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A Novel Modality for Functional Imaging in Acute Intervertebral Disk Herniation via Tracking Leukocyte Infiltration

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Abstract

Purpose

Inflammation plays a key role in the progression of intervertebral disk (IVD) herniation and associated low back pain. However, real-time spatial diagnosis of inflammation associated with acute disk herniation has not been investigated. We sought to detect local neutrophil and macrophage infiltration near disk herniation via the formyl peptide receptor 1 (FPR1)-mediated molecular imaging in a disk puncture mouse model to elucidate pathophysiological process of disk herniation.

Procedures

Disk herniation was induced in mouse with an established needle puncture procedure. Degenerative change of disk and infiltration of neutrophils and macrophages were detected with Safranin-O, hematoxylin and eosin (H&E), and immunohistochemical staining after injury. FPR1-specific imaging probes cFLFLF-PEG-Cy7 and [99mTc]HYNIC-PEG-cFLFLF were administered systemically to sham and disk injury mice. Leukocyte infiltration was tracked by in vivo near-infrared fluorescence (NIRF) and single-photon emission tomography (SPECT) imaging. The peptide-receptor binding specificity was further investigated with FPR1−/− mice via ex vivo NIRF scan and in vitro binding assays.

Results

Safranin-O staining exhibited disorganized disk structure and loss of proteoglycan after puncture. Massive inflammatory cells were observed in the anterior region of punctured annulus in the injury group. The majority of neutrophils were detected at 1 through 3 days, while infiltration of macrophages appeared the most at 7 days after injury. NIRF and SPECT images revealed preferential accumulation of cFLFLF probes in herniation site in wild-type mice but not in FPR1−/− mice. Binding of the cFLFLF peptide to FPR1 was also observed in RAW 267.4 cells and macrophages isolated from wild-type mice, whereas much less signal was observed in macrophages from FPR1−/− mice. The presence of macrophage infiltration was also detected in human-herniated disk samples by immunohistochemistry.

Conclusion

For the first time, leukocyte infiltration around acute disk herniation site was detected directly and non-invasively in a timely fashion using FPR1-targeted molecular imaging modalities. Such functional imaging of disk herniation via infiltrated leukocytes would advance the understanding of etiology and facilitate drug delivery and treatment monitoring of disk herniation.

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Acknowledgements

The current study was funded by NIH grants NIAMS R21AR057512 and RO1AR064792.

Author information

Author notes

  1. Yi Zhang

    Present address: Biomedical Imaging Research Institute, Cedars-Sinai Medical Center, 8700 Beverly Blvd. Davis Building, Rm G140, Los Angeles, CA, 90048, USA

Authors and Affiliations

  1. Department of Orthopaedic Surgery, University of Virginia, Rm B051, Cobb Hall, 135 Hospital Dr., Charlottesville, VA, 22908, USA

    Li Xiao, Mengmeng Ding, Adam Shimer, Francis Shen, Li Jin & Xudong Li

  2. Department of Radiology and Biomedical Imaging, University of Virginia, Charlottesville, VA, 22908, USA

    Yi Zhang, Mahendra Chordia & Dongfeng Pan

  3. Department of Cardiovascular Medicine, University of Virginia, Charlottesville, 22908, USA

    David Glover

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  1. Li Xiao
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  2. Mengmeng Ding
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  9. Li Jin
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  10. Xudong Li
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Correspondence to Xudong Li.

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The authors declare that they have no conflict of interest.

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Xiao, L., Ding, M., Zhang, Y. et al. A Novel Modality for Functional Imaging in Acute Intervertebral Disk Herniation via Tracking Leukocyte Infiltration. Mol Imaging Biol 19, 703–713 (2017). https://doi.org/10.1007/s11307-016-1038-6

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  • DOI: https://doi.org/10.1007/s11307-016-1038-6

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