Highly Diluted Acetylcholine Promotes Wound Repair in an In Vivo Model

• Published in: Advances in wound care (New Rochelle, N.Y.) Vol. 7; no. 4; p. 121
• Main Authors: Uberti, Francesca, Morsanuto, Vera, Ghirlanda, Sabrina, Ruga, Sara, Clemente, Nausicaa, Boieri, Cristina, Boldorini, Renzo, Molinari, Claudio
• Format: Journal Article
• Language: English
• Published: United States 01.04.2018
• Subjects: wound healing; acetylcholine; low dose; sequential kinetic activation
• ISSN: 2162-1918
• 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.