C1 Inhibitor Administration Reduces Local Inflammation and Capillary Leakage, Without Affecting Long-term Wound Healing Parameters, in a Pig Burn Wound Model.

BACKGROUND: Burns induce a boost in local and systemic complement levels as well as immune cell infiltration in the burn wound, which may negatively affect wound healing. OBJECTIVE: In this study, the effects of long-term treatment with complement inhibitor C1 esterase inhibitor (C1inh) on post-burn inflammation and wound healing parameters were analyzed in time up to 60 days post-burn. METHODS: Burned pigs were treated either with or without C1inh up to 15 days post-burn. Burn wound biopsies and blood were collected at different time points up to 60 days post-burn. Thereafter, complement in blood as well as complement and immune cells in the wound, capillary leakage, necrosis, reepithelialization and wound contraction were quantified. RESULTS: No significant differences in complement C3 blood levels were observed at any time point between C1inh-treated and control pigs. In the wound, complement C4 levels were significantly lower in the C1inh group than in controls at day 3-6 and 21-30 post-burn. Similarly, C3 levels, neutrophil and macrophage infiltration in the wound were, although not statistically significant, reduced in C1inh-treated pigs at day 9-14 post-burn. No differences in lymphocyte infiltration in the wound were found between C1inh and control pigs. C1inh-treated pigs also showed reduced capillary leakage. Despite these effects, no significant differences in the long-term wound healing parameters necrosis, reepithelialization and wound contraction were observed between C1inh and control pigs. CONCLUSION: In pigs, 15 days of C1inh treatment after burn, leads to a reduction in local inflammation and capillary leakage in the burn wound without affecting long-term wound healing parameters.

MRI and Additive Manufacturing of Nasal Alar Constructs for Patient-specific Reconstruction.

Surgical reconstruction of cartilaginous defects remains a major challenge. In the current study, we aimed to identify an imaging strategy for the development of patient-specific constructs that aid in the reconstruction of nasal deformities. Magnetic Resonance Imaging (MRI) was performed on a human cadaver head to find the optimal MRI sequence for nasal cartilage. This sequence was subsequently used on a volunteer. Images of both were assessed by three independent researchers to determine measurement error and total segmentation time. Three dimensionally (3D) reconstructed alar cartilage was then additively manufactured. Validity was assessed by comparing manually segmented MR images to the gold standard (micro-CT). Manual segmentation allowed delineation of the nasal cartilages. Inter- and intra-observer agreement was acceptable in the cadaver (coefficient of variation 4.6-12.5%), but less in the volunteer (coefficient of variation 0.6-21.9%). Segmentation times did not differ between observers (cadaver P = 0.36; volunteer P = 0.6). The lateral crus of the alar cartilage was consistently identified by all observers, whereas part of the medial crus was consistently missed. This study suggests that MRI is a feasible imaging modality for the development of 3D alar constructs for patient-specific reconstruction.

Collagen bundle morphometry in skin and scar tissue: a novel distance mapping method provides superior measurements compared to Fourier analysis.

Histopathological evaluations of fibrotic processes require the characterization of collagen morphology in terms of geometrical features such as bundle orientation thickness and spacing. However, there are currently no reliable and valid techniques of measuring bundle thickness and spacing. Hence, two objective methods quantifying the collagen bundle thickness and spacing were tested for their reliability and validity: Fourier first-order maximum analysis and Distance Mapping, with the latter constituting a newly developed morphometric technique. Histological slides were constructed and imaged from 50 scar and 50 healthy human skin biopsies and subsequently analyzed by two observers to determine the interobserver reliability via the intraclass correlation coefficient. An intraclass correlation coefficient larger than 0.7 is considered as representing good reliability. The interobserver reliability for the Fourier first-order maximum and for the Distance Mapping algorithms, respectively, showed an intraclass correlation coefficient above 0.72 and 0.89. Additionally, we performed an assessment of validity in the form of responsiveness, in particular, demonstrating medium to excellent results via a calculation of the effect size, highlighting that both methods are sensitive enough to measure a treatment effect in clinical practice. In summary, two reliable and valid measurement methods were demonstrated for collagen bundle morphometry for the first time. Due to its superior reliability and more useful measures (bundle thickness and bundle spacing), Distance Mapping emerges as the preferred and more practical method. Nevertheless, in the future, both methods can be used for reliable and valid collagen morphometry of skin and scars, whereas further applications evaluating the quantitative microscopy of other fibrotic processes are anticipated.