Dataset Information

Extracellular Matrix Protein Coating of Processed Fish Scales Improves Human Corneal Endothelial Cell Adhesion and Proliferation.

ABSTRACT: Purpose:Corneal transplantation can treat corneal endothelial diseases. Implanting cultivated human corneal endothelial cells (HCECs) via a cell carrier has clinical value as an alternative therapeutic strategy. This study was performed to compare the feasibility of fish scales and other biomaterials (gelatin and chitosan) as cell carriers and to investigate the effects of an extracellular matrix (ECM) protein coating to improve the cytocompatibility of fish scales. Methods:The physical properties of gelatin, chitosan, and fish scales were compared. Immortalized HCECs (B4G12) were cultured on processed fish scales, which were coated with fibronectin, laminin, collagen IV, or FNC Coating Mix. Cell attachment and proliferation were evaluated by immunofluorescence, cell counting, and bromodeoxyuridine (BrdU) labeling assays. Immunoblots were used to examine the expression levels of integrin-linked kinase (ILK), phosphate-ILK, ?-catenin, p63, and cell cycle mediators (cyclin D1 and p27Kip1). Results:The transparency of processed fish scales was better than that of chitosan, while the strength was higher than that of gelatin. The laminin, collagen IV, and FNC coatings facilitated B4G12 cell adhesion and proliferation, while fibronectin only facilitated cell adhesion. The laminin, collagen IV, and FNC coatings also upregulated phosphate-ILK and p63 expression. In addition, the FNC coating activated cell cycle mediators. Conclusion:ECM protein-coated processed fish scales can serve as a novel cell carrier to facilitate the development of HCEC transplantation. Translational Relevance:Improving the physical properties and cytocompatibility of fish scales as a cell carrier will facilitate the transplantation of HCECs into corneas for the purpose of cell therapy.

SUBMITTER: Hsueh YJ

PROVIDER: S-EPMC6543859 | biostudies-literature | 2019 May

REPOSITORIES: biostudies-literature

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Extracellular Matrix Protein Coating of Processed Fish Scales Improves Human Corneal Endothelial Cell Adhesion and Proliferation.

Hsueh Yi-Jen YJ Ma David Hui-Kang DH Ma Kathleen Sheng-Chuan KS Wang Tze-Kai TK Chou Cheng-Hung CH Lin Chien-Cheng CC Huang Min-Chang MC Luo Li-Jyuan LJ Lai Jui-Yang JY Chen Hung-Chi HC

Translational vision science & technology 20190530 3

<h4>Purpose</h4>Corneal transplantation can treat corneal endothelial diseases. Implanting cultivated human corneal endothelial cells (HCECs) via a cell carrier has clinical value as an alternative therapeutic strategy. This study was performed to compare the feasibility of fish scales and other biomaterials (gelatin and chitosan) as cell carriers and to investigate the effects of an extracellular matrix (ECM) protein coating to improve the cytocompatibility of fish scales.<h4>Methods</h4>The ph ...[more]

PMID: 31171994

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Project description:There is limited information on region-specific gene expression in the human corneal stroma. In this study, we aimed to investigate the expression profile of the extracellular matrix and adhesion molecules in the normal corneal stroma using laser capture microdissection (LCM) and molecular techniques.Frozen sections of human cornea without ocular disease were used to isolate the central and peripheral corneal stromal keratocytes by LCM. RNA was extracted from LCM-captured tissues and the RT2 Profiler PCR Arrays were used to examine the expression profile of extracellular matrix and adhesion molecules in the central and peripheral stroma. Real-time quantitative PCR was used to quantify gene expression. Proteomic and western blotting (WB) analyses were performed to confirm gene expression at protein level. Function association network was generated via the web tools String and Cytoscape.The gene expression profiling demonstrated that 35 out of the 84 extracellular matrix and adhesion molecules represented in the array were expressed in stromal keratocytes. Among them, 24 genes were not previously described in the corneal stroma. Two genes were found more abundantly expressed in the central stroma than in the periphery: TGFBI, COL6A2 (p < 0.05). ADAMTS13 was detected only in the central stroma. Proteomics and WB analysis confirmed the expression of 10 genes. Functional analysis revealed that most identified genes were presented in a core cluster that had multiple and strong associations with other genes.This study identified genes not previously described in the corneal stroma, revealed regional differences in gene expression between central and peripheral stroma, and also detected some interesting candidate genes that may play important roles in corneal function. These observations serve as the foundation to further investigate the molecular and cellular mechanisms regulating the pathogenesis of regional corneal stromal disorders such as keratoconus.

Dataset Information

Extracellular Matrix Protein Coating of Processed Fish Scales Improves Human Corneal Endothelial Cell Adhesion and Proliferation.

Publications

Extracellular Matrix Protein Coating of Processed Fish Scales Improves Human Corneal Endothelial Cell Adhesion and Proliferation.

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