Corneal Stromal Cell Growth on GelatinChondroitin Sulfate Scaffolds Modified at Different NHSEDC Molar Ratios.docVIP

Int. J. Mol. Sci. 2013, 14, 2036-2055; doi:10.3390/ijmOPEN ACCESS International Journal of Molecular Sciences ISSN 1422-0067 /journal/ijms Article Corneal Stromal Cell Growth on Gelatin/Chondroitin Sulfate Scaffolds Modified at Different NHS/EDC Molar Ratios Jui-Yang Lai 1,2,3 1 Institute of Biochemical and Biomedical Engineering, Chang Gung University, Taoyuan 33302, Taiwan; E-Mail: jylai@.tw; Tel.: +886-3-211-8800 (ext. 3598); Fax: +886-3-211-8668 2 Biomedical Engineering Research Center, Chang Gung University, Taoyuan 33302, Taiwan 3 Molecular Medicine Research Center, Chang Gung University, Taoyuan 33302, Taiwan Received: 5 November 2012; in revised form: 13 December 2012 / Accepted: 5 January 2013 / Published: 21 January 2013 Abstract: A nanoscale modification strategy that can incorporate chondroitin sulfate (CS) into the cross-linked porous gelatin materials has previously been proposed to give superior performance for designed corneal keratocyte scaffolds. The purpose of this work was to further investigate the influence of carbodiimide chemistry on the characteristics and biofunctionalities of gelatin/CS scaffolds treated with varying N-hydroxysuccinimide (NHS)/1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide hydrochloride (EDC) molar ratios (0-1) at a constant EDC concentration of 10 mM. Results of Fourier transform infrared spectroscopy and dimethylmethylene blue assays consistently indicated that when the NHS to EDC molar ratio exceeds a critical level (i.e., 0.5), the efficiency of carbodiimide-mediated biomaterial modification is significantly reduced. With the optimum NHS/EDC molar ratio of 0.5, chemical treatment could achieve relatively high CS content in the gelatin scaffolds, thereby enhancing the water content, glucose permeation, and fibronectin adsorption. Live/Dead assays and interleukin-6 mRNA expression a