Fateme Haghiralsadat1,2, Ghasem Amoabediny3, Samira Naderinezhad4, Kamran Nazmi5, Jantine Posthuma De Boer6, Behrouz Zandieh-Doulabi7, Tymour Forouzanfar2, Marco N Helder2. 1. Department of Life Science Engineering, Faculty of New Sciences & Technologies, University of Tehran, Tehran, Iran. 2. Department of Oral & Maxillofacial Surgery, VU University Medical Center, MOVE Research Institute Amsterdam, Amsterdam, The Netherlands. 3. Department of Biotechnology and Pharmaceutical Engineering, Faculty of Chemical Engineering, School of Engineering, University of Tehran, Tehran, Iran. amoabedini@ut.ac.ir. 4. Department of Biotechnology and Pharmaceutical Engineering, Faculty of Chemical Engineering, School of Engineering, University of Tehran, Tehran, Iran. 5. Department of Oral Biochemistry, Academic Centre for Dentistry Amsterdam (ACTA), University of Amsterdam and Vrije Univrsiteit Amsterdam, Amsterdam Movement Sciences, Amsterdam, The Netherlands. 6. Department of Orthopedic Surgery, VU University Medical Center, Amsterdam Movement Sciences, Amsterdam, Netherlands. 7. Oral Cell Biology and Functional Anatomy, Academic Centre for Dentistry Amsterdam (ACTA), University of Amsterdam and Vrije Universiteit Amsterdam, Amsterdam, The Netherlands.
Abstract
PURPOSE: To employ Doxorubicin-loaded liposomes, modified with YSA-peptide to target EphA2, to reduce adverse effects against primary bone cells and maximize toxicity against Saos-2 osteosarcoma cells. METHODS: PEGylated liposomes were prepared by thin film method using Dipalmitoylphosphatidylcholine (DPPC), cholesterol and distearylphosphatidylethanolamine-polyethyleneglycol conjugate (DSPE-mPEG) in 67.9:29.1:3 M ratios, and loaded with DOX (L-DOX) by pH-gradient method. Targeted liposomes (YSA-L-DOX), were prepared by conjugating YSA-peptide to DSPE-mPEG. Liposomes were physicochemically characterized and tested in cellular toxicity assays. RESULTS: YSA conjugation efficiency was >98%. Size and polydispersity index of both L-DOX and YSA-L-DOX were around 88 nm and 0.188, respectively. Both had similar zeta potential, and 85% DOX loading efficiencies. DOX release kinetics followed the Korsmeyer-Peppa model, and showed comparable release for both formulations from 1-8 h, and a plateau of 29% after 48 h. Both formulations could be stably stored for ≥6 months at 4°C in the dark. Toxicity assays showed a significant 1.91-fold higher cytotoxicity compared to free DOX in the Saos-2 cells, and 2-fold lesser toxicity in primary bone cells compared to the Saos-2 cells. Cellular uptake studies showed higher and more nuclear uptake in YSA-L-DOX compared to L-DOX treated cells. CONCLUSIONS: YSA-L-DOX vesicles might be effective for targeted treatment of osteosarcoma.
PURPOSE: To employ Doxorubicin-loaded liposomes, modified with YSA-peptide to target EphA2, to reduce adverse effects against primary bone cells and maximize toxicity against Saos-2 osteosarcoma cells. METHODS: PEGylated liposomes were prepared by thin film method using Dipalmitoylphosphatidylcholine (DPPC), cholesterol and distearylphosphatidylethanolamine-polyethyleneglycol conjugate (DSPE-mPEG) in 67.9:29.1:3 M ratios, and loaded with DOX (L-DOX) by pH-gradient method. Targeted liposomes (YSA-L-DOX), were prepared by conjugating YSA-peptide to DSPE-mPEG. Liposomes were physicochemically characterized and tested in cellular toxicity assays. RESULTS: YSA conjugation efficiency was >98%. Size and polydispersity index of both L-DOX and YSA-L-DOX were around 88 nm and 0.188, respectively. Both had similar zeta potential, and 85% DOX loading efficiencies. DOX release kinetics followed the Korsmeyer-Peppa model, and showed comparable release for both formulations from 1-8 h, and a plateau of 29% after 48 h. Both formulations could be stably stored for ≥6 months at 4°C in the dark. Toxicity assays showed a significant 1.91-fold higher cytotoxicity compared to free DOX in the Saos-2 cells, and 2-fold lesser toxicity in primary bone cells compared to the Saos-2 cells. Cellular uptake studies showed higher and more nuclear uptake in YSA-L-DOX compared to L-DOX treated cells. CONCLUSIONS:YSA-L-DOX vesicles might be effective for targeted treatment of osteosarcoma.
Authors: Gennady N Machak; Sergey I Tkachev; Yuriy N Solovyev; Pavel A Sinyukov; Stanislav M Ivanov; Natalya V Kochergina; Alexey D Ryjkov; Valery V Tepliakov; Benjamin Y Bokhian; Valeria V Glebovskaya Journal: Mayo Clin Proc Date: 2003-02 Impact factor: 7.616
Authors: Gergely Szakács; Jill K Paterson; Joseph A Ludwig; Catherine Booth-Genthe; Michael M Gottesman Journal: Nat Rev Drug Discov Date: 2006-03 Impact factor: 84.694