Abstract
High mobility group box 1 (HMGB1) is tightly connected to the process of tissue organization upon tissue injury. Here we show that HMGB1 controls epithelium and connective tissue regeneration both in vivo and in vitro during palatal wound healing. Heterozygous HMGB1 (Hmgb1+/−) mice and Wild-type (WT) mice were subjected to palatal injury. Maxillary tissues were stained with Mallory Azan or immunostained with anti-HMGB1, anti-proliferating cell nuclear antigen (PCNA), anti-nuclear factor-κB (NF-κB) p50 and anti-vascular endothelial growth factor (VEGF) antibodies. Palatal gingival explants were cultured with recombinant HMGB1 (rHMGB1) co-treated with siRNA targeting receptor for advanced glycation end products (RAGEs) for cell migration and PCNA expression analysis. Measurement of the wound area showed differences between Hmgb1+/− and WT mice on Day 3 after wounding. Mallory Azan staining showed densely packed of collagen fibers in WT mice, whereas in Hmgb1+/− mice weave-like pattern of low density collagen bundles were present. At three and seven days post-surgery, PCNA, NF-κB p50 and VEGF positive keratinocytes of WT mice were greater than that of Hmgb1+/− mice. Knockdown of RAGE prevents the effect of rHMGB1-induced cell migration and PCNA expression in gingival cell cultures. The data suggest that HMGB1/RAGE axis has crucial roles in palatal wound healing.
Highlights
In tissues skin and oral mucosa tissues, wound healing encompasses a number of overlapping phases, including inflammation, tissue formation and tissue remodeling [1]
DNA extraction from mouse tail and genotyping Polymerase Chain Reaction (PCR) separated by agarose gel electrophoresis has been performed in Hmgb1+/− and Wild-type (WT) mice (Figure 1A)
Our data replicated previous findings that Hmgb1+/− mice contained approximately one-half the level of High mobility group box 1 (HMGB1) mRNA compared with WT mice [26]
Summary
In tissues skin and oral mucosa tissues, wound healing encompasses a number of overlapping phases, including inflammation, tissue formation and tissue remodeling [1]. Ligand binding results in RAGE-dependent sustained nuclear factor-kappa B (NF-κB) activation [24] as well as in wound healing promotion [25]. These reports indicate that HMGB1 is a multifunctional cytokine involved in inflammatory responses and tissue repair. Show for the first time, that HMGB1/RAGE axis has crucial roles in palatal wound healing, by regulating collagen accumulation, cell proliferation and cell migration. Taken together, these results bring scientific support to the possible application of HMGB1 in regenerative medicine
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