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1.
J Tissue Eng Regen Med ; 11(3): 713-723, 2017 03.
Article in English | MEDLINE | ID: mdl-25546487

ABSTRACT

An FDA-approved, prototypic, living, bilayered skin construct (BSC) has been used for non-healing wounds. Using this particular construct as proof of principle, we hypothesized that an in vitro 'priming' step may enhance its repertoire of expression of key mediators and genes. The priming step used here was incubation in Dulbecco's modified Eagle's medium (DMEM) for 24 h at 37°C and 5% CO2 , with or without construct meshing. Microarray and ingenuity pathway analysis (IPA) showed that >1000 genes were overexpressed by the priming step, including interleukin 6 (IL-6), which plays important roles in wound healing. Genes highly overexpressed by priming were those involved in epidermal proliferation and migration. Quantitative real-time PCR (qRT-PCR), immunostaining and western blots verified the results. An epiboly assay (epidermal migration over dermis) showed that BSC epiboly was inhibited by IL-6 neutralizing antibody. Back wounds of nude mice were treated with primed or control BSCs for 3 days prior to harvesting; primed BSCs showed a significantly (p = 0.006) greater level of epidermal migration vs unprimed. Our study demonstrates that an in vitro priming step induces wound healing-related genes in the BSC, leading to a construct that could prove more effective in stimulating wound healing. Copyright © 2014 John Wiley & Sons, Ltd.


Subject(s)
Cell Movement , Epidermal Cells , Tissue Engineering/methods , Tissue Scaffolds/chemistry , Animals , Antibodies, Neutralizing/pharmacology , Cell Movement/drug effects , Cell Movement/genetics , Cell Proliferation/drug effects , Cluster Analysis , Interleukin-6/immunology , Keratin-17/metabolism , Mice, Nude , Oligonucleotide Array Sequence Analysis , Real-Time Polymerase Chain Reaction , Reproducibility of Results , Transcriptome
2.
Wound Repair Regen ; 16(4): 503-6, 2008.
Article in English | MEDLINE | ID: mdl-18638268

ABSTRACT

Dermal pericapillary fibrin is a hallmark of venous disease and is thought to play a pathogenic role in the development of ulceration. However, the actual spatial configuration of pericapillary fibrin is unknown, and it remains unclear whether it truly represents a barrier that can impair physiological exchanges between the blood and dermis. Using confocal microscopy on tissue specimens taken from the edges of venous ulcers in six patients, we report a detailed analysis of dermal pericapillary fibrin deposits. Sections were evaluated with an antibody to human fibrinogen/fibrin and viewed, vertically and horizontally, with confocal microscopy. The distribution of fibrin deposition was highly variable and patchy, with areas of great intensity next to others of marginal intensity. Vertical cut sections showed the highest concentration of fluorescent material next to the lumen of dermal capillaries. Horizontal sections showed that maximal fluorescence was distributed at random. Our findings indicate that fibrin deposits in venous ulcers are patchy and discontinuous around dermal vessels. As such, these deposits are unlikely to act as a true and stable anatomic barrier as originally proposed. However, pericapillary fibrin may still act as a physiological barrier under conditions of poor blood flow where even marginal or patchy fibrin deposition might have a greater effect on the exchange of oxygen and other nutrients between blood and dermis.


Subject(s)
Fibrin/metabolism , Skin/metabolism , Varicose Ulcer/metabolism , Adult , Aged , Aged, 80 and over , Biopsy , Female , Humans , Leg/blood supply , Male , Microscopy, Confocal , Middle Aged , Skin/blood supply , Skin/pathology , Varicose Ulcer/pathology
3.
J Food Prot ; 46(5): 400-402, 1983 May.
Article in English | MEDLINE | ID: mdl-30913645

ABSTRACT

Chlorinated, recycled water for cooling of containers in still retorts was sampled over a 27-month period at one food processing plant. Of 274 samples taken, 28 contained mesophilic, anaerobic spores in numbers that ranged from 0.04-4.6/ml (MPN). Though all isolates were characterized as Clostridium species, 11% could not be matched with named species. Clostridium butyricum and Clostridium barati (synonyms: C. paraperfringens , C. perenne ) comprised 55% of isolates. Excepting Clostridium sticklandii , which is neither proteolytic nor saccharolytic, all isolates were saccharolytic. This contrasted with the finding of both proteolytic and saccharolytic clostridial spores in the municipal water feeding the recycle water reservoir. An apparent selection for saccharolytic strains could not be explained on the basis of published resistance of anaerobic spores to free available chlorine.

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