Clinical observation of removal of the necrotic corneal tissue combined with conjunctival flap covering surgery under the guidance of the AS-OCT in treatment of fungal keratitis

AIM: To study the clinical observation of removal of the necrotic corneal tissue combined with conjunctival flap covering surgery under the guidance of the AS-OCT in treatment of fungal keratitis. METHODS:A retrospective study was done to 10 patients (10 eyes) who had accepted removal of the necrotic corneal tissue combined with conjunctival flap covering surgery for fungal keratitis,the diagnosis by corneal scraping and smear examination or confocal microscopy check hyphae. Local and systemic antifungal therapy more than one week for all patients, corneal ulcer enlarge or no shrink. Slit lamp microscope examination the diameter of corneal ulcer about 2mm-4mm. Anterior segment optical coherence tomography (AS-OCT)examine the depth of corneal ulcer between 1/3-1/2, infiltrate corneal stroma about 20um-80um,the diameter of corneal ulcer about 3mm-6mm.Type-B ultrasonic exclusion endophthalmitis. Complete removal lesions until transparent of stoma, make conjunctival flap equal or greater than ulcer 1mm nearby conjunctiva. Continued antifungal therapy. The vision, fungal recurrence, conjunctival flap rollback or desquamate were analysed. ' RESULTS:Ten patients had success done this surgery, the corneal ulcer was not enlarge and healing afteroperation. 7 cases were bridging conjunctival flap and 3cases were single conjunctival flap. Preoperation vision above 0.1 had 8 cases,7 cases had vision above 0.1 one week after surgery, while 1 cases vision droped from 0.3 to 0.05.There was not recurrent for fungal,2 cases conjunctival flap rollback:1 case was bridging and 1case was single flap, no conjunctival flap desquamate. CONCLUSION: It is safe and effective to perform removal of the necrotic corneal tissue combined with conjunctival flap covering surgery under the guidance of the AS-OCT in treatment of fungal keratitis which werenot sensitive or aggravate for antifungal drugs.

Acellular porcine corneal matrix as a carrier scaffold for cultivating human corneal epithelial cells and fibroblasts in vitro

AIM：To investigate the feasibility of corneal anterior lamellar reconstruction with human corneal epithelial cells and fibroblasts,and an acellular porcine cornea matrix（APCM） in vitro.·METHODS：The scaffold was prepared from fresh porcine corneas which were treated with 0.5%sodium dodecyl sulfate（SDS）solution and the complete removal of corneal cells was confirmed by hematoxylin-eosin（HE）staining and 4’,6-diamidino-2-phenylindole（DAPI）staining.Human corneal fibroblasts and epithelial cells were cultured with leaching liquid extracted from APCM,and then cell proliferative ability was evaluated by MTT assay.To construct a human corneal anterior lamellar replacement,corneal fibroblasts were injected into the APCM and cultured for 3d,followed by culturing corneal epithelial cells on the stroma construction surface for another 10d.The corneal replacement was analyzed by HE staining,and immunofluorescence staining.·R ESULTS：Histological examination indicated that there were no cells in the APCM by HE staining,and DAPI staining did not detect any residual DNA.The leaching liquid from APCM had little influence on the proliferation ability of human corneal fibroblasts and epithelial cells.At 10d,a continuous 3 to 5 layers of human corneal epithelial cells covering the surface of the APCM was observed,and the injected corneal fibroblasts distributed within the scaffold.The phenotype of the construction was similar to normal human corneas,with high expression of cytokeratin 12 in the epithelial cell layer and high expression of Vimentin in the stroma.·CONCLUSION：Corneal anterior lamellar replacement can be reconstructed in vitro by cultivating human corneal epithelial cells and fibroblasts with an acellular porcine cornea matrix.This laid the foundation for the further transplantation in vitro.

A highly selective fluorescent probe for visualizing dry eye disease-associated viscosity variations

Dry eye disease(DED) is a multifactorial chronic inflammatory disease of the ocular surface with complex and unclear etiology. The development of reliable detection tools for the pathology of DED will benefit its treatment, but it is still lacking. In parallel, it has been discovered recently that viscosity changes are involved in inflammation processes. In this regard, we constructed a fluorescent probe V5with an asymmetric donor-acceptor-donor(D-A-D) feature after rational structural modulation for viscosity detection during DED progression. The probe manifested a remarkable fluorescence enhancement(110 folds) in highly viscous conditions without interferences from polarity and reactive species. Specifically, no aggregation effect of the probe was found in glycerol. Moreover, viscosity increment in human corneal epithelial cells(HCECs) induced by hyperosmosis and inflammation was monitored, and ferroptosis in HCECs also led to the viscosity elevation. A reactive oxygen species(ROS)-dependent viscosity changes during DED progression is demonstrated. Finally, viscosity change in corneal epithelial cell layer from mice treated by scopolamine was also visualized for the first time. We anticipate this work can provide a new lens to the pathogenesis study and diagnosis of DED and other ophthalmic diseases using fluorescence methods.