PCL microspheres tailored with carboxylated poly(glycidyl methacrylate)–REDV conjugates as conducive microcarriers for endothelial cell expansion

Abstract

Microcarrier cell culture systems provide one of the most promising techniques for cell amplification due to their high surface area-to-volume ratio. In this study, biodegradable polycaprolactone (PCL) microspheres tethered with carboxylated poly(glycidyl methacrylate)–REDV conjugates were developed by a combination of surface-initiated atom transfer radical polymerization (ATRP) and azide–alkyne click chemistry as conducive microcarriers for rapid cell expansion of human umbilical vein endothelial cells (HUVECs). Azido-terminated poly(glycidyl methacrylate) (PGMA-N₃) brushes were grafted onto the PCL microspheres by surface-initiated ATRP. Subsequent carboxylation of PGMA-N₃ brushes was accomplished by azide–alkyne click reaction with hexynoic acid. REDV peptides were covalently conjugated to the pendent carboxyl groups on the side chain of carboxylated PGMA-COOH brushes via carbodiimide chemistry to enhance the cytocompatibility of the three-dimension (3D) PCL scaffolding system. Success in each functionalization step was ascertained by the measurement of attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR), X-ray photoelectron spectroscopy (XPS), scanning electron microscopy (SEM) and wet laser particle size analysis. In vitro cell-loading assay of HUVECs demonstrated a significant improvement of cell adhesion and proliferation on the REDV-immobilized PCL microsphere surfaces, and a confluent layer of HUVECs was formed after 7 days of incubation. The highly biocompatible and transportable nature of functionalized PCL microcarriers offers significant potential as a cell expansion platform.