Unraveling the molecular dynamics of wound healing: integrating spatially resolved lipidomics and temporally resolved proteomics

Understanding the spatial-temporal molecular dynamics of wound healing is crucial for devising effective treatments. Three-dimensional mass spectrometry imaging (3D MSI) enables the comprehensive visualization of molecular distribution throughout skin layers, offering valuable insights into the wound healing process. However, traditional 3D MSI often faces challenges in maintaining data integrity and accurate image registration in the third dimension. To address this, we employed infrared matrix-assisted laser desorption electrospray ionization (IR-MALDESI), a hybrid ambient ionization technique capable of sequential imaging through consecutive ablation events for precise 3D image reconstruction. Herein, 3D IR-MALDESI MSI was used to compare the lipidome of fresh-frozen wound samples at three stages of wound healing (inflammation, proliferation, and remodeling) with the healthy skin of SKH- 1 mice. Supplementing this data with a refined LC-MS-based proteomics protocol on selected wound biopsies, our integrated approach deepens our understanding of the molecular intricacies inherent in tissue regeneration.