J Korean Neurosurg Soc.  2020 Nov;63(6):698-706. 10.3340/jkns.2019.0206.

Effect of Podophyllotoxin Conjugated Stearic Acid Grafted Chitosan Oligosaccharide Micelle on Human Glioma Cells

• Wang GH
• Shen HP
• Huang X
• Jiang XH
• Jin CS
• Chu ZM

• KMID: 2508599
• DOI: http://doi.org/10.3340/jkns.2019.0206

Abstract

Objective
: To study the physiochemical characteristics of podophyllotoxin (PPT) conjugated stearic acid grafted chitosan oligosaccharide micelle (PPT-CSO-SA), and evaluate the ability of the potential antineoplastic effects against glioma cells.
Methods
: PPT-CSO-SA was prepared by a dialysis method. The quality of PPT-CSO-SA including micellar size, zeta potential, drug encapsulation efficiency and drug release profiles was evaluated. Glioma cells were cultured and treated with PPT and PPT-CSO-SA. The ability of glioma cells to uptake PPT-CSO-SA was observed. The proliferation of glioma cells was determined by 3-[4, 5-dimethyl-2-thiazolyl]-2, 5-diphenyl-2H-tetrazolium bromide (MTT) assay. The apoptosis and morphology of U251 cells were observed by 4’,6-Diamidino-2-phenylindole dihydrochloride (DAPI) dye staining. Cell cycle analysis was performed by flow cytometry. The migration ability of U251 cells was determined by wound healing test.
Results
: PPT-CSO-SA had nano-level particle size and sustained release property. The encapsulation efficiency of drug reached a high level. The cellular uptake percentage of PPT in glioma cells was lower than that of PPT-CSO-SA (p<0.05). The inhibitory effect of PPT-CSO-SA on glioma cells proliferation was significantly stronger than that of PPT (p<0.05). The morphologic change of apoptosis cell such as shrinkage, karyorrhexis and karyopyknosis were observed. The percentage of U251 cells in G2/M phase increased significantly in the PPT-CSO-SA group compared with PPT group (p<0.05). Compared with the PPT group, the cell migration ability of the PPT-CSO-SA group was significantly inhibited after 12 and 24 hours (p<0.05).
Conclusion
: PPT-CSO-SA can effectively enhance the glioma cellular uptake of drugs, inhibit glioma cells proliferation and migration, induce G2/M phase arrest of them, and promote their apoptosis. It may be a promising anti-glioma nano-drug.

Keyword

Podophyllotoxin; Nanoparticles; Glioma; Antineoplastic agents

Figure

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  Fig. 1. Drug release profiles from 5%, 10%, 15%, and 20% PPT-CSO-SA in PBS (pH 7.4) solution. PPT : podophyllotoxin, CSO : chitosan oligosaccharide, SA : stearic acid.

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  Fig. 2. Cellular uptake percentages of PPT and PPT-CSO-SA in different glioma cells against incubation time : (A) U251, (B) U87. Cellular uptake percentage of PPT in PPT-CSO-SA by U251, U87 cells within different incubation time (C). PPT : podophyllotoxin, CSO : chitosan oligosaccharide, SA : stearic acid.

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  Fig. 3. Apoptosis of U251 cells induced by PPT-CSO-SA (A : DMSO was used as control; B and C : 20 ug/mL and 50 ug/mL PPT-CSO-SA; D : 50 ug/mL PPT). PPT : podophyllotoxin, CSO : chitosan oligosaccharide, SA : stearic acid.

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  Fig. 4. Compared with the PPT group, the cell migration ability of the PPT-CSO-SA group was significantly inhibited after 12 and 24 hours. PPT : podophyllotoxin, CSO : chitosan oligosaccharide, SA : stearic acid.

Reference

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