Surface properties and bioactivity of PNIPAM-grafted-chitosan/chondroitin multilayers

Abstract

The thermoresponsive poly(N-isopropylacrylamide) (PNIPAM) is widely applied in the biomedical field particularly as thermoresponsive substrate for culture of cells. To be used as a stimuli-responsive coating for cell culture, combining PNIPAM with glycosaminoglycans might be an effective approach to improve its bioactivity. In this study, chitosan is grafted with PNIPAM moieties (PCHI) possessing a cloud point at 31 ​°C and used as a polycation to fabricate thermoresponsive polyelectrolyte multilayers (PEM) with the bioactive polyanion chondroitin sulfate (CS) at pH 4 by layer-by-layer technique. The in-situ investigation by surface plasmon resonance and quartz crystal microbalance with dissipation monitoring confirms that the formation of PEMs with CS can be achieved despite the bulky structure of PCHI at 25 ​°C. The stability of the PEMs is further improved at physiological pH 7.4 by chemical crosslinking using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide/N-hydroxysuccinimide. Moreover, these PEMs exhibit de-swelling and swelling ability with different surface wettability in response to temperature, which triggers the adsorption and desorption of adhesive protein vitronectin on the PEMs. At 37 ​°C, the PEMs containing PNIPAM particularly associated with CS terminal layer supports protein adsorption and consequently enhances cell adhesion using multipotent murine stem cells. Overall, due to improved stability, crosslinked PNIPAM-modified biogenic multilayers are cytocompatible and hold great potential as culture substrate for different tissue cells and application in tissue engineering.

Keywords

• Poly(N-isopropylacrylamide)
• Chitosan
• Chondroitin sulfate
• Polyelectrolyte multilayers
• Vitronectin adsorption
• Stem cell adhesion