Receptor-Interacting Protein Kinase 3 Deficiency Delays Cutaneous Wound Healing
Andrew Godwin1, Archna Sharma2, Weng-Lang Yang3
1Department of Surgery, Hofstra North Shore-LIJ School of Medicine, Manhasset, New York, United States of America.
Abstract:
Wound healing consists of a complex, dynamic and overlapping process involving inflammation, proliferation and tissue remodeling. A better understanding of wound healing process at the molecular level is needed for the development of novel therapeutic strategies. Receptor-interacting protein kinase 3 (RIPK3) controls programmed necrosis in response to TNF-α during inflammation and has been shown to be highly induced during cutaneous wound repair. However, its role in wound healing remains to be demonstrated. To study this, we created dorsal cutaneous wounds on male wild-type (WT) and RIPK3-deficient (Ripk3-/-) mice. Wound area was measured daily until day 14 post-wound and skin tissues were collected from wound sites at various days for analysis. The wound healing rate in Ripk3-/- mice was slower than the WT mice over the 14-day course; especially, at day 7, the wound size in Ripk3-/- mice was 53% larger than that of WT mice. H&E and Masson-Trichrome staining analysis showed impaired quality of wound closure in Ripk3-/- wounds with delayed re-epithelialization and angiogenesis and defected granulation tissue formation and collagen deposition compared to WT. The neutrophil infiltration pattern was altered in Ripk3-/- wounds with less neutrophils at day 1 and more neutrophils at day 3. This altered pattern was also reflected in the differential expression of IL-6, KC, IL-1β and TNF-α between WT and Ripk3-/- wounds. MMP-9 protein expression was decreased with increased Timp-1 mRNA in the Ripk3-/- wounds compared to WT. The microvascular density along with the intensity and timing of induction of proangiogenic growth factors VEGF and TGF-β1 were also decreased or delayed in the Ripk3-/- wounds. Furthermore, mouse embryonic fibroblasts (MEFs) from Ripk3-/- mice migrated less towards chemoattractants TGF-β1 and PDGF than MEFs from WT mice. These results clearly demonstrate that RIPK3 is an essential molecule to maintain the temporal manner of the normal progression of wound closure.
Insights
Receptor-interacting protein kinase 3 (RIPK3) is crucial for normal wound healing. RIPK3 deficiency impairs skin repair by delaying re-epithelialization, angiogenesis, and collagen deposition, highlighting its essential role in wound closure.
Area of Science:
- Molecular biology
- Immunology
- Dermatology
Background:
- Wound healing is a complex biological process involving inflammation, proliferation, and remodeling.
- Understanding molecular mechanisms is key to developing new therapeutic strategies for wound repair.
- Receptor-interacting protein kinase 3 (RIPK3) is involved in programmed necrosis and inflammation, and is upregulated during skin wound repair.
Purpose of the Study:
- To investigate the role of Receptor-interacting protein kinase 3 (RIPK3) in cutaneous wound healing.
- To determine if RIPK3 deficiency impacts the rate and quality of wound closure.
Main Methods:
- Dorsal cutaneous wounds were created in wild-type (WT) and RIPK3-deficient (Ripk3-/-) mice.
- Wound area was measured daily for 14 days.
- Skin tissues were analyzed using histology (H&E, Masson-Trichrome), and molecular techniques to assess inflammation, angiogenesis, and cell migration.
Main Results:
- Ripk3-/- mice exhibited slower wound healing rates compared to WT mice, with significantly larger wound sizes at day 7.
- Histological analysis revealed impaired re-epithelialization, angiogenesis, granulation tissue formation, and collagen deposition in Ripk3-/- wounds.
- Neutrophil infiltration patterns were altered, and the expression of inflammatory cytokines (IL-6, KC, IL-1β, TNF-α) and matrix metalloproteinases was dysregulated in Ripk3-/- wounds.
- Reduced microvascular density and delayed induction of proangiogenic factors (VEGF, TGF-β1) were observed in Ripk3-/- wounds.
- Mouse embryonic fibroblasts from Ripk3-/- mice showed decreased migration.
Conclusions:
- Receptor-interacting protein kinase 3 (RIPK3) plays an essential role in regulating the normal progression of cutaneous wound healing.
- RIPK3 is critical for proper inflammation resolution, angiogenesis, and tissue remodeling during the wound repair process.
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