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A porcine model for pathomorphological age assessment of surgically excised skin wounds.

ABSTRACT: A porcine model of second intention wound healing was established and gross and histological changes needed for accurate assessing the age of wounds were determined. Twenty-five pigs were anesthetized before incision of four wounds on each. The wounds were left to heal from 1 h to 35 days when the pigs were euthanized. In 14 pigs, biopsies were sampled from two wounds between days 2 and 18. By histological evaluation the following characteristics were found useful for determining the age of wounds: neutrophil:macrophage ratio, angiogenesis, hyperplasia of fibroblasts, presence of hemosiderophages and granulation tissue. The latter was present from day 4 (n?=?8 wounds, 100%), but by gross evaluation it was not recognized until day 5 (n?=?4 wounds, 100%). From day 4 to 10, the thickness of granulation tissue increased by 1.2?±?2.4 mm/day. The thickness of collagen fibers within granulation tissue increased throughout the study period, and complete epithelization was reached from day 18.

SUBMITTER: Barington K

PROVIDER: S-EPMC5977753 | biostudies-literature | 2018 May

REPOSITORIES: biostudies-literature

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A porcine model for pathomorphological age assessment of surgically excised skin wounds.

Barington Kristiane K Dich-Jørgensen Kristine K Jensen Henrik Elvang HE

Acta veterinaria Scandinavica 20180530 1

A porcine model of second intention wound healing was established and gross and histological changes needed for accurate assessing the age of wounds were determined. Twenty-five pigs were anesthetized before incision of four wounds on each. The wounds were left to heal from 1 h to 35 days when the pigs were euthanized. In 14 pigs, biopsies were sampled from two wounds between days 2 and 18. By histological evaluation the following characteristics were found useful for determining the age of woun ...[more]

PMID: 29848347

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Project description:Due to the transepidermal potential of 15-50?mV, inside positive, an injury current is driven out of all human skin wounds. The flow of this current generates a lateral electric field within the epidermis that is more negative at the wound edge than at regions more lateral from the wound edge. Electric fields in this region could be as large as 40?mV/mm, and electric fields of this magnitude have been shown to stimulate human keratinocyte migration toward the wounded region. After flowing out of the wound, the current returns through the space between the epidermis and stratum corneum, generating a lateral field above the epidermis in the opposite direction. Here, we report the results from the first clinical trial designed to measure this lateral electric field adjacent to human skin wounds noninvasively. Using a new instrument, the Dermacorder®, we found that the mean lateral electric field in the space between the epidermis and stratum corneum adjacent to a lancet wound in 18-25-year-olds is 107-148?mV/mm, 48% larger on average than that in 65-80-year-olds. We also conducted extensive measurements of the lateral electric field adjacent to mouse wounds as they healed and compared this field with histological sections through the wound to determine the correlation between the electric field and the rate of epithelial wound closure. Immediately after wounding, the average lateral electric field was 122?±?9?mV/mm. When the wound is filled in with a thick, disorganized epidermal layer, the mean field falls to 79?±?4?mV/mm. Once this epidermis forms a compact structure with only three cell layers, the mean field is 59?±?5?mV/mm. Thus, the peak-to-peak spatial variation in surface potential is largest in fresh wounds and slowly declines as the wound closes. The rate of wound healing is slightly greater when wounds are kept moist as expected, but we could find no correlation between the amplitude of the electric field and the rate of wound healing.

Project description:This study demonstrates, for the first time, clinical testing of elongated silica microparticles (EMP) combined with tailorable nanoemulsions (TNE) to enhance topical delivery of hydrophobic drug surrogates. Likewise, this is the first report of 6-carboxyfluorescein (a model molecule for topically delivered hydrophobic drugs) AM1 & DAMP4 (novel short peptide surfactants) used in volunteers. The EMP penetrates through the epidermis and stop at the dermal-epidermal junction (DEJ). TNE are unusually stable and useful because the oil core allows high drug loading levels and the surface properties can be easily controlled. At first, we chose alginate as a crosslinking agent between EMP and TNE. We initially incorporated a fluorescent lipophilic dye, DiI, as a hydrophobic drug surrogate into TNE for visualization with microscopy. We compared four different coating approaches to combine EMP and TNE and tested these formulations in freshly excised human skin. The delivery profile characterisation was imaged by dye- free coherent anti-Stoke Raman scattering (CARS) microscopy to detect the core droplet of TNE that was packed with pharmaceutical grade lipid (glycerol) instead of DiI. These data show the EMP penetrating to the DEJ followed by controlled release of the TNE. Freeze-dried formulations with crosslinking resulted in a sustained release profile, whereas a freeze-dried formulation without crosslinking showed an immediate burst-type release profile. Finally, we tested the crosslinked TNE coated EMP formulation in volunteers using multiphoton microscopy (MPM) and fluorescence-lifetime imaging microscopy (FLIM) to document the penetration depth characteristics. These forms of microscopy have limitations in terms of image acquisition speed and imaging area coverage but can detect fluorescent drug delivery through the superficial skin in volunteers. 6-Carboxyfluorescein was selected as the fluorescent drug surrogate for the volunteer study based on the similarity of size, charge and hydrophobicity characteristics to small therapeutic drugs that are difficult to deliver through skin. The imaging data showed a 6-carboxyfluorescein signal deep in volunteer skin supporting the hypothesis that EMP can indeed enhance the delivery of TNE in human skin. There were no adverse events recorded at the time of the study or after the study, supporting the use of 6-carboxyfluorescein as a safe and detectable drug surrogate for topical drug research. In conclusion, dry formulations, with controllable release profiles can be obtained with TNE coated EMP that can effectively enhance hydrophobic payload delivery deep into the human epidermis.

Dataset Information

A porcine model for pathomorphological age assessment of surgically excised skin wounds.

Publications

A porcine model for pathomorphological age assessment of surgically excised skin wounds.

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