Recellularizing of human acellular dermal matrices imaged by high-definition optical coherence tomography.

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.

Differentiating allergic and irritant contact dermatitis by high-definition optical coherence tomography: a pilot study.

Differentiation of allergic contact dermatitis (ACD) and irritant contact dermatitis (ICD) is important because of different management requirements. Various non-invasive tests have been used in an attempt to improve diagnosis. In irritant dermatitis, thickening of the epidermis has been a constant finding. High-Definition Optical Coherence Tomography (HD-OCT) is a non-invasive real-time three-dimensional imaging technique with cellular resolution for which an adapted algorithmic method for pattern analysis discriminating inflammatory skin diseases has been proposed. The aim of this study was threefold. (1) To evaluate the correlation between HD-OCT features and clinical scores of allergic and irritant patch test reactions. (2) To explore the potential of HD-OCT in optimizing the visual patch test scoring. (3) To assess in vivo the cytological and 3-D micro-architectural differences in skin reaction types between doubtful positive ACD and ICD. Twenty-two volunteers were patch tested using potassium(VI)dichromate, cobalt(II)chloride, nickel(II) sulfate and palladium(II)chloride. Visual patch test scoring and HD-OCT assisted patch test scoring were performed at 48 and 96 h after patch test application according to ECDRG guidelines. Selected HD-OCT features correlated well with clinical severity scores. HD-OCT assessment improved the visual patch test scoring although not significantly. Increased epidermal thickness observed in ICD at first reading was a significant finding useful in differentiating doubtful (+?) ACD from irritant (IR) ICD reactions. In conclusion, HD-OCT might be a unique tool for in vivo non-invasive real-time three-dimensional epidermal thickness measurements helping to differentiate IR from doubtful (+?) reactions in patch testing. Selected HD-OCT features corresponded well with severity of visual scoring. These features might help to quantify the degree of inflammation in inflammatory skin conditions. HD-OCT might help in optimizing visual patch test scoring in some situations.

High-definition optical coherence tomography imaging of melanocytic lesions: a pilot study.

High-definition optical coherence tomography (HD-OCT) is a non-invasive in vivo imaging technique with cellular resolution based on the principle of conventional optical coherence tomography. The objective of this study was to evaluate HD-OCT for its ability to identify architectural patterns and cytologic features of melanocytic lesions. All lesions were examined by one observer clinically and using dermoscopy. Cross-sectional HD-OCT images were compared with histopathology. En face HD-OCT images were compared with reflectance confocal microscopy (RCM). Twenty-six melanocytic lesions of 26 patients were imaged. Identification of architectural patterns in cross-sectional mode and cytologic features of pigmented cells in the epidermis, dermo-epidermal junction, papillary dermis, and superficial reticular dermis in the en face mode was possible by HD-OCT. HD-OCT provides morphological imaging with sufficient resolution and penetration depth to discriminate architectural patterns and cytologic features of pigmented cells in epidermis and dermis. The method appears to offer the possibility of additional three-dimensional structural information complementary to that of RCM, albeit at a slightly lower lateral resolution. The diagnostic potential of HD-OCT regarding malignant melanoma is not high enough for ruling out a diagnosis of malignant melanoma.

Imaging actinic keratosis by high-definition optical coherence tomography. Histomorphologic correlation: a pilot study.

With the continued development of non-invasive therapies for actinic keratosis such as PDT and immune therapies, the non-invasive diagnosis and monitoring become increasingly relevant. High-definition optical coherence tomography is a high-resolution imaging tool, with micrometre resolution in both transversal and axial directions, enable to visualize individual cells up to a depth of around 570 µm filling the imaging gap between conventional optical coherence tomography and reflectance confocal microscopy. We sought to determine the feasibility of detecting and grading of actinic keratosis by this technique using criteria defined for reflectance confocal microscopy compared to histology. In this pilot study, skin lesions of 17 patients with a histologically proven actinic keratosis were imaged by high-definition optical coherence tomography just before excision and images analysed qualitatively. The surrounding normal looking skin has been used as control group. In lesional skin, dyskeratotic and atypical keratinocytes could be noticed with this new technique. An atypical honeycomb pattern in variable degree or a disarranged epidermal pattern could be observed. A good correlation between the dimension of atypia and/or disarrangement of the spinous-granular layer on en face images and the histopathological grading could be demonstrated. Relevant cross-sectional imaging criteria could be defined for the different histopathological variants of actinic keratoses. The surrounding skin displayed features of photodamage. Using features already suggested by reflectance confocal microscopy, the study implies that high-definition optical coherence tomography facilitates in vivo diagnosis of actinic keratosis and allows the grading of different actinic keratosis lesions for increased clinical utility.