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Hyperosmolar blood–brain barrier opening using intra-arterial injection of hyperosmotic mannitol in mice under real-time MRI guidance

Nature Protocols volume 17, pages 76–94 (2022)Cite this article

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Abstract

The blood–brain barrier (BBB) is the main obstacle to the effective delivery of therapeutic agents to the brain, compromising treatment efficacy for a variety of neurological disorders. Intra-arterial (IA) injection of hyperosmotic mannitol has been used to permeabilize the BBB and improve parenchymal entry of therapeutic agents following IA delivery in preclinical and clinical studies. However, the reproducibility of IA BBB manipulation is low and therapeutic outcomes are variable. We demonstrated that this variability could be highly reduced or eliminated when the procedure of osmotic BBB opening is performed under the guidance of interventional MRI. Studies have reported the utility and applicability of this technique in several species. Here we describe a protocol to open the BBB by IA injection of hyperosmotic mannitol under the guidance of MRI in mice. The procedures (from preoperative preparation to postoperative care) can be completed within ~1.5 h, and the skill level required is on par with the induction of middle cerebral artery occlusion in small animals. This MRI-guided BBB opening technique in mice can be utilized to study the biology of the BBB and improve the delivery of various therapeutic agents to the brain.

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Fig. 1: BBBO under real-time MRI guidance in mice.
Fig. 2: Schematic illustration of BBBO under real-time MRI guidance in large animals and humans.
Fig. 3: Schematic diagram of the catheter assembly.
Fig. 4: Schematic diagram of the preoperative preparation of the mouse.
Fig. 5: Surgical procedures before catheter insertion.
Fig. 6: Schematic diagram of catheter cannulation.
Fig. 7: Mouse MRI setup.
Fig. 8: Use of real-time MRI to ensure an effective infusion rate via IA injection to predict perfusion territory in a mouse brain.
Fig. 9: Variability of cortical involvement during IA infusion of a contrast agent in the mouse brain.
Fig. 10: Prediction of mannitol-induced BBBO territory.
Fig. 11: MRI and histological assessment post-BBBO.

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Data availability

Source data are provided with this paper. The other data that support the findings of this study are available from the corresponding author upon reasonable request.

Code availability

The code used in this study is provided in Supplementary Data 1. We have also deposited the code and a demonstration of image processing at https://github.com/dychuchengyan/ChengyanMRI.

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Acknowledgements

This work was financially supported by 2017-MSCRFF-3942, 2019-MSCRFF-5031, NIH R01NS091110, R01NS102675 and R21NS091599. We thank I.-H. Wu for preparing Fig. 1 and B. Pocta for editorial assistance.

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Authors and Affiliations

  1. Department of Diagnostic Radiology and Nuclear Medicine, University of Maryland School of Medicine, Baltimore, MD, USA

    Chengyan Chu, Anna Jablonska, Yue Gao, Xiaoyan Lan, Yajie Liang, Monica Pearl, Miroslaw Janowski & Piotr Walczak

  2. Dalian Municipal Central Hospital affiliated with Dalian Medical University, Dalian, China

    Chengyan Chu & Shen Li

  3. Russell H. Morgan Department of Radiology and Radiological Science, The Johns Hopkins University School of Medicine, Baltimore, MD, USA

    Wojciech G. Lesniak & Guanshu Liu

  4. Department of Neurology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany

    Tim Magnus

  5. Neurointerventional Radiology, Children’s National Medical Center, Washington, DC, USA

    Monica Pearl

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Contributions

P.W., M.J., M.P., T.M., S.L. and C.C. contributed to conception and design; C.C., A.J., W.J., Y.G. and X.L. conducted the experiments; C.C., Y.G., G.L. and Y.L. analyzed and interpreted the data; C.C. drafted the manuscript, with revision from A.J., Y.G., X.L., Y.L., W.J., G.L., S.L., T.M., M.P., M.J. and P.W.

Corresponding author

Correspondence to Piotr Walczak.

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Competing interests

M.P., M.J. and P.W. are founders and equity holders in Intra-ART. M.J. and P.W. are founders and equity holders in Ti-Com.

Additional information

Peer review information Nature Protocols thanks Mark S. Bolding, Laura M. Vecchio and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Related links

Key references using this protocol

Chu, C. et al. Front. Neurol. 9, 921 (2018): https://doi.org/10.3389/fneur.2018.00921

Lesniak, W. G. et al. J. Nucl. Med. 60, 617–622 (2019): https://doi.org/10.2967/jnumed.118.218792

Janowski, M. et al. J. Cereb. Blood Flow Metab. 36, 569–575 (2016): https://doi.org/10.1177/0271678X15615875

Supplementary information

Supplementary Information

Supplementary Figs. 1–4 and Supplementary Methods.

Reporting Summary

Supplementary Data 1

Matlab code and a demonstration of image processing

Supplementary Video 1

Temporary ligation of ECA and OA cauterization

Supplementary Video 2

Temporary ligation of PPA

Supplementary Video 3

Catheter cannulation

Supplementary Video 4

Brain perfusion of a contrast agent under real-time MRI

Supplementary Video 5

Postoperative procedures

Source data

Source Data Fig. 8

Statistical source data.

Source Data Fig. 9

Statistical source data.

Source Data Fig. 10

Statistical source data.

Source Data Fig. 11

Statistical source data.

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Chu, C., Jablonska, A., Gao, Y. et al. Hyperosmolar blood–brain barrier opening using intra-arterial injection of hyperosmotic mannitol in mice under real-time MRI guidance. Nat Protoc 17, 76–94 (2022). https://doi.org/10.1038/s41596-021-00634-x

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