Sequential robust design methodology and X-ray photoelectron spectroscopy to analyze the grafting of hyaluronic acid to glass substrates.
Sequential Robust Design experiments and X-ray photoelectron spectroscopic (XPS) studies were performed to examine the immobilization of hyaluronic acid (HA) on glass substrates chemisorbed with N-(2-aminoethyl)-3-aminopropyl-trimethoxysilane (EDS). Numerous reaction conditions were investigated, including the concentrations of 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC), N-hydroxysulfosuccinimide (Sulfo-NHS), and HA, and the reaction buffer type, concentration, and pH. The elemental surface compositions of carbon and silicon (C/Si ratio) were used to assess the extent of HA immobilization, leading to the identification of critical HA-binding reaction conditions and the determination of an optimum surface chemistry. The optimum chemistry consisted of 200 mM EDC, 50 mM Sulfo-NHS, 10 mM N-(2-hydroxyethyl)piperazine-N'-(2-ethanesulfonic acid) (HEPES) buffer at a pH of 7.0, and 3 mg/mL HA. This work emphasizes the advantages of using Robust Design methods over traditional statistical experimental design, particularly when large numbers of variables are examined and costly analytical techniques are employed.
Department of Biomedical Engineering, Robert R. McCormick School of Engineering and Applied Sciences, Northwestern University, Evanston, Illinois 60208, USA.
Ranee A
RA
Stile
Thomas A
TA
Barber
David G
DG
Castner
Kevin E
KE
Healy
D000268:Adhesiveness; D001672:Biocompatible Materials / Q000737:chemistry*; D005898:Glass / Q000737:chemistry*; D006820:Hyaluronic Acid / Q000737:chemistry*; D008422:Materials Testing; D011474:Prosthesis Design / Q000379:methods; D012826:Silicone Elastomers; D013057:Spectrum Analysis; D013499:Surface Properties; D014965:X-Rays
D001672:Biocompatible Materials; D012826:Silicone Elastomers; C006194:aminoethyl-aminopropyl-trimethoxysilane; D006820:Hyaluronic Acid