Comparison of the Effects of Different Coating Matrices on Cell Binding and Nitric Oxide-Mediated Protein S-Nitrosylation in Endothelial Cells under Shear Flow

Title:Comparison of the Effects of Different Coating Matrices on Cell Binding and Nitric Oxide-Mediated Protein S-Nitrosylation in Endothelial Cells under Shear Flow

Volume: 21

Author(s): Ming-Chung Lin, Ming-Wei Lin, Erna Sulistyowati, Ching-Chieh Kao, Chung-Jung Liu, Shu-Ping Huang, Sodio C. N. Hsu and Bin Huang*

Affiliation:

• Regenerative Medicine and Cell Therapy Research Center, Kaohsiung Medical University, Kaohsiung 80708, Taiwan
• Department of Biomedical Science and Environmental Biology, College of Life Science, Kaohsiung Medical University, Kaohsiung 80708, Taiwan
• Division of Gastroenterology, Department of Internal Medicine, Kaohsiung Medical University Chung-Ho Memorial Hospital, Kaohsiung 80708, Taiwan
• Department of Medical Research, Kaohsiung Medical University Chung-Ho Memorial Hospital, Kaohsiung 80708, Taiwan
• Department of Medical Research, Kaohsiung Medical University Chung-Ho Memorial Hospital, Kaohsiung 80708, Taiwan

Abstract: Objective: Shear flow is a mechanical signal regulating the function of Endothelial Cells (ECs). The present study aimed to investigate the effects of different matrices on cell binding, Nitric Oxide (NO) production, protein S-nitrosylation, expression of adhesion proteins, ROS generation, and cell viability in ECs under shear flow.

Methods: The ECs growing on glass slides separately coated with poly-L-lysine (p-Lys), collagen (Colla), fibronectin (Fibro), and a combined matrix (Colla+Fibro) were exposed to shear flow (25 dyne/cm2) for 0, 1, 4, 8 h. The number of ECs remaining attached on the glass slide was calculated. The expressions of endothelial Nitric Oxide Synthase (eNOS), peNOSS1177, VE-cadherin, FAK, and S-nitrosylated proteins were investigated by western blotting. The production of Nitric Oxide (NO) was measured by a specific reagent. Finally, the levels of ROS and cell viability were monitored.

Results: Under a constant shear flow for 1 h, the physiological responses of ECs were similar between these four matrices. When shear flow was extended to 4 and 8 h, higher cell binding, elevated NO production, increased S-nitrosylated proteins, enhanced expressions of FAK and VE-cadherin, mildly accumulated ROS, and cell death were observed in the matrix of Fibro and Colla+Fibro.

Conclusion: We have concluded fibronectin to be the optimal matrix facilitating NO-mediated Snitrosylation that might be essential for superior binding efficiency, thereby preventing the stripping of ECs under shear flow. The results can be broadly applied to diverse biomechanical studies.

Cite this article as:

Lin Ming-Chung, Lin Ming-Wei, Sulistyowati Erna, Kao Ching-Chieh, Liu Chung-Jung, Huang Shu-Ping, Hsu C. N. Sodio and Huang Bin*, Comparison of the Effects of Different Coating Matrices on Cell Binding and Nitric Oxide-Mediated Protein S-Nitrosylation in Endothelial Cells under Shear Flow, Current Proteomics 2024; 21 () . https://dx.doi.org/10.2174/0115701646300960240606093535