Past Issue

Volume 18 , Number 2,Jul-Sep (Summer) 2016, Serial Number: 70, Pages: 135-148

In Vitro Co-Delivery Evaluation of Novel Pegylated Nano-Liposomal Herbal Drugs of Silibinin and Glycyrrhizic Acid (Nano-Phytosome) to Hepatocellular Carcinoma Cells

Mohammad Mahdi Ochi, Ph.D, 1, 2, Ghasem Amoabediny, Ph.D, 2, 3, *, Seyed Mahdi Rezayat, Ph.D, 4, Azim Akbarzadeh, Ph.D, 2, 5, Bahman Ebrahimi, Ph.D, 1,
Department of Life Science Engineering, Faculty of New Sciences and Technologies, University of Tehran, Tehran, Iran
Department of Nano Biotechnology, Research Center for New Technologies in Life Science Engineering, University of Tehran, Tehran, Iran
Department of Biotechnology and Pharmaceutical Engineering, School of Engineering, University of Tehran, Tehran, Iran
School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran
Pasteur Institute of Iran (IPI), Tehran, Iran
*Corresponding Address: P.O.Box: 14155-6619 Department of Biotechnology and Pharmaceutical Engineering School of Engineering University of Tehran Tehran Iran



This study aimed to evaluate a co-encapsulated pegylated nano-liposome system based on two herbal anti-tumor drugs, silibinin and glycyrrhizic acid, for delivery to a hepatocellular carcinoma (HCC) cell line (HepG2).

Materials and Methods

In this experimental study, co-encapsulated nano-liposomes by the thin layer film hydration method with HEPES buffer and sonication at 60% amplitude. Liposomes that co-encapsulated silibinin and glycyrrhizic acid were prepared with a specified molar ratio of dipalmitoylphosphatidylcholine (DPPC), cholesterol (CHOL), and methoxy-polyethylene glycol 2000 (PEG2000)–derived distearoyl phosphatidylethanolamine (mPEG2000-DSPE). We used the MTT technique to assess cytotoxicity for various concentrations of co-encapsulated nano-liposomes, free silibinin (25% w/v) and glycyrrhizic acid (75% w/v) on HepG2 and fibroblast cell lines over a 48-hour period.


Formulation of pegylated nano-liposomes showed a narrow size distribution with an average diameter of 46.3 nm. The encapsulation efficiency (EE) for silibinin was 24.37%, whereas for glycyrrhizic acid it was 68.78%. Results of in vitro cytotoxicity showed significantly greater co-encapsulated nano-liposomes on the HepG2 cell line compared to the fibroblast cell line. The half maximal inhibitory concentration (IC50) for co-encapsulated pegylated nanoliposomal herbal drugs was 48.68 µg/ml and free silibinin with glycyrrhizic acid was 485.45 µg/ml on the HepG2 cell line.


This in vitro study showed that nano-liposome encapsulation of silibinin with glycyrrhizic acid increased the biological activity of free drugs, increased the stability of silibinin, and synergized the therapeutic effect of silibinin with glycyrrhizic acid. The IC50 of the co-encapsulated nano-liposomes was lower than the combination of free silibinin and glycyrrhizic acid on the HepG2 cell line.