Three kinds of corneal host cells contribute differently to corneal neovascularization
Authors: Yifan Liu
Hua Qian
Fanqian Song
Yan Li
Haiyang Yu
Wentong Jiang
Yu Yan
Xiaoguang Li
Liyao Sun
Hongyan Ge
Chao Wang
Jing Cui
Xianling Tang
Xi Wei
Ping Liu
Journal: EBioMedicine (2019)
Discipline: CLINICAL MEDICINE
Keywords: CRYAA; Corneal cells; Galectin-8; MMP-2; MMP-9
Affiliations: Eye Hospital, First Affiliated Hospital, Harbin Medical University, 23 Youzheng Street, Harbin 150001, China; Department of Pharmacology, College of Pharmacy, Harbin Medical University and Heilongjiang Academy of Medical Sciences, 157 Baojian Road, Harbin 150081, China., Eye Hospital, First Affiliated Hospital, Harbin Medical University, 23 Youzheng Street, Harbin 150001, China., Center for Endemic Disease Control, Chinese Center for Disease Control and Prevention, Harbin Medical University, Harbin 150081, Heilongjiang Province, China., Department of Pharmacology, College of Pharmacy, Harbin Medical University and Heilongjiang Academy of Medical Sciences, 157 Baojian Road, Harbin 150081, China., Eye Hospital, First Affiliated Hospital, Harbin Medical University, 23 Youzheng Street, Harbin 150001, China. Electronic address: .

Abstract:  Background: Corneal neovascularization (angiogenesis and lymphangiogenesis) compromises corneal transparency and transplant survival, however, the molecular mechanisms of corneal host epithelial and stromal cells in neovascularization have not yet been fully elucidated. Furthermore, the contribution and mechanism of corneal host endothelial cells involved in neovascularization are largely unexplored. Methods: Liquid chromatography-mass spectrometry, immunoblotting, and ELISA were used to screen and identify potential neovascularization-related factors in human full-thickness vascularized corneal tissues. Lipopolysaccharide was used to induce inflammation in three kinds of corneal host cells in vitro, including corneal epithelial, stromal, and endothelial cells. Fungus was used to establish an animal model of corneal neovascularization in vivo. Tube formation and spheroid sprouting assays were used to evaluate the contribution of three kinds of corneal host cells to the degree of neovascularization under various stimuli. Matrix metalloproteinase (MMP)-2, alpha-crystallin A chain (CRYAA), galectin-8, Bcl-2, neuropilin-2, MMP-9 plasmids, and recombinant human fibronectin were used to identify the key proteins of corneal host cells involved in corneal inflammatory neovascularization. Findings: All three kinds of corneal host cells influenced corneal neovascularization to varying degrees. MMP-9 in human corneal epithelial cells, MMP-2, and CRYAA in human corneal stromal cells, and MMP-2 and galectin-8 in human corneal endothelial cells are potential key proteins that participate in corneal inflammatory neovascularization. Interpretation: Our data indicated that both the effects of key proteins and corneal host cells involved should be considered for the treatment of corneal inflammatory neovascularization. 

Retracted: No

PubPeer comments:
 In Figure 1A lines are included in some Western immuno blots (red arrows). I guess the lines mean a discontinuity and that two parts have been put together (not explained in the text). The problem is that the other blots and the actin loading blot do not have the same discontinuity, meaning that the samples are not run on the same gel, making the comparison between the immunoblots invalid. In Figure 6B you see the same problem. One of the bands are duplicated, but the loading control (and the rest of the blots) are different. You cannot just put pieces together as you like to make a story. Science supposed not to be like that. ![file] , 
 Dear Indigofera Tanganyikensis, Thank you for your comments on our recently published paper in EBioMedicine titled  Three kinds of corneal host cells contribute differently to corneal neovascularization EBioMedicine, Volume 44, 542 u2013 553. I am Haiyang Yu, the first author of this paper. The limited amounts of human corneal extract imposed technical challenges regarding the number of Western blots that could be performed. We were keen to maximize what could be experimentally obtained from the material we had, so some of the published blots were composites from multiple gels. Please refer to the attached images for a detailed explanation addressing the two comments you raised. Kind regards, Haiyang Yu![file] ![file] ![file] ![file] ![file] ![file] ![file] ,