Exosomes Derived from Mouse Adipose-Derived Mesenchymal Stem Cells Alleviate Benzalkonium Chloride-Induced Mouse Dry Eye Model via Inhibiting NLRP3 Inflammasome

• Published in: Ophthalmic research Vol. 65; no. 1; pp. 40 - 51
• Main Authors: Wang, Guifang, Li, Honghui, Long, Hongmei, Gong, Xileyuan, Hu, Shufang, Gong, Can
• Format: Journal Article
• Language: English
• Published: Basel, Switzerland S. Karger AG 01.02.2022
• Subjects: Animals; Benzalkonium Compounds - toxicity; Biological response modifiers; Dry Eye Syndromes - metabolism; Exosomes - metabolism; Fluorescein; Inflammasomes - adverse effects; Inflammasomes - metabolism; Interferon; Interleukins; Mesenchymal Stem Cells - metabolism; Mice; NLR Family, Pyrin Domain-Containing 3 Protein; Research Article; Stem cells; Surface active agents; China; Inflammation; Ocular surface; Mouse adipose-derived mesenchymal stem cell-derived exosomes; Dry eye; NLR family pyrin domain-containing 3; NLRP3 inflammasome
• DOI: 10.1159/000519458
• ISSN: 0030-3747; 1423-0259

Purpose: The objective of the study was to investigate efficacy and mechanisms of mouse adipose-derived mesenchymal stem cell-derived exosomes (mADSC-Exos) in the benzalkonium chloride (BAC)-induced mouse dry eye model. Methods: Exosomes in the mADSC culture supernatant were isolated by ultracentrifugation. Western blotting, nanoparticle tracking analysis, and transmission electron microscopy were used to characterize mADSC-Exos. An experimental mouse model of dry eye was established by instillation of 0.2% BAC. mADSC-Exos were administered following BAC treatment. The positive control group was treated with commercial eye drops (0.1% pranoprofen). Corneal fluorescein staining, tear secretion, and tear film break-up time (BUT) were evaluated, and histologic analysis of the cornea and conjunctiva was performed by hematoxylin and eosin and periodic acid-Schiff staining. Apoptosis in the corneal epithelium was detected with the terminal deoxynucleotidyl transferase dUTP nick-end labeling assay and by Western blotting. Levels of pro-inflammatory cytokines in the cornea and conjunctiva were evaluated by flow cytometry, and mRNA and protein levels of NLR family pyrin domain-containing 3 (NLRP3) pathway components were assessed by quantitative real-time PCR and Western blotting, respectively. Results: mADSC-Exos were characterized as vesicles with a bilayer membrane. The particle size distribution peak was at 134 nm. mADSC-Exos specifically expressed cluster of differentiation (CD)9, CD63, and CD81. mADSC-Exos treatment repaired ocular surface damage. Additionally, mADSC-Exos inhibited cell apoptosis, decreased the levels of interleukin (IL)-1β, IL-6, IL-1α, interferon (IFN)-γ, and tumor necrosis factor (TNF)-α, and increased levels of the anti-inflammatory cytokine IL-10. Meanwhile, NLRP3 inflammasome activation and upregulation of caspase-1, IL-1β, and IL-18 were reversed by mADSC-Exos. Conclusions: mADSC-Exos alleviate ocular surface inflammation, suggesting that it is a promising treatment for dry eye.