AAPS PharmSciTech

, Volume 19, Issue 7, pp 3210–3218 | Cite as

Evaluating the Accelerated Blood Clearance Phenomenon of PEGylated Nanoemulsions in Rats by Intraperitoneal Administration

  • Yuqing Su
  • Mengyang Liu
  • Kaifan Liang
  • Xinrong Liu
  • Yanzhi SongEmail author
  • Yihui DengEmail author
Research Article


The accelerated blood clearance (ABC) phenomenon is induced by repeated intravenous injection of stealth polyethylene glycol (PEG) nanocarriers and appears as the alteration of the pharmacokinetics and biodistribution of the second administration. Nevertheless, there is no any report about the ABC phenomenon induced by intraperitoneal administration of PEGylated nanocarriers. In this study, we firstly observed whether the ABC phenomenon is induced with PEGylated nanoemulsion at the dose of 0.5~100 μmol phospholipid·kg−1 by intraperitoneal/intravenous injections in rats. The PEG (molecule weight, 2000)-modified nanoemulsions PE-B and PE in which fluorescence indicator dialkylcarbocyanines (DiR) is encapsulated by PE-B were prepared for this work. The pharmacokinetics of the first injected PE via veins or peritoneal cavity features different variation trends. Moreover, the tissue distributions (in vivo or in vitro) of the first injected PE by intraperitoneal injection reveals that the PE gains access to the whole lymphatic circulatory system. Furthermore, our results demonstrate that the ABC phenomenon can be induced by intraperitoneal administration PE-B and present obvious changes with varying PE-B concentration 0.5~100 μmol phospholipid·kg−1. Moreover, an interesting point is that the ABC phenomenon induced by intraperitoneal injected PE-B can be significantly inhibited by intraperitoneal pre-injection of distilled water. For understanding this issue clear, we studied the production of anti-PEG IgM and the characteristic morphologies of immune cells. We observed that the mast cells in peritoneal cavity exhibit rapid depletion in response to the intraperitoneal pre-injection of distilled water, while the anti-PEG IgM secretes at the same level.


PEGylated emulsions ABC phenomenon intraperitoneal injection anti-PEG IgM mast cells 


Funding Information

This research was aided financially by the National Natural Science Foundation of China (Grant Nos. 81072602, 81373334).

Supplementary material

12249_2018_1120_MOESM1_ESM.docx (797 kb)
ESM 1 (DOCX 796 kb)


  1. 1.
    Soppimath KS, Aminabhavi TM, Kulkarni AR, Rudzinski WE. Biodegradable polymeric nanoparticles as drug delivery devices. J Control Release. 2001;70(1–2):1–20.CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Singh R, Lillard JW Jr. Nanoparticle-based targeted drug delivery. Exp Mol Pathol. 2009;86(3):215–23.CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Mainardes RM, Silva LP. Drug delivery systems: past, present, and future. Curr Drug Targets. 2004;5(5):449–55.CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Bunker A, Magarkar A, Viitala T. Rational design of liposomal drug delivery systems, a review: combined experimental and computational studies of lipid membranes, liposomes and their PEGylation. Biochim Biophys Acta. 2016;1858(10):2334–52.CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Jokerst JV, Lobovkina T, Zare RN, Gambhir SS. Nanoparticle PEGylation for imaging and therapy. Nanomedicine. 2011;6(4):715–28.CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Dams ET, Laverman P, Oyen WJ, Storm G, Scherphof GL, van der Meer JW, et al. Accelerated blood clearance and altered biodistribution of repeated injections of sterically stabilized liposomes. J Pharmacol Exp Ther. 2000;292(3):1071–9.PubMedGoogle Scholar
  7. 7.
    Laverman P, Carstens MG, Boerman OC, Dams ETM, Oyen WJ, van Rooijen N, et al. Factors affecting the accelerated blood clearance of polyethylene glycol-liposomes upon repeated injection. J Pharmacol Exp Ther. 2001;298(2):607–12.PubMedGoogle Scholar
  8. 8.
    Ishida T, Maeda R, Ichihara M, Mukai Y, Motoki Y, Manabe Y, et al. The accelerated clearance on repeated injection of pegylated liposomes in rats: laboratory and histopathological study. Cell Mol Biol Lett. 2002;7(2):286.PubMedGoogle Scholar
  9. 9.
    Lila ASA, Kiwada H, Ishida T. The accelerated blood clearance (ABC) phenomenon: clinical challenge and approaches to manage. J Control Release. 2013;172(1):38–47.CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Im H-J, England CG, Feng L, Graves SA, Hernandez R, Nickles RJ, et al. Accelerated blood clearance phenomenon reduces the passive targeting of PEGylated nanoparticles in peripheral arterial disease. ACS Appl Mater Interfaces. 2016;8(28):17955–63.CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Ishihara T, Takeda M, Sakamoto H, Kimoto A, Kobayashi C, Takasaki N, et al. Accelerated blood clearance phenomenon upon repeated injection of PEG-modified PLA-nanoparticles. Pharm Res. 2009;26(10):2270–9.CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Zhao Y, Wang L, Yan M, Ma Y, Zang G, She Z, et al. Repeated injection of PEGylated solid lipid nanoparticles induces accelerated blood clearance in mice and beagles. Int J Nanomedicine. 2012;7:2891.PubMedPubMedCentralGoogle Scholar
  13. 13.
    Koide H, Asai T, Hatanaka K, Urakami T, Ishii T, Kenjo E, et al. Particle size-dependent triggering of accelerated blood clearance phenomenon. Int J Pharm. 2008;362(1–2):197–200.CrossRefGoogle Scholar
  14. 14.
    Shiraishi K, Yokoyama M. Polymeric micelles possessing polyethyleneglycol as outer shell and their unique behaviors in accelerated blood clearance phenomenon. Biol Pharm Bull. 2013;36(6):878–82.CrossRefGoogle Scholar
  15. 15.
    Środa K, Rydlewski J, Langner M, Kozubek A, Grzybek M, Sikorski AF. Repeated injections of PEG-PE liposomes generate anti-PEG antibodies. Cell Mol Biol Lett. 2005;10:37–47.PubMedGoogle Scholar
  16. 16.
    Ishida T, Ichihara M, Wang X, Kiwada H. Spleen plays an important role in the induction of accelerated blood clearance of PEGylated liposomes. J Control Release. 2006;115(3):243–50.CrossRefGoogle Scholar
  17. 17.
    Ishida T, Ichihara M, Wang X, Yamamoto K, Kimura J, Majima E, et al. Injection of PEGylated liposomes in rats elicits PEG-specific IgM, which is responsible for rapid elimination of a second dose of PEGylated liposomes. J Control Release. 2006;112(1):15–25.CrossRefGoogle Scholar
  18. 18.
    Shimizu T, Ishida T, Kiwada H. Transport of PEGylated liposomes from the splenic marginal zone to the follicle in the induction phase of the accelerated blood clearance phenomenon. Immunobiology. 2013;218(5):725–32.CrossRefGoogle Scholar
  19. 19.
    Ishida T, Wang X, Shimizu T, Nawata K, Kiwada H. PEGylated liposomes elicit an anti-PEG IgM response in a T cell-independent manner. J Control Release. 2007;122(3):349–55.CrossRefGoogle Scholar
  20. 20.
    Ishida T, Kashima S, Kiwada H. The contribution of phagocytic activity of liver macrophages to the accelerated blood clearance (ABC) phenomenon of PEGylated liposomes in rats. J Control Release. 2008;126(2):162–5.CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Shimizu T, Mima Y, Hashimoto Y, Ukawa M, Ando H, Kiwada H, et al. Anti-PEG IgM and complement system are required for the association of second doses of PEGylated liposomes with splenic marginal zone B cells. Immunobiology. 2015;220(10):1151–60.CrossRefPubMedPubMedCentralGoogle Scholar
  22. 22.
    Wang L, Su Y, Wang X, Liang K, Liu M, Tang W, et al. Effects of complement inhibition on the ABC phenomenon in rats. Asia J Pharmaceut Sci. 2017;12(3):250–8.Google Scholar
  23. 23.
    Zhao Y, Wang C, Wang L, Yang Q, Tang W, She Z, et al. A frustrating problem: accelerated blood clearance of PEGylated solid lipid nanoparticles following subcutaneous injection in rats. Eur J Pharm Biopharm. 2012;81(3):506–13.CrossRefPubMedPubMedCentralGoogle Scholar
  24. 24.
    Ishida T, Ichikawa T, Ichihara M, Sadzuka Y, Kiwada H. Effect of the physicochemical properties of initially injected liposomes on the clearance of subsequently injected PEGylated liposomes in mice. J Control Release. 2004;95(3):403–12.CrossRefPubMedPubMedCentralGoogle Scholar
  25. 25.
    Ishida T, Harada M, Wang XY, Ichihara M, Irimura K, Kiwada H. Accelerated blood clearance of PEGylated liposomes following preceding liposome injection: effects of lipid dose and PEG surface-density and chain length of the first-dose liposomes. J Control Release. 2005;105(3):305–17.CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Peng K-W, TenEyck CJ, Galanis E, Kalli KR, Hartmann LC, Russell SJ. Intraperitoneal therapy of ovarian cancer using an engineered measles virus. Cancer Res. 2002;62(16):4656–62.PubMedGoogle Scholar
  27. 27.
    Di Giorgio A, Naticchioni E, Biacchi D, Sibio S, Accarpio F, Rocco M, et al. Cytoreductive surgery (peritonectomy procedures) combined with hyperthermic intraperitoneal chemotherapy (HIPEC) in the treatment of diffuse peritoneal carcinomatosis from ovarian cancer. Cancer. 2008;113(2):315–25.CrossRefPubMedPubMedCentralGoogle Scholar
  28. 28.
    Gill RS, Al-Adra DP, Nagendran J, Campbell S, Shi X, Haase E, et al. Treatment of gastric cancer with peritoneal carcinomatosis by cytoreductive surgery and HIPEC: a systematic review of survival, mortality, and morbidity. J Surg Oncol. 2011;104(6):692–8.CrossRefPubMedPubMedCentralGoogle Scholar
  29. 29.
    Williamson SK, Johnson GA, Maulhardt HA, Moore KM, McMeekin D, Schulz TK, et al. A phase I study of intraperitoneal nanoparticulate paclitaxel (Nanotax®) in patients with peritoneal malignancies. Cancer Chemother Pharmacol. 2015;75(5):1075–87.CrossRefPubMedPubMedCentralGoogle Scholar
  30. 30.
    Moreno-Aspitia A, Perez EA. Nanoparticle albumin-bound paclitaxel (ABI-007): a newer taxane alternative in breast cancer. Future Oncol. 2005;1(6):755–62.CrossRefPubMedPubMedCentralGoogle Scholar
  31. 31.
    Lu Z, Tsai M, Lu D, Wang J, Wientjes MG, JL-S A. Tumor-penetrating microparticles for intraperitoneal therapy of ovarian cancer. J Pharmacol Exp Ther. 2008;327(3):673–82.CrossRefPubMedPubMedCentralGoogle Scholar
  32. 32.
    Xu S, Fan H, Yin L, Zhang J, Dong A, Deng L, et al. Thermosensitive hydrogel system assembled by PTX-loaded copolymer nanoparticles for sustained intraperitoneal chemotherapy of peritoneal carcinomatosis. Eur J Pharm Biopharm. 2016;104:251–9.CrossRefPubMedPubMedCentralGoogle Scholar
  33. 33.
    Allen TM, Cullis PR. Liposomal drug delivery systems: from concept to clinical applications. Adv Drug Deliv Rev. 2013;65(1):36–48.CrossRefPubMedPubMedCentralGoogle Scholar
  34. 34.
    Liang K, Wang L, Su Y, Liu M, Feng R, Song Y, et al. Comparison among different “revealers” in the study of accelerated blood clearance phenomenon. Eur J Pharm Sci. 2018;114:210–6.CrossRefPubMedPubMedCentralGoogle Scholar
  35. 35.
    Yen-Rei AY, O’Koren EG, Hotten DF, Kan MJ, Kopin D, Nelson ER, et al. A protocol for the comprehensive flow cytometric analysis of immune cells in normal and inflamed murine non-lymphoid tissues. PLoS One. 2016; Scholar
  36. 36.
    Horny H-P, Sotlar K, Valent P. Mastocytosis: immunophenotypical features of the transformed mast cells are unique among hematopoietic cells. Immunol Allergy Clin North Am. 2014;34(2):315–21.CrossRefPubMedPubMedCentralGoogle Scholar
  37. 37.
    Drummond DC, Meyer O, Hong K, Kirpotin DB, Papahadjopoulos D. Optimizing liposomes for delivery of chemotherapeutic agents to solid tumors. Pharmacol Rev. 1999;51(4):691–744.PubMedGoogle Scholar
  38. 38.
    Dobrovolskaia MA, McNeil SE. Handbook of immunological properties of engineered nanomaterials. 3rd ed. World Scientific: CRC Press; 2013.CrossRefGoogle Scholar
  39. 39.
    Wang J, Kubes P. A reservoir of mature cavity macrophages that can rapidly invade visceral organs to affect tissue repair. Cell. 2016;165(3):668–78.CrossRefGoogle Scholar
  40. 40.
    Tewari D, Java JJ, Salani R, Armstrong DK, Markman M, Herzog T, et al. Long-term survival advantage and prognostic factors associated with intraperitoneal chemotherapy treatment in advanced ovarian cancer: a gynecologic oncology group study. J Clin Oncol. 2015;33(13):1460–6.CrossRefPubMedPubMedCentralGoogle Scholar
  41. 41.
    Sadeghi B, Arvieux C, Glehen O, Beaujard AC, Rivoire M, Baulieux J, et al. Peritoneal carcinomatosis from non-gynecologic malignancies. Cancer. 2000;88(2):358–63.CrossRefGoogle Scholar
  42. 42.
    Yonemura Y, Ishibashi H, Hirano M, Mizumoto A, Takeshita K, Noguchi K, et al. Effects of neoadjuvant laparoscopic hyperthermic intraperitoneal chemotherapy and neoadjuvant intraperitoneal/systemic chemotherapy on peritoneal metastases from gastric cancer. Ann Surg Oncol. 2017;24(2):478–85.CrossRefPubMedPubMedCentralGoogle Scholar
  43. 43.
    Gesson-Paute A, Ferron G, Thomas F, de Lara EC, Chatelut E, Querleu D. Pharmacokinetics of oxaliplatin during open versus laparoscopically assisted heated intraoperative intraperitoneal chemotherapy (HIPEC): an experimental study. Ann Surg Oncol. 2008;15(1):339–44.CrossRefPubMedPubMedCentralGoogle Scholar
  44. 44.
    Goéré D, Gras-Chaput N, Aupérin A, Flament C, Mariette C, Glehen O, et al. Treatment of gastric peritoneal carcinomatosis by combining complete surgical resection of lesions and intraperitoneal immunotherapy using catumaxomab. BMC Cancer. 2014;14(1):148–55.CrossRefPubMedPubMedCentralGoogle Scholar
  45. 45.
    Deraco M, Rossi CR, Pennacchioli E, Guadagni S, Somers DC, Santoro N, et al. Cytoreductive surgery followed by intraperitoneal hyperthermic perfusion in the treatment of recurrent epithelial ovarian cancer: a phase II clinical study. Tumori. 2001;87(3):120–6.CrossRefPubMedPubMedCentralGoogle Scholar
  46. 46.
    Beutler B, Milsark I, Cerami A. Passive immunization against cachectin/tumor necrosis factor protects mice from lethal effect of endotoxin. Science. 1985;229(4716):869–71.CrossRefGoogle Scholar
  47. 47.
    Heikenwalder M, Polymenidou M, Junt T, Sigurdson C, Wagner H, Akira S, et al. Lymphoid follicle destruction and immunosuppression after repeated CpG oligodeoxynucleotide administration. Nat Med. 2004;10(2):187–92.CrossRefPubMedPubMedCentralGoogle Scholar
  48. 48.
    MacQueen G, Marshall J, Perdue M, Siegel S, Bienenstock J. Pavlovian conditioning of rat mucosal mast cells to secrete rat mast cell protease II. Science. 1989;243(4887):83–5.CrossRefPubMedPubMedCentralGoogle Scholar
  49. 49.
    Galli SJ, Nakae S, Tsai M. Mast cells in the development of adaptive immune responses. Nat Immunol. 2005;6(2):135–42.CrossRefPubMedPubMedCentralGoogle Scholar
  50. 50.
    Jatana S, Palmer BC, Phelan SJ, DeLouise LA. Immunomodulatory effects of nanoparticles on skin allergy. Sci Rep. 2017;7(1):3979–89.CrossRefPubMedPubMedCentralGoogle Scholar
  51. 51.
    Galli SJ, Grimbaldeston M, Tsai M. Immunomodulatory mast cells: negative, as well as positive, regulators of immunity. Nat Rev Immunol. 2008;8(6):478–86.CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© American Association of Pharmaceutical Scientists 2018

Authors and Affiliations

  1. 1.School of PharmacyShenyang Pharmaceutical UniversityBenxiPeople’s Republic of China

Personalised recommendations