Manufacturing of gellan gum microspheres to encapsulate the nanocarriers of doxorubicin for hepatoma therapy
Ming Wei Lee, Department of Medical Laboratory and Biotechnology, Chung Shan Medical University, Taichung, TaiwanMing Fang Hsu, Department of Medical Laboratory and Biotechnology, Chung Shan Medical University, Taichung, Taiwan
Hepatocellular carcinoma (HCC) is the most common primary malignancy of the liver and the third cause of cancer death. Transcatheter arterial chemoembolization has most widely been applied to hepatocellular carcinoma (HCC) for patients who are not eligible for surgery.
The study aims to design noval chemoembolization agent for hepatoma therapy. The chemoembolization agent is based on gellan gum microsphere which contain hyaluronan/polyethylenimine-doxorubicin nanoparticle (HH/PH-DOX). Hyaluronan conjugated with histidine (HH) and polyethylenimine conjugated with histidine (PH) were synthesized, 1H NMR spectrum showed that the peaks of HH were at 8.064 ppm (-N=CH-) and 6.979 ppm (-N-CH=C-) which were ascribed to the methenyl group located in imidazole group of histidine. The newly appeared peaks at 7.714 ppm (-N=CH-) and 6.959 ppm (-N-CH=C-) revealed that PH was successfully obtained. The delta-potential of HH and PH were -42.7 and 43.4 mV, respectively. The delta-potential of doxorubicin solution was -10 mV, when PH reacted with doxorubicin can form PH-DOX by covalent bonding formation and the delta-potential of PH-DOX was 33.6 mV. And then, PH-DOX mixed with HH at the w/w ratio of 1:4 to form HH/PH-DOX polyion complexes (delta-potential was -25.5mv). From the zeta potential analysis, we found that doxorubicin can encapsulate into HH/PH matrix successfully. The gellan gum microsphere contain HH/PH-DOX were fabricated using a W/O emulsion method, and the diameter of these microspheres was 300± 100 um.
The cellular uptake test, results showed that HH/PH-DOX (arrow) could be into HepG2 cells by endocytosis after co-incubation for 24h. From these datas, the gellan gum microsphere with HH/PH-DOX nano particles have the potential as the chemoembolization agent in the future.