Synthesis of Hyaluronic Acid Microsphere Crosslinked with Polyethylene Glycol Diglycidyl Ether Prepared by A Simple Fluidic Device

Hyaluronic acid (HA) microspheres (MSs) crosslinked with polyethylene glycol diglycidyl ether (PEGDE) are prepared using a simple fluidic device (SFD) to investigate the optimized parameters. A solution mixture of PEGDE in 2-methyl-1-propanol was prepared as a continuous phase in SFD. HA solutions of 1 wt% concentration were introduced into SFD as a discontinuous phase. The HA solution prepared by stirring for more than 48 h exhibited spherical MSs at the needle tip inside the ring cap. As the flow rate of the continuous phase increased from 0.7 to 1.9 mL/min, the diameter of the MS decreased from 173±36 ㎛ to 129±13 ㎛. Although the PEGDE concentration in the range of 0.2 to 1.8 vol% did not affect the diameter of the MS, the microstructure of MS, consisting of inner hollow void and wall, was changed. The inner void and wall size decreased and increased from 79.5 ㎛ to 57.2 ㎛ and from 10.3 ㎛ to 21.4 ㎛, respectively, with increasing PEGDE concentration from 0.2 vol% to 1.8 vol%. FT-IR peaks located around 2867 cm^-1 and 1088 cm^-1 indicated that the HA MS prepared at different PEGDE concentrations were chemically crosslinked. The HA MSs containing different PEGDE concentrations exhibited quantitative cell viability of more than 98%. L-929 cells adhered well to the HA MSs and proliferated continuously with increasing culture time to 48 h regardless of PEGDE concentration, implying that the HA MSs are clinically safe and effective.