Short CommunicationContributions of cell subsets to cytokine production during normal and impaired wound healing
Introduction
Skin wound healing involves a series of overlapping events involving hemostasis, inflammation, new tissue formation and remodeling. A number of cell subsets contribute to healing, including keratinocytes, fibroblasts, endothelial cells and inflammatory cells [1], [2], [3], [4]. The activity of these cells is regulated by cytokines acting in both autocrine and paracrine fashion to bring about efficient healing. In the setting of diabetes, impaired healing is associated with persistent inflammation, reduced angiogenesis and granulation tissue formation, and impaired closure [5], [6], [7], [8], [9], [10]. These defects are associated with persistent production of pro-inflammatory cytokines and reduced release of pro-angiogenic and pro-healing factors.
A number of studies have reported that multiple cellular sources may contribute to cytokine production in wounds. The majority of these studies involve either immunohistochemical assessment of tissue sections or cell culture studies using primary cells or cell lines [1], [3], [4], [11]. However, these methods are not optimal for determining the relative contribution of cell subsets to wound cytokine levels. The objective of this study was to isolate cells directly from wounds and measure cytokine release from these cells ex vivo to establish the relative contributions of different cell subsets to the production of cytokines and growth factors during normal and impaired wound healing.
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Animals
Non-diabetic db/+ and diabetic db/db mice on a C57Bl/6 background were obtained from The Jackson Laboratory (Bar Harbor, ME). Experiments were performed on 12–16 week-old mice. All experimental procedures were approved by the Animal Care Committee at the University of Illinois at Chicago.
Excisional wounding
Each mouse was anesthetized with an intraperitoneal injection of ketamine (100 mg/kg) and xylazine (5 mg/kg) and its dorsum was shaved and cleaned with betadine and then alcohol swab. Four 8 mm excisional wounds
Results and discussion
Wound cells were analyzed on days 5 and 10 following excisional wounding, which correspond to the inflammatory and proliferative phases, respectively, in non-diabetic mice. In non-diabetic mice, Mo/Mp were present in the largest numbers on day 5, whereas KFE cells predominated on day 10 (Fig. 1a). In diabetic mice, the NTB and Mo/Mp cell subsets were present at similar levels which were higher than that of the KFE subset on both days 5 and 10, demonstrating the inability to progress through the
Acknowledgments
This study was supported by the National Institutes of Health (R01GM092850). The authors thank Dr. Luisa DiPietro, University of Illinois at Chicago, for critical comments on a previous draft of this manuscript.
References (15)
- et al.
Inflammation in wound repair: molecular and cellular mechanisms
J Invest Dermatol
(2007) - et al.
Harnessing growth factors to influence wound healing
Clin Plast Surg
(2012) - et al.
Differences in cellular infiltrate and extracellular matrix of chronic diabetic and venous ulcers versus acute wounds
J Invest Dermatol
(1998) - et al.
Dysregulation of monocyte/macrophage phenotype in wounds of diabetic mice
Cytokine
(2011) - et al.
Diabetic foot ulcers
Lancet
(2003) - et al.
A transgenic mouse model of inducible macrophage depletion: effects of diphtheria toxin-driven lysozyme M-specific cell lineage ablation on wound inflammatory, angiogenic, and contractive processes
Am J Pathol
(2009) - et al.
Selective and specific macrophage ablation is detrimental to wound healing in mice
Am J Pathol
(2009)
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