Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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Expression of proinflammatory genes in epidermal keratinocytes is regulated by the hydration status.

ABSTRACT: We hypothesized that treatment of the injured skin with occlusion limits TEWL and results in a phenotype of epithelial response that more closely resembles mucosa. Here we addressed whether different hydration conditions change gene expression patterns in epidermis using microarray study in rabbit partial-thickness incisional wound. Microarray on epidermis showed that global expression patterns of the genes in full occluded vs. non-occluded wounds are distinct. Many genes - including IL-1M-NM-2, IL-8, TNF-M-NM-1, and COX-2-upregulated in skin environment were also upregulated in non-occluded wounds. Simulating increased TEWL showed increased expression of proinflammatory genes in human ex vivo skin culture (HESC) and stratified keratinocytes. Microarray on epidermis showed that global expression patterns of the genes in full occluded vs. non-occluded wounds are distinct. Female New Zealand White rabbits (3-4 kg) were purchased from Covance (Princeton, NJ) and were acclimated for a minimum of 7 days prior to experiments. Ketamine (45 mg/kg) and xylazine (7 mg/kg) were injected intramuscularly as primary anesthesia method prior to the procedure. The procedure was performed while the animal was in full anesthesia with the vital sign checking every 15 min including heart beat, breath frequency, and body temperature. Buprenex (0.05 mg/kg) was applied by intradermal injection prior to the surgery and continued every 6-12 hours for 24 hours after surgery. The animals were all monitored closely for signs of discomfort post surgeries. Tissue harvests were performed following euthanasia. The superficial incisional grids on rabbit ear were created by using a double blade scalpel with 1 mm space between the two blades. A single layer of Tegaderm was used to cover the wounding areas on both right and left ears until the wounding areas were fully re-epithelialized (two days post wounding, POE0). Thereafter, the Tegaderm on right ear was completely removed (NOW) while four more layers of Tegaderms were applied on left ear wounds (FOW). Rabbit ear samples were harvested at POE0, POE3 and POE5.

ORGANISM(S): Oryctolagus cuniculus

SUBMITTER: Wei Xu

PROVIDER: E-GEOD-42653 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Bacterial infections of wounds are associated with poor healing and worse scarring. We sought to identify transcriptomic patterns associated with impaired healing of wounds infected with Klebsiella pneumoniae (K.p.) or Pseudomonas aeruginosa (P.a.) using a rabbit ear wound model. Wounds created on post-operative day (POD) 0 were infected on POD3, within the inflammatory phase of healing. On POD4 the infected wounds were harvested for microarray/transcriptome analysis. Other wounds with 24-hour infections were treated with topical antibiotic to promote biofilm formation and harvested on POD6 or POD12. On POD4 before antibiotic treatment, both wounds contained elevated transcripts that enriched predominantly into inflammation/infection-response pathways and functions characteristic of infiltrating leukocytes. But there were 5-fold more elevated transcripts in P.a.- than K.p.-infected wounds. Additionally, unique to P.a.-infected wounds, was a minor network of inflammation/infection-response molecules with predicted upstream regulation predominated by type I interferons. Also on POD4, Dnr-transcripts of both wounds were enriched into stress-response pathways such as EIF2 signaling. But there were 8-fold more Dnr-transcripts in P.a.- than K.p.-infected wounds, and many more of them enriched in the function, cell death, suggesting that resident dermal cells of P.a.-infected wounds failed to survive a more destructive P.a. infection. On POD6, following two days of antibiotic treatment, the biofilm-colonized wounds expressed magnitudes fewer inflammation and stress-response transcripts. However, a single regulatory network of P.a.-infected wounds was found to consist of Upr-transcripts enriching immune/infection-response functions predicted to be regulated by type I interferons, which was similar to the network unique to P.a.-infected wounds on POD4. On POD12, genes expressed by K.p.-infected wounds suggesting healing, while for P.a.-infected wounds they suggested stalled healing. The similarities and differences between the wound responses to these infections further define the molecular foundations of healing impaired by infections. Rabbit ear Wounds created on post-operative day (POD) 0 were infected with Klebsiella pneumoniae (K.p.) or Pseudomonas aeruginosa (P.a.) on POD3 and harvested on POD4 for RNA extraction. Other wounds with 24-hour infections were treated with topical antibiotic to promote biofilm formation and harvested on POD6 or POD12.

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Expression of proinflammatory genes in epidermal keratinocytes is regulated by the hydration status.

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