Nano-Bio Regenerative Medical Institute, College of Medicine, Hallym University, Chuncheon 200-702, South Korea.
Department of Otolaryngology-Head and Neck Surgery, Ilsong Memorial Institute of Head and Neck Cancer, Hallym University College of Medicine, 150 Seongan-ro, Gangdong-gu, Seoul, South Korea.
Acta Biomater. 2018 Feb;67:183-195. doi: 10.1016/j.actbio.2017.12.006. Epub 2017 Dec 11.
Silk fibroin (SF) is a well-studied biomaterial for tissue engineering applications including wound healing. However, the signaling mechanisms underlying the impact of SF on this phenomenon have not been determined. In this study, through microarray analysis, regulatory genes of NF-ĸB signaling were activated in SF-treated NIH3T3 cells along with other genes. Immunoblot analysis confirmed the activation of the NF-ĸB signaling pathway as SF induced protein expression levels of IKKα, IKKβ, p65, and the degradation of IκBα. The treatment of NIH3T3 cells with SF also increased the expression of cyclin D1, vimentin, fibronectin, and vascular endothelial growth factor (VEGF). The expression of these factors by SF treatment was abrogated when NF-ĸB was inhibited by a pharmacological inhibitor Bay 11-7082. Knockdown of NF-ĸB using siRNA of IKKα and IKKβ also inhibited the SF-induced wound healing response of the NIH3T3 cells in a wound scratch assay. Collectively, these results indicated that SF-induced wound healing through the canonical NF-κB signaling pathway via regulation of the expression of cyclin D1, vimentin, fibronectin, and VEGF by NIH3T3 cells. Using an in vivo study with a partial-thickness excision wound in rats we demonstrated that SF-induced wound healing via NF-κB regulated proteins including cyclin D1, fibronectin, and VEGF. The in vitro and in vivo data suggested that SF induced wound healing via modulation of NF-ĸB signaling regulated proteins.
Silk fibroin has been effectively used as a dressing for wound treatment for more than a century. However, mechanistic insight into the basis for wound healing via silk fibroin has not been elucidated. Here we report a key mechanism involved in silk fibroin induced wound healing both in vitro and in vivo. Using genetic- and protein-level analyses, NF-κB signaling was found to regulate silk fibroin-induced wound healing by modulating target proteins. Thus, the NF-κB signaling pathway may be utilized as a therapeutic target during the formulation of silk fibroin-based biomaterials for wound healing and tissue engineering.
丝素蛋白（SF）是一种经过充分研究的生物材料，可用于组织工程应用，包括伤口愈合。然而，SF 对这种现象的影响的信号机制尚未确定。在这项研究中，通过微阵列分析，在 SF 处理的 NIH3T3 细胞中激活了 NF-ĸB 信号的调节基因，以及其他基因。免疫印迹分析证实 NF-ĸB 信号通路被激活，因为 SF 诱导了 IKKα、IKKβ、p65 的蛋白表达水平和 IκBα 的降解。SF 处理 NIH3T3 细胞还增加了 cyclin D1、波形蛋白、纤连蛋白和血管内皮生长因子（VEGF）的表达。当使用药理学抑制剂 Bay 11-7082 抑制 NF-ĸB 时，SF 处理对这些因子的表达被阻断。使用 IKKα 和 IKKβ 的 siRNA 敲低 NF-ĸB 也抑制了 NIH3T3 细胞在划痕实验中的 SF 诱导的伤口愈合反应。总之，这些结果表明 SF 通过调节 NIH3T3 细胞中 cyclin D1、波形蛋白、纤连蛋白和 VEGF 的表达，通过经典的 NF-κB 信号通路诱导伤口愈合。通过大鼠部分厚度切除伤口的体内研究，我们证明了 SF 通过 NF-κB 调节的蛋白，包括 cyclin D1、纤连蛋白和 VEGF，诱导伤口愈合。体外和体内数据表明，SF 通过调节 NF-ĸB 信号调节蛋白诱导伤口愈合。
丝素蛋白已被有效地用作一个多世纪以来治疗伤口的敷料。然而，丝素蛋白通过伤口愈合的机制的深入了解尚未阐明。在这里，我们报告了一个关键的机制，涉及丝素蛋白诱导的体外和体内伤口愈合。使用遗传和蛋白质水平分析，发现 NF-κB 信号通过调节靶蛋白来调节丝素蛋白诱导的伤口愈合。因此，NF-κB 信号通路可作为基于丝素蛋白的生物材料用于伤口愈合和组织工程的配方中的治疗靶点。
