Molecular profiling and functional analyses of umbilical cord mesenchymal stem cell-derived extracellular vesicles for dry eye diseases.

Abstract

Aim: Dry eye disease (DED) is a multifactorial disorder characterized by tear film instability, ocular surface damage, and inflammation. This study investigates the molecular landscape and the therapeutic potential of umbilical cord mesenchymal stem cell-derived extracellular vesicles (UCMSC-EVs) as a standardized, cell-free treatment for DED. Methods: UCMSC-EVs were produced from UCMSCs in a scalable 3D bioreactor system and enriched via tangential flow filtration (TFF). The physicochemical properties of UCMSC-EVs were characterized. The molecular cargo of UCMSC-EVs was analyzed via proteomics analysis and microRNA (miRNA) microarray profiling across multiple production lots. Functional potency of UCMSC-EVs was assessed using a human corneal epithelial cell line (HCE-S) wound healing assay and two human inflammation models. Results: UCMSC-EVs exhibited a consistent cargo profile enriched with proteins and miRNAs targeting DED-related pathways, with high proteomic correlation (Pearson's r > 0.8) across independent lots. Functional assays demonstrated that UCMSC-EVs promoted HCE-S wound healing and suppressed pro-inflammatory cytokines in a dose-dependent manner. Furthermore, a significant negative correlation was identified between PEDF (Pigment Epithelium-Derived Factor) cargo concentration and interferon gamma (IFN-γ) secretion (Pearson's r = -0.724, P = 0.005), providing a quantitative link between molecular identity and anti-inflammatory potency. Conclusion: UCMSC-EVs possess favorable physicochemical and molecular characteristics, and demonstrate robust regenerative and anti-inflammatory properties in vitro. The high manufacturing consistency and the identification of key molecular drivers, such as PEDF, support the potential of UCMSC-EV as a well-characterized and viable cell-free therapy for mitigating DED progression.