Bioactive Phenolics from Vinegar-Egg Accelerates Acute Wound Healing by Activation of Focal Adhesion and Mitogen-Activated Protein Kinase Signaling.

Abstract

<h4>Background/objectives</h4>Vinegar-egg is a traditional health-promoting beverage prepared by soaking eggs in vinegar. While both eggs and vinegar are common dietary components with well-documented nutritional and pharmacological activities, eggs treated with vinegar have been rarely studied. This study aims to identify and characterize bioactive compounds in vinegar-egg and investigate their potential wound-healing activities.<h4>Methods</h4>The vinegar-egg extract was analyzed using liquid chromatography-mass spectrometry (LC-MS) and column chromatography, including HPLC purification, which led to the isolation of four phenolic compounds.<h4>Results</h4>These compounds were identified as 4-hydroxybenzoic acid (<b>1</b>), vanillic acid (<b>2</b>), methyl syringate (<b>3</b>), and leptosperin (<b>4</b>) using ESI-MS, UV, and NMR spectroscopic data. Among the isolates, 4-hydroxybenzoic acid (<b>1</b>) and vanillic acid (<b>2</b>) demonstrated wound-healing properties in mouse embryonic fibroblast (MEF) cells. None of the compounds, 4-hydroxybenzoic acid (<b>1</b>), vanillic acid (<b>2</b>), methyl syringate (<b>3</b>), or leptosperin (<b>4</b>), exhibited cytotoxicity in PC12, AGS, MEF, or MDA-MB-231 cells. Notably, 4-hydroxybenzoic acid (<b>1</b>) enhanced cell motility by 2.59-fold and cell invasion by 1.20-fold, while vanillic acid (<b>2</b>) increased cell motility by 2.69-fold and cell invasion by 1.23-fold. Western blot analysis revealed that treatment with 4-hydroxybenzoic acid (<b>1</b>) and vanillic acid (<b>2</b>) increased the phosphorylation of focal adhesion kinase (p-FAK) and matrix metalloproteinase 2 (MMP-2). Furthermore, both compounds elevated the phosphorylation of p38, a key regulator in wound-healing pathways.<h4>Conclusions</h4>These findings demonstrate that 4-hydroxybenzoic acid (<b>1</b>) and vanillic acid (<b>2</b>) accelerate wound healing through the activation of focal adhesion and mitogen-activated protein kinase (MAPK) signaling pathways. These results highlight vinegar-egg as a promising therapeutic candidate for wound healing.