M2a Macrophage-Dominant Microenvironment in Inflammation Attenuation and Wound Healing of Human Corneal Endothelial Cells

초록

Purpose : To investigate the anti-inflammatory and wound-healing effects of an M2a macrophage-dominant microenvironment on human corneal endothelial cells (HCECs) in vitro. Methods : Two in vitro corneal endothelial inflammation models were developed: a lipopolysaccharide (LPS)-induced inflammation-only model and a dual inflammation-and-wound model. HCECs were co-cultured with M2a macrophages or treated with M1 macrophage-derived exosomes (M1 exo), M2a macrophage-derived exosomes (M2a exo) or epidermal growth factor (EGF)-preconditioned M2a-derived exosomes (EGF-M2a exo). Gene expression of inflammatory markers was assessed and HCEC proliferation was evaluated using cell growth curves. Results : Scratch wounding alone did not induce significant inflammation in HCECs, while LPS stimulation combined with scratch wounding markedly increased inflammatory responses. In the inflammation-only model, co-culture with M2a macrophages and M2a exo treatment significantly suppressed LPS-induced upregulation of IL6, IL1B, and ICAM1 in HCECs. In the dual inflammation-and-wound model, M2a exo showed limited efficacy, but EGF-M2a exo significantly reduced inflammatory marker expression. Cell growth analysis revealed that M2a exo promoted faster HCEC proliferation compared to M1 exo under non-inflammatory conditions, despite no significant differences in cell cycle-related genes. In LPS-stimulated HCECs, EGF-M2a exo treatment restored proliferation to levels comparable with non-inflammatory controls by day 5. Conclusions : An M2a macrophage-dominant microenvironment demonstrates anti-inflammatory and regenerative effects on inflamed HCECs. EGF preconditioning enhances these properties, suggesting a potential therapeutic approach for managing corneal endothelial inflammation and injury.