Abstract

Here we describe a blood-cleansing device for sepsis therapy inspired by the spleen, which can continuously remove pathogens and toxins from blood without first identifying the infectious agent. Blood flowing from an infected individual is mixed with magnetic nanobeads coated with an engineered human opsonin—mannose-binding lectin (MBL)—that captures a broad range of pathogens and toxins without activating complement factors or coagulation. Magnets pull the opsonin-bound pathogens and toxins from the blood; the cleansed blood is then returned back to the individual. The biospleen efficiently removes multiple Gram-negative and Gram-positive bacteria, fungi and endotoxins from whole human blood flowing through a single biospleen unit at up to 1.25 liters per h in vitro. In rats infected with Staphylococcus aureus or Escherichia coli, the biospleen cleared >90% of bacteria from blood, reduced pathogen and immune cell infiltration in multiple organs and decreased inflammatory cytokine levels. In a model of endotoxemic shock, the biospleen increased survival rates after a 5-h treatment.

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Acknowledgements

This work was supported by Defense Advanced Research Projects Agency grant N66001-11-1-4180 and contract HR0011-13-C-0025, Department of Defense/Center for Integration of Medicine and Innovative Technology and the Wyss Institute for Biologically Inspired Engineering at Harvard University. We thank M. Montoya-Zavala and D. Breslau for micromachining of the blood-cleansing microdevice and technical support; P. Snell and J. Tomolonis for microbiology assistance; A. Schulte for assistance in biospleen experiments; J. Weaver for help with scanning electron microscopy; R. Betensky for statistical analysis assistance; and A. Onderdonk, M. Puder, A. Nedder and their teams for assistance in developing the rat cecal contents sepsis model. We thank J. Fiering and his team for helpful discussions during the early phase of this project. J.H.K. is a recipient of a Wyss Technology Development Fellowship from the Wyss Institute and a professional development postdoctoral award from Harvard University. Scanning electron microscopy images were obtained at the Center for Nanoscale Systems at Harvard University, supportedby the National Science Foundation under award no. ECS-0335765.

Author information

Author notes

    • Martin Rottman

    Current address: Université de Versailles St-Quentin en Yvelines, Versailles, France and Service de Microbiologie, Hôpitaux Universitaires Île de France Ouest, AP-HP, Garches, France.

    • Joo H Kang
    •  & Michael Super

    These authors contributed equally to this work.

Affiliations

  1. Wyss Institute for Biologically Inspired Engineering, Boston, Massachusetts, USA.

    • Joo H Kang
    • , Michael Super
    • , Chong Wing Yung
    • , Ryan M Cooper
    • , Karel Domansky
    • , Amanda R Graveline
    • , Julia B Berthet
    • , Mark J Cartwright
    • , Alexander L Watters
    • , Martin Rottman
    • , Anna Waterhouse
    • , Nazita Gamini
    • , Melissa J Rodas
    • , Anxhela Kole
    • , Thomas M Valentin
    • , Alexander Diaz
    •  & Donald E Ingber
  2. Vascular Biology Program, Boston Children's Hospital and Harvard Medical School, Boston, Massachusetts, USA.

    • Joo H Kang
    • , Chong Wing Yung
    • , Ryan M Cooper
    • , Tadanori Mammoto
    • , Heather Tobin
    • , Akiko Mammoto
    • , Amanda Jiang
    •  & Donald E Ingber
  3. Harvard School of Engineering and Applied Sciences, Cambridge, Massachusetts, USA.

    • Joo H Kang
    •  & Donald E Ingber
  4. Harvard-MIT Health Sciences and Technology Graduate Program, Cambridge, Massachusetts, USA.

    • Ryan M Cooper
  5. Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts, USA.

    • Kazue Takahashi

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Contributions

J.H.K. designed and performed blood-cleansing experiments with assistance from R.M.C., J.B.B., N.G., A.R.G., A.D. and A.W., and analyzed the data and prepared the manuscript. K.D. contributed to the design and integration of the biospleen device. J.H.K., C.W.Y., A.R.G. and H.T. established the rat sepsis models, and J.H.K. and A.R.G. conducted animal studies with help from T.M., A.M. and A.J.; C.W.Y. designed the prototype biospleen device and obtained preliminary data. M.S. and A.L.W. designed, engineered and produced FcMBL with assistance from M.J.R., J.B.B. and A.K.; M.S., M.J.C. and M.R. performed blood analysis for quantitating LPS levels with help from N.G. and helped establish an endotoxemia model in rats. T.M.V. fabricated devices, performed scanning electron microscopy and assisted with conducting studies. K.T. performed experiments to characterize FcMBL versus native MBL. D.E.I. led efforts to design the device and the opsonin, assisted in data analysis and helped write the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Donald E Ingber.

Supplementary information

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    Supplementary Text and Figures

    Supplementary Figures 1–5, Supplementary Table 1

Videos

  1. 1.

    The fabrication and the magnetic separation principle of the biospleen:

    Schematic drawing and microscopic video showing how the biospleen device is fabricated and how the magnetically opsonized pathogens are separated from the blood channel under flow. Because it is difficult to observe the cell movement across the blood channel in the biospleen device, we demonstrated this in a microfluidic device fabricated from optically clear poly(dimethylsiloxane) (PDMS). To mimic pathogens captured by the magnetic opsonins, fluorescent magnetic particles (8 μm, 1.1g ml–1, UMC4F, Bang Laboratories, Inc., IN, USA) were spiked into human banked blood (1ml) and flowed at 10 μl min–1.

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DOI

https://doi.org/10.1038/nm.3640

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