Investigation of the Effect of Mesenchymal Stem Cells with Increased E-cadherin and ICAM1 Gene Expression Using the Crispra/dCas9 System on Wound Healing in Mouse Models

Cell adhesion molecules are proteins that bind to the surface of cells. These proteins play a vital role in tissue wound healing by binding to other cells or the extracellular matrix. Intracellular Adhesion Molecule-1 (ICAM-1) and Epithelial Cadherin (E-cadherin), which belong to the extracellular matrix family, play a key role in wound healing in skin tissue. Stem cells and CRISPR/Cas systems have been increasingly utilized in the treatment of dermal wounds in recent years, within the scope of tissue engineering and regenerative medicine. In this study, adipose tissue stem cells (ADSCs) were obtained from mice, and their characterization was investigated using inverted phase-contrast microscopy, and flow cytometry. The ICAM-1 and/or E-cadherin genes in these cells were overexpressed using the lentiviral-mediated CRISPRa/dCas9 system. The effects of overexpression of these genes on gene expression, viability, adhesion, proliferation, and migration of ADSCs were investigated in vitro using inverted fluorescence phase-contrast microscopy, RT-qPCR, and flow cytometry. Furthermore, genetically modified ADSCs were transplanted into full-thickness wound defects in vivo, and their effects on wound healing were examined both morphologically and histologically. In conclusion, ICAM-1 and/or E-cadherin genes were successfully overexpressed in ADSCs using the CRISPRa/dCas9 system. Overexpression of these genes in ADSCs resulted in increased surface adhesion times, as well as enhanced proliferation and migration in the wound assay. In addition to the in vitro findings on the wound-healing effects of ADSCs, in vivo animal studies have also confirmed that ADSCs significantly reduce wound size, accelerate reepithelialization from wound edges, and initiate vascularization at earlier stages of wound healing. Taken together, these findings suggest that overexpressing ICAM-1 and/or E-cadherin genes in ADSCs via the CRISPRa/dCas9 system significantly enhanced the adhesion properties of these cells, resulting in increased binding to the wound site and significantly support dermal wound healing.