[Preparation of hispidulin chitosan microsphere and its antiproliferation effect in vitro]

Abstract

Hispidulin(HPDL) chitosan microspheres were prepared in this study to deliver HPDL to the lesion sitevia intravenous injection, and further evaluate their anticancer effects in vitro and the growth inhibition effect on A549 cells spheroids. HPDL chitosan microspheres were prepared by emulsion crosslinking method with chitosan as a drug carrier and the amount of HPDL was determined by high performance liquid chromatography (HPLC). The morphology of microspheres was observed under laser scanning confocal microscope. Additionally, the drug release amount of targeting microspheres was detected by dialysis method. Furthermore, the anti-proliferative effects against A549 lung cancer cells were tested by sulforhodamine B (SRB) method, and the effects of HPDL chitosan micrpsphereson early apoptosis of A549 cellswere determined by flow cytometry. A549 cells tumor spheroids were developed in vitro and then HPDL chitosan microspheres were added. On the 0, 1, 3, 7 d after adding the drugs, the inverted microscope was used to observe the mythologicaland volume changes of A549 cells spheroids. The encapsulation efficiency of HPDL chitosan microspheres was （75.32±0.52）%, and the drug loading amount was （7.76±0.67）%. Meanwhile, the microspheres were round shaped andhad smooth surface. The HPDL chitosan microspheres exhibited stronger inhibitory effects on A549 lung cancer cells. The results of flow cytometry indicated that, the early apoptosis rate of lung cancer A549 cells was （37.0±0.75）% at 24 h cells culture after drug administration. The volume of tumor spheroid was significantly inhibited, which had been shrunk by (50.09±11.06)% after the treatment by drug-loaded microsphere at day 7 as compared with blank group; meanwhile, the cells surface were obviously lysed. The preparation method in this research was simple and practicable, and the microspheres prepared with this method were round and smooth, with high encapsulation efficiency, which can significantly inhibit proliferation of lung adenocarcinoma A549 cells and induce cell apoptosis, and at the same time can cause lysisand death of A549 cell tumor spheroid.

Keywords: anticancer effects; apoptosis; hispidulin; microsphere; tumor spheroid.