Disulfide-crosslinked heparin-pluronic nanogels as a redox-sensitive nanocarrier for intracellular protein delivery

Abstract

Improving the efficacy of drug delivery via nanocarriers has been a major issue in the field of intravenous delivery. In this study, a polymeric nanogel was developed to enhance the stability, redox responsiveness, and the efficacy for intracellular protein delivery. The thiolated heparin-Pluronic conjugate was self-assembled and oxidized to form a disulfide-crosslinked nanogel network under a diluted aqueous condition. The disulfide-crosslinked heparin-Pluronic (DHP) nanogels with encapsulated RNase A were characterized by in vitro release and cytotoxicity tests depending on the existence of glutathione (GSH). The DHP nanogels exhibited reduced hydrodynamic size, higher encapsulation degree, and augmentable release responding to the GSH concentration. The ctotoxicity data confirmed that DHP nanogels were more effective for the intracellular delivery of RNase A compared to non-crosslinked nanogel.

Keywords

heparin nanogel glutathione RNase A heparin-Pluronic nanogels redox-sensitive nanocarrier intracellular protein delivery

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References

Verma IM and Sonia N. Gene therapy: promises, problems, and prospects. Nature 1997; 389: 239-242.

Leader B. , Baca QJ and Golan DE Protein therapeutics: a summary and pharmacological classification. Nat Rev Drug Disc 2008; 7: 21-39.

Tan CY and Kim YH Antiapoptotic Fusion Protein Delivery Systems. Macromol Res 2008; 16: 481-488.

Ohtsuka N. , Konno T. , Miyauchi Y. and Maeda H. Anticancer effects of arterial administration of the anticancer agent SMANCS with lipiodol on metastatic lymph nodes. Cancer 1987; 59: 1560-1565.

Duncan R.

Polymer conjugates for tumour targeting and intracytoplasmic delivery. The EPR effect as a common gateway?

PSTT 1999; 2: 441-449.

Chen DQ , Jiang XQ , Huang YY , Zhang C. and Ping Q. pH-sensitive mPEG-Hz-cholesterol conjugates as a liposome delivery system . J Bioact Compat Pol 2010; 25: 527-542.

Choi JH , Jang JY , Joung YK , Kwon MH and Park KD. Intracellular delivery and anti-cancer effect of self-assembled heparin-Pluronic nanogels with RNase A. J Control Release 2010; 147: 420-427.

Cho YW , Choi MY , Lee KH , et al. Cyclotriphosphazene-Pt-DACH conjugates with dipeptide Spacers for drug delivery systems. J Bioact Compat Pol 2010; 25: 274-291.

Park KM , Bae JW , Joung YK , Shin JW and Park KD Nanoaggregate of thermosensitive chitosan-pluronic for sustained release of hydrophobic drug. Colloid Surf B 2008; 63: 1-6.

10.

Batrakova EV and Kabanov AV Pluronic block copolymers: evolution of drug delivery concept from inert nanocarriers to biological response modifiers . J Control Release 2008; 130: 98-106.

11.

Capila I. and Linhardt RJ Heparin-protein interactions. Angew Chem Int Ed Engl 2002; 41: 391-412.

12.

Jee KS , Park HD , Kim YH , Shin JW and Park KD Heparin conjugated polylactide as a blood compatible material. Biomacromolecules 2004; 5: 1877-1881.

13.

Moon HT , Jeon OC , Byun Y. , Kim YJ and Lee YK Evaluation of the oral absorption of heparin conjugated with sodium deoxycholate as a facilitating agent in GI tract. Macromol Res 2009; 2: 79-83.

14.

Li HL , Ye KH , Zhang HW , Luo YR , Ren XD , Xiong AH , et al. Effect of heparin on apoptosis in human nasopharyngeal carcinoma CNE2 cells. Cell Res 2001; 11: 311-315.

15.

Bae KH , Mok H. and Park TG Synthesis, characterization, and intracellular delivery of reducible heparin nanogels for apoptotic cell death. Biomaterials 2008; 29: 3376-3383.

16.

Peptu CA , Buhus G. , Popa M. , Perichaud A. and Costin D. Double cross-linked chitosan-gelatin particulate systems for ophthalmic applications . J Bioact Compat Pol 2010; 25: 98-116.

17.

Etrych T. , Kovar L. , Subr V. , Braunova A. , Pechar M. , Chytil P. , et al. High-molecular-weight polymers containing biodegradable disulfide bonds: synthesis and in vitro verification of intracellular degradation. J Bioact Compat Pol 2010; 25: 5-26.

18.

Schmitz T. , Bravo-Osuna I. , Vauthier C. , Ponchel G. , Loretz B. and Bernkop-Schnurch A. Development and in vitro evaluation of a thiomerbased nanoparticulate gene delivery system. Biomaterials 2007; 28: 524-531.

19.

Ranucci E. , Suardi MA , Annunziata R. , Ferruti P. , Chiellini F. and Bartoli C. Poly(amidoamine) conjugates with disulfide-linked cholesterol pendants self-assembling into redox-sensitive nanoparticles. Biomacromolecules 2008; 9: 2693-2704.

20.

Neu M. , Germershaus O. , Mao S. , Voigt KH , Behe M. and Kissel T. Crosslinked nanocarriers based upon poly(ethylene imine) for systemic plasmid delivery: in vitro characterization and in vivo studies in mice. J Control Rel 2007; 118: 370-380.

21.

Loretz B. , Thaler M. and Schnurch AB Role of sulfhydryl groups in transfection? a case study with chitosan-nac nanoparticles. Bioconjugate Chem 2007; 18: 1028-1035.

22.

Smith PK , Mallia AK and Hermanson GT Toluidine blue. Anal Biochem 1980; 109: 466-473.

23.

Ellman GL Tissue sulfhydryl groups. Arch Biochem Biophys 1959; 82: 70-77.

24.

Allen C. , Maysinger D. and Eisenberg A. Nanoengineering block copolymer aggregates for drug delivery. Colloid Surf B: Biointerfaces 1999; 16: 3-27.

25.

Matousek J. , Pouckova P. , Hlouskova D. , Zadinova M. , Soucek J. and Skvor J. Effect of hyaluronidase and PEG chain conjugation on the biologic and antitumor activity of RNase A. J Control Rel 2004; 94: 401-410.

26.

Ahmed H. Estimation of protein, principles and reactions of protein extraction, purification, and characterization. Taylor & Francis, Inc, 2005, pp.49-60.

27.

Lee HY , Yu SA , Jeong KH and Kim YJ Poly(vinyl pyrrolidone) conjugated lipid system for the hydrophobic drug delivery . Macromol Res 2007; 15: 547-552.

28.

Edwards J. and Pearson R. The Factors determining nucleophilic reactivities. J Am Chem Soc 1962; 84: 16-24.

29.

Santos JR , Alves NM and Mano JF New thermo-responsive hydrogels based on poly (N-isopropylacrylamide)/hyaluronic acid- interpenetrated polymer networks: swelling properties and drug release studies. J Bioact Compat Pol 2010; 25: 169-184.

30.

Choi SH , Lee JH , Choi SM and Park TG Thermally reversible pluronic/ heparin nanocapsules exhibiting 1000-fold volume transition. Langmuir 2006; 22: 1758-1762.

31.

Prabha S. , Zhou WZ , Panyam J. and Labhasetwar V. Size-dependency of nanoparticlemediated gene transfection: studies with fractionated nanoparticles. Int J Pharm 2002; 244: 105-115.

32.

Jain RK Delivery of molecular and cellular medicine to solid tumors. J Control Release 1998; 53: 49-67.

33.

Griffiths G. , Back R. and Marsh M. A quantitative analysis of the endocytic pathway in baby hamster kidney cells . J Cell Biol 1989; 109: 2703-2720.

34.

Cavallaro G. , Campisi M. , Licciardi M. , Ogris M. and Giammona G. Reversibly stable thiopolyplexes for intracellular delivery of genes. J Control Release 2006; 115: 322-334.

35.

Kommareddy S. and Amiji M. Preparation and evaluation of thiol-modified gelatin nanoparticles for intracellular DNA delivery in response to glutathione . Bioconjugate Chem 2005; 16: 1423-1432.

36.

Kommareddy S. and Amiji M. Poly(ethylene glycol)-modified thiolated gelatin nanoparticles for glutathione-responsive intracellular DNA delivery. Nanomedicine 2007; 3: 32-42.

37.

Carlisle RC , Etrych T. , Briggs SS , Preece JA , Ulbrich K. and Seymour LW Polymer-coated polyethylenimine/DNA complexes designed for triggered activation by intracellular reduction. J Gene Med 2004; 6: 337-344.

38.

West KR and Otto S. Reversible covalent chemistry in drug delivery. Curr Drug Discovery Technol 2005; 2: 123-160.

39.

Kirpotin D. , Hong KL , Mullah N. , Papahadjopoulos D. and Zalipsky S. Liposomes with detachable polymer coating: destabilization and fusion of dioleoylphosphatidylethanolamine vesicles triggered by cleavage of surface-grafted poly(ethylene glycol). FEBS Lett 1996; 388: 115-118.

40.

Rosenberg HF RNase a ribonucleases and host defense: an evolving story. J Leukoc Biol 2008; 83: 1079-1087.