title: Intracellular delivery of heparin complexed with chitosan-g-poly(ethylene glycol) for inducing apoptosis.
abstract: OBJECTIVE
  Chitosan-g-PEG/heparin polyelectrolyte complex micelles were prepared for inducing apoptotic death of cancer cells.
  
  
  METHODS
  The cytotoxicity of polyelectrolyte complex micelles was evaluated by examining the growth inhibition of mouse melanoma B16F10 cells. Cellular uptake and apoptosis-inducing effect were investigated by confocal laser scanning microscopy and flow cytometric analysis, respectively.
  
  
  RESULTS
  The prepared polyelectrolyte complex micelles had a spherical shape with an average diameter of 162.8 +/- 18.9 nm. They were highly stable and well dispersed even in the presence of serum due to the presence of hydrophilic PEG shell layer surrounding the micelles. Moreover, they exhibited significantly higher cytotoxic activity against B16F10 cells compared to heparin or chitosan-g-PEG at the same concentration. The polyelectrolyte complex micelles were internalized by cancer cells to a greater extent than free heparin alone, indicating that the dramatic cell death was attributed to the increased cellular uptake of heparin. The internalized heparin was shown to induce apoptotic death of the cancer cells via a caspase-dependent pathway.
  
  
  CONCLUSIONS
  Nanosized and stable chitosan-g-PEG/heparin polyelectrolyte complex micelles were produced by a self-assembly process. The polyelectrolyte complex micelles facilitated the intracellular delivery of heparin, triggered the caspase activation, and consequently promoted apoptotic death of cancer cells.
authors.affiliation: Department of Biological Sciences, Korea Advanced Institute of Science and Technology, Daejeon, 305-701, South Korea.
authors.forename: Ki Hyun
authors.initials: KH
authors.lastname: Bae
authors.forename: Chang Won
authors.initials: CW
authors.lastname: Moon
authors.forename: Yuhan
authors.initials: Y
authors.lastname: Lee
authors.forename: Tae Gwan
authors.initials: TG
authors.lastname: Park
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents; D017209:Apoptosis / Q000187:drug effects*; D002240:Carbohydrate Sequence; D045744:Cell Line, Tumor; D048271:Chitosan / Q000737:chemistry*; D005079:Excipients / Q000737:chemistry*; D005434:Flow Cytometry; D006493:Heparin / Q000008:administration & dosage / Q000737:chemistry* / Q000494:pharmacology*; D008027:Light; D008546:Melanoma, Experimental / Q000188:drug therapy / Q000473:pathology; D051379:Mice; D008823:Micelles; D018625:Microscopy, Atomic Force; D011092:Polyethylene Glycols / Q000737:chemistry*; D012542:Scattering, Radiation
chemical_list: D000970:Antineoplastic Agents; D005079:Excipients; D008823:Micelles; D011092:Polyethylene Glycols; D006493:Heparin; D048271:Chitosan
references.citation: J Control Release. 1998 Jun;54(1):39-48
references.pmid: 9741902
references.citation: Eur J Pharm Biopharm. 1999 Sep;48(2):101-11
references.pmid: 10469928
references.citation: J Control Release. 2005 Nov 2;108(1):121-31
references.pmid: 16139916
references.citation: Nature. 2000 Oct 12;407(6805):770-6
references.pmid: 11048727
references.citation: Mol Biol Cell. 1993 Sep;4(9):953-61
references.pmid: 8257797
references.citation: Cytometry. 1992;13(8):795-808
references.pmid: 1333943
references.citation: Biomacromolecules. 2007 Jan;8(1):146-52
references.pmid: 17206800
references.citation: Nat Rev Drug Discov. 2003 May;2(5):347-60
references.pmid: 12750738
references.citation: Science. 1994 Mar 18;263(5153):1600-3
references.pmid: 8128245
references.citation: Proc Natl Acad Sci U S A. 2001 Mar 13;98(6):3352-7
references.pmid: 11248082
references.citation: J Gene Med. 2006 Apr;8(4):477-87
references.pmid: 16389625
references.citation: Pharm Res. 2007 Aug;24(8):1405-14
references.pmid: 17393074
references.citation: Pharm Res. 2007 Apr;24(4):705-14
references.pmid: 17318418
references.citation: Clin Cancer Res. 2005 Nov 15;11(22):8172-9
references.pmid: 16299249
references.citation: Biomacromolecules. 2001 Winter;2(4):1178-83
references.pmid: 11777390
references.citation: Chem Biol. 2004 Apr;11(4):487-98
references.pmid: 15123243
references.citation: J Biomed Mater Res A. 2007 Dec 1;83(3):806-12
references.pmid: 17559117
references.citation: Eur J Pharm Biopharm. 2004 Jan;57(1):35-52
references.pmid: 14729079
references.citation: Pharm Res. 2002 Jun;19(6):810-7
references.pmid: 12134951
references.citation: Biomaterials. 2007 Jun;28(16):2667-76
references.pmid: 17335894
references.citation: Nat Rev Cancer. 2002 Jul;2(7):521-8
references.pmid: 12094238
references.citation: Biomaterials. 2008 Aug;29(23):3376-83
references.pmid: 18474396
references.citation: Bioconjug Chem. 2006 Sep-Oct;17(5):1209-18
references.pmid: 16984130
references.citation: Adv Drug Deliv Rev. 2001 Nov 5;52(2):117-26
references.pmid: 11718935
references.citation: Adv Drug Deliv Rev. 2006 Jul 7;58(4):467-86
references.pmid: 16781003
references.citation: J Control Release. 2004 Nov 5;100(1):5-28
references.pmid: 15491807
references.citation: Annu Rev Biochem. 1999;68:383-424
references.pmid: 10872455
references.citation: Bioconjug Chem. 1995 Jan-Feb;6(1):7-20
references.pmid: 7711106
references.citation: J Control Release. 2006 Apr 10;111(3):299-308
references.pmid: 16481062
references.citation: Science. 2003 Apr 25;300(5619):595-6
references.pmid: 12714733
references.citation: Adv Drug Deliv Rev. 2002 Feb 21;54(2):203-22
references.pmid: 11897146
references.citation: Nat Rev Drug Discov. 2003 Mar;2(3):214-21
references.pmid: 12612647