Uberti Francesca, Morsanuto Vera, Ghirlanda Sabrina, Ruga Sara, Clemente Nausicaa, Boieri Cristina, Boldorini Renzo, Molinari Claudio
Physiology Laboratory, Department of Translational Medicine, UPO, Novara, Italy.
Immunology Laboratory, Department of Health Sciences, UPO, Novara, Italy.
Adv Wound Care (New Rochelle). 2018 Apr 1;7(4):121-133. doi: 10.1089/wound.2017.0766.
Wound healing is a dynamic, interactive, and complex process that involves a series of events, including inflammation, migration, proliferation, granulation tissue formation, and matrix remodeling. Despite the high frequency of serious slow-healing wounds, there is still no adequate therapy. The aim of this study is to evaluate a new highly diluted acetylcholine (Ach) formulation obtained through a sequential kinetic activation (SKA) method applied to a wound healing model to verify the hypothesis that a low dose of Ach could be a more physiological stimulus for healing, by stimulating muscarinic and nicotinic receptors and their related intracellular pathways. Two different concentrations (10 fg/mL and 1 pg/mL) and two formulations (either kinetically or nonkinetically activated) of Ach were used to verify the wound healing process. Area closure, histological aspect, and nicotinic and muscarinic receptors, matrix metalloproteinase 9 (MMP-9), Nestin, and von Willebrand's factor have been assessed by Western blot or ELISA and compared to 147 ng/mL Ach, used as positive control. Moreover, the systemic effect through plasmatic radical oxygen species (ROS) production and Ach concentration has been evaluated. Ach SKA 1 pg/mL revealed a significant capacity to restore the integrity of tissue compared to other formulation and this effect was more evident after a single administration. Topical application on skin of Ach SKA 1 pg/mL accelerates wound closure stimulating non-neuronal cholinergic system. Our results demonstrate for the first time the importance in an model of highly diluted SKA Ach during wound healing, suggesting a potential use in skin disease.
伤口愈合是一个动态、相互作用且复杂的过程，涉及一系列事件，包括炎症、迁移、增殖、肉芽组织形成和基质重塑。尽管严重的愈合缓慢伤口发生率很高，但仍然没有足够的治疗方法。本研究的目的是评估一种通过顺序动力学激活（SKA）方法获得的新型高稀释乙酰胆碱（Ach）制剂，将其应用于伤口愈合模型，以验证低剂量Ach通过刺激毒蕈碱和烟碱受体及其相关细胞内途径可能是更生理性的愈合刺激因素这一假设。使用两种不同浓度（10 fg/mL和1 pg/mL）以及两种制剂（动力学激活或非动力学激活）的Ach来验证伤口愈合过程。通过蛋白质印迹法或酶联免疫吸附测定法评估了伤口面积闭合、组织学特征、烟碱和毒蕈碱受体、基质金属蛋白酶9（MMP-9）、巢蛋白和血管性血友病因子，并与用作阳性对照的147 ng/mL Ach进行了比较。此外，还评估了通过血浆活性氧（ROS）产生和Ach浓度产生的全身效应。与其他制剂相比，1 pg/mL的Ach SKA显示出显著的恢复组织完整性的能力，且这种效应在单次给药后更为明显。1 pg/mL的Ach SKA局部应用于皮肤可通过刺激非神经元胆碱能系统加速伤口闭合。我们的结果首次证明了在伤口愈合过程中高稀释SKA Ach模型的重要性，表明其在皮肤病中的潜在应用价值。
