Deoxynivalenol Damages Corneal Epithelial Cells and Exacerbates Inflammatory Response in Fungal Keratitis.

BACKGROUND: Fungal keratitis (FK) is a kind of infectious keratopathy with a high rate of blindness worldwide. Deoxynivalenol (DON) has been proven to have multiple toxic effects on humans and animals. OBJECTIVES: The aim of this study was to explore a possible pathogenic role of DON in FK. METHODS: We first made an animal model of FK in New Zealand white rabbits, and then attempted to detect DON in a culture medium in which Fusarium solani had been grown and also in the corneal tissue of the animal model of Fusarium solani keratitis. Next, a model of DON damage in human corneal epithelial cells (HCECs) was constructed to evaluate effects of DON on the activity, migration ability, cell cycle, and apoptosis in the HCECs. Then, putative the toxic damaging effects of DON on rabbit corneal epithelial cells and the impact of the repair cycle were studied. The expression levels of inflammatory factors in the corneas of the animal model and in the model of DON-damaged HCECs were measured. RESULTS: The Fusarium solani strain used in this study appeared to have the potential to produce DON, since DON was detected in the corneal tissue of rabbits which had been inoculated with this Fusarium solani strain. DON was found to alter the morphology of HCECs, to reduce the activity and to inhibit the proliferation and migration of HCECs. DON also induced the apoptosis and S-phase arrest of HCECs. In addition, DON was found to damage rabbit corneal epithelial cells, to prolong the corneal epithelial regeneration cycle, and to be associated with the upregulated expression of inflammatory factors in HCECs and rabbit corneas. CONCLUSIONS: DON appears to have a toxic damaging effect on HCECs in FK, and to induce the expression of inflammatory factors, leading to the exacerbation of keratitis and the formation of new blood vessels. Future studies will explore the possibility of developing a test to detect DON in ophthalmic settings to aid the rapid diagnosis of FK, and to develop DON neutralizers and adsorbents which have the potential to improve keratocyte status, inhibit apoptosis, and alleviate inflammation, therein providing new thinking for therapy of clinical FK.

Quantitative assessment of retinal vessel density and thickness changes in internal carotid artery stenosis patients using optical coherence tomography angiography.

OBJECTIVE: To quantitatively assess the retinal features of patients with different degrees of internal carotid artery stenosis (ICAS), particularly mild ICAS patients, utilizing optical coherence tomography angiography (OCTA). METHODS: Thirty-two mild ICAS patients (mild ICAS group), 34 moderate to severe ICAS patients (nonmild ICAS group), and 40 controls were enrolled in this study. Retinal vessel density was quantitatively measured by OCTA, including radial peripapillary capillary vessel density (RPC-VD), superficial and deep capillary plexus vessel density (SCP/DCP-VD). Structural parameters were collected from optical coherence tomography (OCT), including retinal thickness and subfoveal choroidal thickness (SFCT). Furthermore, LASSO-penalized logistic regression was used to construct the diagnostic model based on retinal parameters. ROC curves and nomogram plots were used to assess the diagnostic ability of this model for ICAS. RESULTS: The macular SCP-VD of mild ICAS patients was significantly lower than that of controls and lower than that of nonmild ICAS patients (all p < 0.05). However, there was no difference among the three groups in terms of DCP-VD (p > 0.05). RPC-VD could effectively discriminate between the mild ICAS group and the nonmild ICAS group (p = 0.005). For structural OCT, only the SFCT decreased as the ICAS degree increased (p < 0.05). Diagnostic scores based on retinal parameters showed a strong diagnostic capability for mild ICAS (AUC = 0.8656). CONCLUSION: Mild ICAS patients exhibited distinct retinal features compared to nonmild ICAS patients and control subjects. OCTA potentially represents a promising method for the early detection of ICAS patients and the noninvasive surveillance of haemodynamic changes in those patients.

Development of a Novel Retina-Based Diagnostic Score for Early Detection of Major Depressive Disorder: An Interdisciplinary View.

Background: Clinically effective markers for the diagnosis of major depressive disorder (MDD) are lacking. Alterations in retinal features are closely related to the pathophysiological progression of MDD. However, the reliable retina-related diagnostic model for MDD remains to be developed. Thus, our study aimed to quantitatively evaluate retinal vascular and structural changes in MDD patients and to develop a reliable diagnostic model of MDD based on retinal parameters. Methods: Seventy-eight patients with MDD and 47 healthy controls (HCs) underwent retinal vessel density and structure examination using optical coherence tomography angiography and visual field examination using perimetry. Independent-sample t test was used to assess the differences in retinal parameters between the groups. Meanwhile, we constructed the corresponding retina-based diagnostic model by LASSO logistic regression. Finally, the diagnostic ability of the model was evaluated by area under the curve (AUC) of receiver operating characteristic curves and calibration plot of nomogram. Results: MDD patients showed lower retinal vessel density (including radial peripapillary capillary vessel density, superficial and deep capillary plexus vessel density), thinner subfoveal choroidal thickness, and poorer visual fields compared to HCs (all p < 0.05). Furthermore, a retina-based diagnostic model was constructed and shows a strong diagnostic capability for MDD (AUC = 0.9015, p < 0.001). Conclusion: Patients with MDD showed distinct retinal features compared to HCs. The retina-based diagnostic model is expected to be a necessary complement to the diagnosis of MDD.

Exosomes and autophagy in ocular surface and retinal diseases: new insights into pathophysiology and treatment.

BACKGROUND: Ocular surface and retinal diseases are widespread problems that cannot be ignored in today's society. However, existing prevention and treatment still have many shortcomings and limitations, and fail to effectively hinder the occurrence and development of them. MAIN BODY: The purpose of this review is to give a detailed description of the potential mechanism of exosomes and autophagy. The eukaryotic endomembrane system refers to a range of membrane-bound organelles in the cytoplasm that are interconnected structurally and functionally, which regionalize and functionalize the cytoplasm to meet the needs of cells under different conditions. Exosomal biogenesis and autophagy are two important components of this system and are connected by lysosomal pathways. Exosomes are extracellular vesicles that contain multiple signaling molecules produced by multivesicular bodies derived from endosomes. Autophagy includes lysosome-dependent degradation and recycling pathways of cells or organelles. Recent studies have revealed that there is a common molecular mechanism between exosomes and autophagy, which have been, respectively, confirmed to involve in ocular surface and retinal diseases. CONCLUSION: The relationship between exosomes and autophagy and is mostly focused on fundus diseases, while a deeper understanding of them will provide new directions for the pathological mechanism, diagnosis, and treatment of ocular surface and retinal diseases.

Exosomes From Human Umbilical Cord Mesenchymal Stem Cells Treat Corneal Injury via Autophagy Activation.

Corneal injury (CI) affects corneal integrity and transparency, deteriorating the patient's quality of life. This study aimed to explore the molecular mechanisms by which exosomes secreted from human umbilical cord mesenchymal stem cells (hucMSC-Exos) affect autophagy in human corneal epithelial cells (HCECs) and CI models. We isolated and identified hucMSC-Exos using nanoparticle tracking analysis, transmission electron microscopy, and western blotting. The effects of hucMSC-Exos combined with autophagy regulators on HCECs and CI mice were assessed using cell viability assays, scratch assay, cell cycle assay, apoptosis assay, corneal fluorescein staining, haze grades, pathological examinations, western blotting, and quantitative polymerase chain reaction (qPCR). In vitro results indicated that hucMSC-Exos combined with the autophagy activator had positive effects in promoting the cell proliferation, migration capacity, and the cell cycle by upregulating the proportions of cells in the S phase and the expression of PCNA, Cyclin A, Cyclin E, and CDK2. Meanwhile, the combination treatment reduced the apoptotic rate of HCECs. In vivo results indicated that hucMSC-Exos especially combined them with the autophagy activator significantly alleviated corneal epithelial defects and stromal opacity, reduced the levels of the apoptotic markers Bax and cleaved Caspase-3, reduced the inflammatory response products TNF-α, IL-1ß, IL-6, and CXCL-2, and increased the Bcl-2. This was achieved by upregulating pAMPK/AMPK and pULK1/ULK1 ratios, and Beclin-1 and LC3B II/I, and by downregulating the pmTOR/mTOR ratio and p62. In contrast, clinical indications, apoptosis, and inflammation were aggravated after the application of the autophagy inhibitor. HucMSC-Exos combined with an autophagy activator significantly enhanced HCECs functions and alleviated corneal defects, apoptosis, and inflammation by activating the autophagy signaling pathway, AMPK-mTOR-ULK1, providing a new biological therapy for corneal wound healing and ocular surface regeneration.

Self-assembling hydrogel loaded with 5-FU PLGA microspheres as a novel vitreous substitute for proliferative vitreoretinopathy.

The vitreous substitute for proliferative vitreoretinopathy (PVR) surgery remains an unmet clinical need in ophthalmology. In our study, we developed an in situ formed hydrogel by crosslinking polyvinyl alcohol (PVA) and chitosan as a potential vitreous substitute. 5-fluorouracil (5-FU) Poly (lactic-co-glycolic acid) (PLGA) microspheres were developed and loaded onto the PVA/chitosan hydrogels to treat PVR. In vitro, PVA/chitosan hydrogels at four concentrations were subjected to morphological, physical, rheological analyses, and cytotoxicity was evaluated together with the characterization of 5-FU PLGA microspheres. In vivo, pharmacologically induce PVR rabbits were performed a vitrectomy. In the PVA group, 3% PVA/chitosan hydrogel was injected into the vitreous cavity. In the PVA/MS group, 3% PVA/chitosan hydrogel and 5-FU PLGA microspheres were injected. In the Control group, phosphate-buffered saline was injected. Therapeutic efficacy was evaluated with postoperative examinations and histological analyses. This study demonstrated that the 3% PVA/chitosan hydrogel showed properties similar to those of the human vitreous and could be a novel in situ crosslinked vitreous substitute for PVR. Loading 5-FU PLGA microspheres onto this hydrogel may represent an effective strategy to improve the prognosis of PVR.

Corneal autophagy and ocular surface inflammation: A new perspective in dry eye.

Dry eye disease (DED), a multifactorial ocular surface disorder affecting millions of individuals worldwide, is characterized by inflammation and damage to the ocular surface. It is unclear whether corneal autophagy participates in ocular surface inflammation observed in DED. To test this involvement, dry eye (DE) was induced in female C57BL/6 mice housed in a controlled environment by subcutaneous injection of scopolamine. Expression of the autophagy-related proteins LC3B and ATG5 and activation of autophagy were detected in the corneas of these mice. Treatment with LYN-1604, an activator of autophagy, alleviated the clinical indications in DE mice, including tear production and corneal fluorescence staining. LYN-1604 also reduced the corneal levels of inflammatory response products, including tumor necrosis factor alpha (TNF-α) and matrix metalloproteinases-3 and -9. By contrast, treatment of DE mice with the autophagy inhibitor 3-MA, exacerbated the clinical indications of DE and increased the levels of inflammatory response products. This is the first study to show that autophagy could regulate the level of ocular surface inflammation, suggesting that agents that regulate autophagy could relieve ocular surface inflammation and treat DED.