Contributions of cell subsets to cytokine production during normal and impaired wound healing

Abstract

The objective of this study was to determine the relative contributions of different cell subsets to the production of cytokines and growth factors during normal and impaired wound healing. Cells were isolated from wounds of non-diabetic and diabetic mice and separated by magnetic sorting into neutrophils/T cells/B cells (NTB cell subset), monocytes/macrophages (Mo/Mp subset) and non-leukocytic cells including keratinocyte/fibroblast/endothelial cells (KFE subset). On both per cell and total contribution bases, the Mo/Mp subset was the dominant producer of pro-inflammatory cytokines interleukin (IL)-1β, tumor necrosis factor (TNF)-α and IL-6 in both non-diabetic and diabetic mice and was a significant producer of vascular endothelial cell growth factor (VEGF)-A, insulin-like growth factor (IGF)-1 and transforming growth factor (TGF)-β1. The NTB subset was also a significant producer of TNF-α and IL-10 whereas the KFE subset contributed significant amounts of VEGF, IGF-1 and TGF-β1. Sustained production of pro-inflammatory cytokines and impaired production of healing-associated factors were evident in each subset in diabetic mice. These data will be useful for further experimental and modeling studies on the role of cell subsets in wound healing as well as for designing therapeutic strategies for improving healing.

CD301b+ Macrophages Are Essential for Effective Skin Wound Healing

“…Further study of CD301b + macrophages may reveal molecular mechanisms by which myeloid cells function in tissue repair. Furthermore, induction of obesity and/or diabetes (Seitz et al, 2010) in Mgl2 DTR/GFP mice will allow the investigation of how this myeloid subset contributes to diabetes-impaired skin repair, which displays defects in myeloid cells (Mirza and Koh, 2011, 2014; Mirza et al, 2013). Finally, because myeloid cells function in multiple tissues to control regeneration after injury, our identification of a role of CD301b + myeloid cells in skin repair may have broad implications for regeneration of many tissue types.…”

“…Early stage macrophages are similar to classically activated or the M1 macrophages described by others (Brancato and Albina, 2011; Ferrante and Leibovich, 2012). Throughout healing, macrophage phenotype changes as macrophage cell surface protein and cytokine messenger RNA expression changes (Auffray et al, 2009; Brancato and Albina, 2011; Ferrante and Leibovich, 2012; Mirza and Koh, 2014). As regeneration proceeds into midstage healing, CD11b + /F4-80 + /Ly6C hi macrophages decline, and the macrophage pool expresses mannose receptor (CD206), Fizz1, IL-10, transforming growth factor (TGF)-β1 and vascular endothelial growth factor (Daley et al, 2010; Mirza and Koh, 2014; Werner and Grose, 2003).…”

“…Throughout healing, macrophage phenotype changes as macrophage cell surface protein and cytokine messenger RNA expression changes (Auffray et al, 2009; Brancato and Albina, 2011; Ferrante and Leibovich, 2012; Mirza and Koh, 2014). As regeneration proceeds into midstage healing, CD11b + /F4-80 + /Ly6C hi macrophages decline, and the macrophage pool expresses mannose receptor (CD206), Fizz1, IL-10, transforming growth factor (TGF)-β1 and vascular endothelial growth factor (Daley et al, 2010; Mirza and Koh, 2014; Werner and Grose, 2003). These midstage macrophages, referred to as alternatively activated or M2 macrophages (Gordon, 2003; Martinez et al, 2008), mediate epithelial and dermal repair (Delavary et al, 2011; Knipper et al, 2015; Lucas et al, 2010).…”

Constitutive overexpression of periostin delays wound healing in mouse skin

“…Taking these findings together, we assume that down‐regulation of IL‐1β and TNFα expression is one of the underlying mechanisms for how wound healing is impaired in Postn ‐overexpressing mice. M1‐like macrophages and neutrophils are the principal sources of IL‐1β and TNFα during wound repair . It was recently reported that treating mouse intraperitoneal macrophages with high doses of recombinant periostin changes the characteristics of these macrophages, decreasing expression of M1 markers including IL‐1β .…”

The Role of Adipocytes in Tissue Regeneration and Stem Cell Niches

“…Not only do skin macrophages influence hair follicle activation (Castellana et al 2014), but unique macrophage phenotypes are associated with various phases of skin regeneration after injury (Daley et al 2010, Mirza & Koh 2014). Macrophages are also required for the involution of the MG epithelium that is associated with MG WAT expansion (O’Brien et al 2010).…”