Anti-apoptotic and autophagic effect: Using conditioned medium from human bone marrow mesenchymal stem cells to treat human trabecular meshwork cells

Yu-Fan Chang; Yung-Hsin Cheng; Yu-Chieh Ko; Shih-Hwa Chiou; Catherine Jui-Ling Liu

doi:10.1016/j.reth.2022.12.002

Anti-apoptotic and autophagic effect: Using conditioned medium from human bone marrow mesenchymal stem cells to treat human trabecular meshwork cells

Regen Ther. 2022 Dec 23:22:50-58. doi: 10.1016/j.reth.2022.12.002. eCollection 2023 Mar.

Authors

Yu-Fan Chang^{1

2

3}, Yung-Hsin Cheng⁴, Yu-Chieh Ko^{2

3}, Shih-Hwa Chiou^{1

2

3}, Catherine Jui-Ling Liu^{2

3}

Affiliations

¹ Institute of Clinical Medicine, National Yang Ming Chiao Tung University, Taipei, 112, Taiwan.
² National Yang Ming Chiao Tung University School of Medicine, Taipei, Taiwan.
³ Department of Ophthalmology, Taipei Veterans General Hospital, Taipei, Taiwan.
⁴ Department of Materials Science and Engineering, National Taiwan University of Science and Technology, Taipei, Taiwan.

Abstract

Introduction: Glaucoma is a vision-threatening disease associated with accelerated aging of trabecular meshwork (TM) which results in elevated intraocular pressure (IOP). Increased oxidative stress in TM plays an important role in cellular molecular damage which leads to senescence. Autophagy is an intracellular lysosomal degradation process which is activated when cells are under stressful condition, and emerging studies have demonstrated increased expression of modulators of apoptosis and expression of autophagic cascade in ex-vivo TM specimens or cultured TM cells under oxidative stress. Recently, studies have shown neuroprotective and IOP-lowering effects after transplanting mesenchymal stem cells (MSCs) or injecting condition medium (CM) of MSCs into ocular hypertension animal models. However, knowledge of the underlying mechanism accounting for these effects is limited. Using condition medium (CM) from human bone marrow-derived mesenchymal stem cells (BM-MSCs), we investigated the effects of the CM derived from BM-MSCs on TM autophagy and apoptosis.

Methods: H₂O₂ was added to culture medium of human TM cells to mimic oxidative damage in glaucomatous eyes, and the autophagic and anti-apoptotic effects of BM-MSCs-derived CM was explored on the oxidatively damaged cells. Mitochondrial ROS production was examined by MitoSOX™, apoptosis was evaluated using terminal deoxynucleotidyl transferase (dUTP) nick end labeling (TUNEL) staining, and the expression of proteins involved in autophagy as well as extracellular matrix was investigated via Western blot.

Results: There were no significant differences in TM cell viability when the cells were treated with different concentrations of CM in the absence of oxidative stress. Cell viability was significantly higher in oxidatively damaged TM cells treated with 1X or 5X CM compared to untreated TM cells under oxidative stress. The mitochondrial ROS level significantly increased with oxidative stress, which was mitigated in the CM treatment groups. DNA fragmentation significantly decreased in oxidatively stressed TM cells after treatment with CM. LCB3 II/LCB3 I was significantly elevated in the oxidative stress group compared to the control group and was significantly decreased in the CM treatment groups. Expression of fibronectin was not significantly different among the groups.

Conclusion: The CM derived from human BM-MSCs has the capacity to rescue oxidatively damaged human TM cells associated with decreased autophagy and apoptosis. The BM-MSCs CM has potential for slowing down age- and disease-related degeneration of TM in patients with glaucoma, facilitating success in the control of IOP.

Keywords: Apoptosis; Autophagy; Mesenchymal stem cell; Trabecular meshwork cell.