Comparative Assessment of NHS/EDC and Sulfo-NHS/EDC Crosslinking Strategies for Cell-on-Chip Applications: Kinetics, Antifouling Properties, and Cytocompatibility - Dataset

Description

Abstract

The rapid advancement of microfluidic and cell-on-chip technologies is transforming biomedical research by enabling precise control over the cellular and measurement microenvironment. A critical requirement in these systems is the development of robust surface functionalization strategies that maintain long-term stability, antifouling properties, and cytocompatibility. This study systematically evaluates two carbodiimide-based conjugation systems for RGD coupling onto antifouling carboxybetaine-containing terpolymer brush surfaces, i.e., NHS/EDC and sulfo-NHS/EDC, by analyzing their activation kinetics, antifouling performance, and cellular compatibility with osteosarcoma (SaOS-2) cells. Fourier-transform infrared (FTIR) spectroscopy revealed distinct activation kinetics, with sulfo-NHS/EDC achieving 69% carboxylate conversion (1608 cm⁻¹ band reduction) within 30 min, compared to only 31% for NHS/EDC under identical conditions. Importantly, SaOS-2 cells exhibited enhanced spreading (37.1% vs. 16.7%) after 48 h of incubation on terpolymer brushes functionalized with RGD peptides via sulfo-NHS/EDC vs. via NHS/EDC. Both systems demonstrated exceptional antifouling properties up to 48 h. These findings establish sulfo-NHS/EDC as a superior strategy for long-term cell-on-chip applications, tissue-engineering scaffolds, and biosensor coatings that require stable biomolecular immobilization and sustained cytocompatibility.