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Detection and analysis of human serum albumin nanoparticles within phagocytic cells at the resolution of individual live cell or single 3D multicellular spheroid

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Abstract

Since nanoparticles (NPs) have shown great potential in various biomedical applications, live cell response to NPs should be thoroughly explored prior to their in vivo use. In the current study, live cell array (LCA) methodology and unique cell-based assays were used to study the interaction of magnetite (HSA-Mag NP) loaded human serum albumin NPs with phagocytic cells. The LCA enabled cell culturing during HSA-Mag NP accumulation and monolayer or spheroid formation, concomitantly with on-line monitoring of NP internalization. These platforms were also utilized for imaging intercellular links between living cells preloaded with HSA-Mag NP in 2D and 3D resolution. HSA-Mag NP uptake by cells was quantified by imaging, and analyzed using time-resolved measurements. Image analysis of the individual cells in cell populations showed accumulation of HSA-Mag NP by promonocytes and glial cells in a dose- and time-dependent manner. High variability of NP accumulation in individual cells within cell populations, as well as cell subgroups, was evident in both cell types. Following 24 h interaction, uptake of HSA-Mag NP was about 10 times more efficient in glial cells than in activated promonocytes. The presented assays may facilitate detection and analysis of the amount of NPs within individual cells, as well as the rate of NP accumulation and processing in different subsets of living cells. Such data are crucial for estimating predicted drug dosage delivered by NPs, as well as to study possible mechanisms for NP interference with live cells.

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References

  1. Blechinger J, Bauer AT, Torrano AA, Gorzelanny C, Bräuchle C, Schneider SW (2013) Uptake kinetics and nanotoxicity of silica nanoparticles are cell type dependent. Small. doi:10.1002/smll.201301004

  2. Dawidczyk CM, Russell LM, Searson PC (2014) Nanomedicines for cancer therapy: state-of-the-art and limitations to pre-clinical studies that hinder future developments. Front Chem 2:69. doi:10.3389/fchem.2014.00069

  3. Deutsch M, Deutsch A, Shirihai O, Hurevich I, Afrimzon E, Shafran Y, Zurgil N (2006) A novel miniature cell retainer for correlative high-content analysis of individual untethered non-adherent cells. Lab Chip 6:995–1000

  4. Eustaquio T, Leary JF (2012) Single-cell nanotoxicity assays of superparamagnetic iron oxide nanoparticles. In: Reineke J (ed) Nanotoxicity: methods and protocols. Methods Mol Biol 926:69–85. Humana Press

  5. Gao J, Zhang W, Huang P, Zhang B, Zhang X, Xu B (2008) Intracellular spatial control of fluorescent magnetic nanoparticles. J Am Chem Soc 130:3710–3711

  6. Horie M, Kato H, Fujita K, Endoh S, Iwahashi H (2012) In vitro evaluation of cellular response induced by manufactured nanoparticles. Chem Res Toxicol 25(3):605–619

  7. Hoysentruyt LC, Akgos Z, Seyfried TN (2011) Hypothesis: are neoplastic macrophages/microglia present in glioblastoma multiforme? ASN Neuro 3(4):e00064. doi:10.1042/AN20110011

  8. Ibuki Y, Toyooka T (2012) Nanoparticle uptake measured by flow cytometry. In: Reineke J (ed) Nanotoxicity: methods and protocols, Methods Mol Biol 926:157–166. Humana Press, ISBN: 978-1-62703-001-4 (Print) 978-1-62703-002-1 (Online)

  9. Kettiger H, Schipanski A, Wick P, Huwyler J (2013) Engineered nanomaterial uptake and tissue distribution: from cell to organism. Int J Nanomed 8:3255–3269

  10. Kruttwig K, Brueggemann C, Kaijzel E, Vorhagen S, Hilger T, Löwik C, Hoehn M (2009) Development of a three-dimensional in vitro model for longitudinal observation of cell behavior: monitoring by magnetic resonance imaging and optical imaging. Mol Imaging Biol. doi:10.1007/s11307-009-0289-x

  11. Langer K, Balthasar S, Vogel V, Dinauer N, von Briesen H, Schubert D (2003) Optimization of the preparation process for human serum albumin (HSA) nanoparticles. Int J Pharm 257:169–180

  12. Laurent S, Burtea C, Thirifays C, Häfeli UO, Mahmoudi M (2012) Crucial ignored parameters on nanotoxicology: the importance of toxicity assay modifications and “cell vision”. PLoS One 7(1):e29997. doi:10.1371/journal.pone.0029997

  13. Leßig J, Neu B, Glander HJ, Arnhold J, Reibetanz U (2010) Phagocytotic competence of differentiated U937 cells for colloidal drug delivery systems in immune cells. Inflammation 34(2):99–110. doi:10.1007/s10753-010-9213-4

  14. Love SA, Maurer-Jones MA, Thompson JW, Lin YS, Haynes CL (2012) Assessing nanoparticle toxicity. Annu Rev Anal Chem (Palo Alto Calif) 5:181–205

  15. Lukianova-Hleb EY, Ren X, Constantinou PE, Danysh BP, Shenefelt DL, Carson DD, Farach-Carson MC, Kulchitsky VA, Wu X, Wagner DS, Lapotko DO (2012) Improved cellular specificity of plasmonic nanobubbles versus nanoparticles in heterogeneous cell systems. PLoS One 7(4):e34537

  16. Luo Y, Wang C, Hossain M, Qiao Y, Ma L, An J, Su M (2012) Three-dimensional microtissue assay for high-throughput cytotoxicity of nanoparticles. Anal Chem 84(15):6731–6738

  17. Maldonado CR, Salassa L, Gomez-Blanco N, Mareque-Rivas JC (2013) Nano-functionalization of metal complexes for molecular imaging and anticancer therapy. Coord Chem Rev 257(19):2668–2688

  18. Mandarano G, Lodhia J, Eu P, Ferris NJ, Davidson R, Cowell SF (2010) Development and use of iron oxide nanoparticles (Part 2): the application of iron oxide contrast agents in MRI. Biomed Imaging Interv J 6(2):e13. doi:10.2349/biij.6.2.e13

  19. Markovitz-Bishitz Y, Tauber Y, Afrimzon E, Zurgil N, Sobolev M, Shafran Y, Deutsch A, Howitz S, Deutsch M (2010) A polymer microstructure array for the formation, culturing, and high throughput drug screening of breast cancer spheroids. Biomaterials 31(32):8436–8444

  20. Martínez-Vera NP, Schmidt R, Langer K, Zlatev I, Wronski R, Auer E, Havas D, Windisch M, von Briesen H, Wagner S, Stab J, Deutsch M, Pietrzik C, Fazekas F, Ropele S (2014) Tracking of magnetite labeled nanoparticles in the rat brain using MRI. PLoS One 9(3):e92068

  21. Martins S, Costa-Lima S, Carneiro T, Cordeiro-da-Silva A, Souto EB, Ferreira DC (2012) Solid lipid nanoparticles as intracellular drug transporters: an investigation of the uptake mechanism and pathway. Int J Pharm 430(1):216–227

  22. Meister S, Zlatev I, Stab J, Docter D, Baches S, Stauber RH, Deutsch M, Schmidt R, Ropele S, Windisch M, Langer K, Wagner S, von Briesen H, Weggen S, Pietrzik CU (2013) Nanoparticulate flurbiprofen reduces amyloid-β42 generation in an in vitro blood–brain barrier model. Alzheimers Res Ther 5(6):51. doi:10.1186/alzrt225

  23. Sahay G, Alakhova DY, Kabanov AV (2010) Endocytosis of nanomedicines. J Control Release 145(3):182–195

  24. Sebak S, Mirzaei M, Malhotra M, Kulamarva A, Prakash S (2010) Human serum albumin nanoparticles as an efficient noscapine drug delivery system for potential use in breast cancer: preparation and in vitro analysis. Int J Nanomed 5:525–532

  25. Singhal N, Handa U, Bansal C, Mohan H (2011) Neutrophil phagocytosis by tumor cells: a cytological study. Diagn Cytopathol 39:553–555

  26. Sriraman SK, Aryasomayajula B, Torchilin VP (2014) Barriers to drug delivery in solid tumors. Tissue Barriers 2(3):e29528

  27. Wagner S, Zensi A, Wien SL, Tschickardt SE, Maier W, Vogel T, Worek F, Pietrzik CU, Kreuter J, von Briesen H (2012) Uptake mechanism of ApoE-modified nanoparticles on brain capillary endothelial cells as a blood-brain barrier model. PLoS One 7(3):e32568

  28. Weber C, Kreuter J, Langer K (2000) Desolvation process and surface characteristics of HSA-nanoparticles. Int J Pharm 196(2):197–200

  29. Yang Z, Liu ZW, Allaker RP, Reip P, Oxford J, Ahmad Z, Ren G (2010) A review of nanoparticle functionality and toxicity on the central nervous system. J R Soc Interface 7(Suppl 4):S411–S422

  30. Zhu M, Nie G, Meng H, Xia T, Nel A, Zhao Y (2013) Physicochemical properties determine nanomaterial cellular uptake, transport, and fate. Acc Chem Res 46(3):622–631

  31. Zurgil N, Afrimzon E, Deutsch A, Namer Y, Shafran Y, Sobolev M, Tauber Y, Ravid-Hermesh O, Deutsch M (2010) Polymer live-cell array for real-time kinetic imaging of immune cells. Biomaterials 31(18):5022–5029

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Acknowledgments

This study was endowed by the Bequest of Moshe-Shimon and Judith Weisbrodt, by the ERA-NET NEURON Transnational Research Project 2009 and by the Israel Ministry of Health.

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Correspondence to Mordechai Deutsch.

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Afrimzon, E., Zurgil, N., Sobolev, M. et al. Detection and analysis of human serum albumin nanoparticles within phagocytic cells at the resolution of individual live cell or single 3D multicellular spheroid. J Nanopart Res 17, 492 (2015) doi:10.1007/s11051-015-3306-9

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Keywords

  • Human serum albumin NPs
  • Phagocytic cells
  • Single cell/spheroid
  • Image analysis
  • Targeted drug delivery
  • Nanomedicine