with NaCl/Triton X-100 permits in vitro the proliferation and remodelling activity of human dermal fibroblasts. HD-OCT provides unique real-time and non-invasive 3D imaging of tissue-engineered skin constructs and complementary morphological and cytological information.
authors.affiliation: Department of Dermatology, Hôpital Erasme, Université Libre de Bruxelles, Brussels, Belgium.
authors.forename: Marc A L M
authors.initials: MA
authors.lastname: Boone
authors.forename: Jean Pierre
authors.initials: JP
authors.lastname: Draye
authors.forename: Gunther
authors.initials: G
authors.lastname: Verween
authors.forename: Annalisa
authors.initials: A
authors.lastname: Aiti
authors.forename: Jean-Paul
authors.initials: JP
authors.lastname: Pirnay
authors.forename: Gilbert
authors.initials: G
authors.lastname: Verbeken
authors.forename: Daniel
authors.initials: D
authors.lastname: De Vos
authors.forename: Thomas
authors.initials: T
authors.lastname: Rose
authors.forename: Serge
authors.initials: S
authors.lastname: Jennes
authors.forename: Gregor B E
authors.initials: GB
authors.lastname: Jemec
authors.forename: Veronique
authors.initials: V
authors.lastname: Del Marmol
mesh_terms: D062088:Acellular Dermis; D000328:Adult; D049109:Cell Proliferation; D002478:Cells, Cultured; D003199:Computer Systems; D020405:Dermis / Q000166:cytology; D005347:Fibroblasts / Q000166:cytology; D006801:Humans; D021621:Imaging, Three-Dimensional; D017830:Octoxynol; D016038:Skin Transplantation; D012965:Sodium Chloride; D023822:Tissue Engineering; D054457:Tissue Scaffolds; D041623:Tomography, Optical Coherence / Q000379:methods*; D014184:Transplantation, Homologous
chemical_list: D012965:Sodium Chloride; D017830:Octoxynol
keywords: high-definition optical coherence tomography; human acellular dermal matrices; human adult fibroblasts; processing; recellularising