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Migration inhibitory factor-related protein (MRP)8 and MRP14 are differentially expressed in free-electron laser and
Nanjun Wu1, Jeffrey M Davidson
1Department of Pathology, Vanderbilt University School of Medicine, Medical Center, Nashville, TN 37232-2561, USA.
Abstract:
Incisions made in mouse skin by scalpel or the free-electron laser heal at different rates. To identify genes that are differentially expressed in free-electron laser or scalpel wounds, we isolated total RNA from free-electron laser- or scalpel-produced incisions and normal skin at day 7 postwounding. cDNA microarray analysis identified 89 of 15,000 genes in a mouse microarray as having significantly different expression levels. Migration inhibitory factor-related protein (MRP) 14 was almost 30 times more highly expressed in scalpel wounds than in free-electron laser wounds. This result was confirmed by Northern blot analysis, which also showed that scalpel wounds expressed higher levels of MRP8, a related S100 protein that can heterodimerize with MRP14, at days 2, 7, and 14 postwounding. Free-electron laser wounds also showed elevated expression of MRP8 and MRP14 relative to normal skin. In situ hybridization showed that the patterns of MRP14 and MRP8 expression in free-electron laser and scalpel wound tissues were similar. MRP14 and MRP8 were expressed in the dermal wound margin, while a very low level of MRP14 and MRP8 expression was seen in the migrating epidermis. Dual immunofluorescence staining for MRP14 or MRP8 and macrophage (F4/80) showed that most of the wound macrophages simultaneously expressed MRP14 and MRP8. Some expression was also found in neutrophils, while neither antigen accumulated to a significant degree in the epidermis. Relatively lower MRP8 and 14 expression in free-electron laser wounds was correlated with a higher level of matrix metalloproteinase-13 expression and a reduced rate of wound healing. While the regulation of MRP8 expression in mouse may be different from human skin, we suggest that elevated expression of MRP8 and MRP14 may have a relevant therapeutic effect against inflammation in wound healing.
Insights
Scalpel wounds show higher expression of migration inhibitory factor-related proteins (MRP) 8 and 14 compared to free-electron laser wounds. Elevated MRPs in scalpel wounds correlate with slower healing, suggesting a therapeutic role in wound inflammation.
Area of Science:
- Wound healing research
- Molecular biology
- Dermatology
Background:
- Incisions from scalpels and free-electron lasers heal at different rates in mouse skin.
- Understanding gene expression differences is key to explaining varied healing rates.
Purpose of the Study:
- To identify genes with differential expression between free-electron laser and scalpel wounds.
- To investigate the role of Migration Inhibitory Factor-Related Proteins (MRP) 8 and 14 in wound healing.
Main Methods:
- RNA isolation from wound tissues and normal skin.
- cDNA microarray analysis to identify differentially expressed genes.
- Northern blot, in situ hybridization, and immunofluorescence staining to confirm and localize gene expression.
Main Results:
- 89 out of 15,000 genes showed significant expression differences.
- MRP14 was ~30 times higher in scalpel wounds vs. laser wounds.
- MRP8 and MRP14 were elevated in both wound types compared to normal skin, primarily in dermal macrophages and neutrophils.
Conclusions:
- Lower MRP8 and MRP14 expression in free-electron laser wounds correlated with higher matrix metalloproteinase-13 and reduced healing rates.
- Elevated MRP8 and MRP14 may offer therapeutic benefits against wound inflammation.
