NK cells are best known for their killing of virus-infected cells and tumor cells via release of cytotoxic factors. However, NK cells can also produce growth factors and cytokines, and thus have the potential to influence physiological processes such as wound healing. In this study, we test the hypothesis that NK cells play a physiological role in skin wound healing of C57BL/6J mice. Immunohistochemical and flow cytometry assays showed that NK cells accumulate in excisional skin wounds, peaking on day 5 postinjury. We also found that NK cells proliferate locally in wounds, and blocking IL-15 activity locally reduces NK cell proliferation and accumulation in wounds. Wound NK cells exhibit primarily a mature CD11b+CD27− and NKG2A+NKG2D− phenotype and express LY49I and proinflammatory cytokines such as IFN-γ, Tnf-a, and Il-1β. Systemic depletion of NK cells resulted in enhanced re-epithelization and collagen deposition, suggesting a negative role for these cells in skin wound healing. Depletion of NK cells did not influence accumulation of neutrophils or monocytes/macrophages in wounds but did reduce expression of IFN-γ, Tnf-a, and Il-1β, indicating that NK cells contribute to proinflammatory cytokine expression in wounds. In short, NK cells may impede physiological wound healing via production of proinflammatory cytokines.

Significance: Chronic skin wounds are a significant health problem around the world, often leading to amputation and even death. Although persistent inflammation is a hallmark of these poorly healing wounds, few available therapies have been designed to target inflammation. In this review, we summarize available evidence of the role of the NOD-like receptor pyrin domain containing 3 (NLRP3) inflammasome in impaired wound healing and describe strategies to inhibit the inflammasome to improve wound healing. Recent Advances: The NLRP3 inflammasome plays an important physiological role in skin wound healing, during which transient inflammasome activity contributes to both epidermal and dermal healing. In contrast, sustained activity of the NLRP3 inflammasome leads to impaired epidermal and dermal healing associated with diabetes. Of importance, preclinical studies have demonstrated that inhibiting the NLRP3 inflammasome-induced resolution of inflammation, increased granulation tissue formation and collagen deposition, and accelerated reepithelialization and wound closure. Critical Issues: NLRP3 inflammasome inhibitors have appealing potential for translation into therapies for chronic wounds. Although preclinical studies have shown promising results, there is a need for human/clinical studies to evaluate dosing formulations, potential therapeutic effects, dose–response relationships, and possible side effects. Future Directions: Among strategies to inhibit the NLRP3 inflammasome, glyburide, metformin, peroxisome proliferator-activated receptor agonists, and the dipeptidyl peptidase 4 inhibitor saxagliptin appear to be closest to clinical translation, as these drugs are already Food and Drug Administration approved for other indications. Future clinical studies are needed to develop topical formulations of these drugs, and to assess the safety and efficacy of these inhibitors, to improve healing of chronic wounds.

Objective: Chronic wounds associated with diabetes are an important public health problem demanding new treatments to improve wound healing and decrease amputations. Monocytes/macrophages play a key role in sustained inflammation associated with impaired healing and local administration of peroxisome proliferator-activated receptor (PPAR)γ agonists may modulate macrophage, improving healing. In this study, we investigated the effects of GQ-11, a partial/dual PPARα/γ agonist, on macrophage function and wound healing in diabetes. Approach: Wounds were surgically induced at the dorsum of C57BL/6J and BKS.Cg-Dock7^m +/+ Lepr^db/J (db/db) mice and treated with hydrogel (vehicle), pioglitazone or GQ-11, for 7 or 10 days, respectively. After treatment, wounds were analyzed histologically and by quantitative PCR (qPCR). In addition, bone marrow-derived macrophages (BMDM) were cultured from C57BL/6J mice and treated with vehicle, pioglitazone, or GQ-11, after challenge with lipopolysaccharide or interleukin-4 to be analyzed by qPCR and flow cytometry. Results: GQ-11 treatment upregulated anti-inflammatory/pro-healing factors and downregulated pro-inflammatory factors both in wounds of db/db mice and in BMDM. Innovation: Wounds of db/db mice treated with GQ-11 exhibited faster wound closure and re-epithelization, increased collagen deposition, and less Mac-3 staining compared with vehicle, providing a new approach to treatment of diabetic wound healing to prevent complications. Conclusion: GQ-11 improves wound healing in db/db mice, regulating the expression of pro- and anti-inflammatory cytokines and wound growth factors, leading to increased re-epithelization and collagen deposition.

Insulin-like growth factor (IGF)-1 plays important role in tissue repair through its ability to stimulate wound cell activity. While IGF-1 is expressed locally by wound cells, liver-derived IGF-1 is also present at high levels in the circulation, and the contributions of local vs circulating IGF-1 to wound levels remain undefined. The hypothesis of this study was that liver is a primary source of IGF-1 during skin wound healing. To test this hypothesis, we utilized a model that allows inducible ablation of IGF-1 specifically in liver of adult mice. We demonstrate that ablation of liver IGF-1 leads to >85% loss of circulating IGF-1 and ~60% decrease in wound IGF-1 during the proliferative phase of healing in both male and female mice. This reduction of liver-derived IGF-1 did not alter local mRNA expression of Igf1 in wounds. Knockdown of liver IGF-1 significantly delayed wound re-epithelialization and reduced granulation tissue formation and collagen deposition. Knockdown of liver IGF-1 also significantly reduced angiogenesis and resulted in persistent macrophage accumulation. In summary, liver is a primary source of IGF-1 in skin wounds and contributes to many aspects of both epithelial and dermal healing.