Hyaluronic acid/Fluorouracil

Lysozyme (LYS) and hyaluronan with low (HA1: 3 kDa), medium (HA2: 120 kDa), and high (HA3: 1200 kDa) molecular weights were used to fabricate lysozyme-hyaluronan colloidal nanoparticles using a green self-assembly method. Fourier transform infrared spectroscopy indicated that hydrogen bonding, hydrophobic and electrostatic interactions promoted the formation of the colloidal nanoparticles. The hydrophobic area of prepared colloidal nanoparticles was quantified using a pyrene fluorescent probe, and the results showed that the LYS-HA3 nanoparticles had the strongest hydrophobic capacity. Furthermore, 5-fluorouracil (5-Fu) was used to evaluate encapsulation performance, demonstrating that the LYS-HA3 nanoparticles had the highest encapsulation ability (>90 %). All prepared 5-Fu-loaded lysozyme-hyaluronan (5-Fu@LYS-HA) colloidal nanoparticles exhibited excellent long-term storage stability at 4 °C for 60 days. Cellular uptake and in vitro release results indicated that the LYS-HA2 nanoparticles exhibited the highest cellular uptake efficiency, and the LYS-HA3 nanoparticles had the best slow-release effect, while the release process was mainly controlled by the combination of Fickian diffusion and structural relaxation, respectively. This study demonstrates the influence of molecular weight on the conformational and structural properties of colloidal nanoparticles, which has implications for the design of insoluble drug self-assembly systems.

The use of liposomes as drug carriers improves the therapeutic efficacy of anticancer drugs, while at the same time reducing side effects. Hyaluronic acid (HA) is recognized by the CD44 receptor, which is overexpressed in many cancer cells. In this study, we developed HA‐modified liposomes encapsulating 5‐fluorouracil (5‐FU) and tested them against a CD44 expressing colorectal cell line (HT29) and a non‐CD44 expressing hepatoma cell line. The average size of 5‐FU‐lipo and 5‐FU‐lipo‐HA nanoparticles were 112 ± 28 and 144 ± 77 nm, respectively. The 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium (MTT) assay showed selective cancer cell death depending on the CD44 expression in a time‐dependent manner. Apoptosis assays and cell‐cycle analysis indicated that G0/G1 arrest occurred. The colony formation study revealed that cells treated with 5‐FU‐lipo and 5‐FU‐lipo‐HA had reduced colony formation. Quantitative reverse‐transcription polymerase chain reaction study showed that the oncogenic messenger RNA and microRNA levels were significantly reduced in the 5‐FU‐lipo‐HA‐treated group, while tumor suppressors were increased in that group. We suggest that optimal targeted delivery and release of 5‐FU into colorectal cancer cells, renders them susceptible to apoptosis, cell‐cycle arrest, and decreased colony formation.