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Characterization of the Inflammatory Response in a Photothrombotic Stroke Model by MRI: Implications for Stem Cell Transplantation

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Abstract

Purpose

The aim of this study was to evaluate the specificity of magnetic resonance imaging (MRI) contrast in a photothrombotic (PT) stroke model with and without engraftment of superparamagnetic iron oxide (SPIO)-labeled stem cells.

Procedures

We monitored animals with PT stroke versus animals with PT stroke and stem cell engraftment by T2/T2*w MRI 4–8 h and 2, 4, 6/7 and 14 days after PT induction. Results were correlated with immunohistochemistry.

Results

T2*w MRI images showed hypointense contrast due to the accumulation of inflammatory cells and corresponding iron accumulation and glial scar formation in the border zone of the lesion, similar as what was observed for SPIO-labeled cells. Histological analysis was thus indispensable to distinguish between labeled transplanted cells and immune cells.

Conclusion

These results raise caution regarding the non-invasive monitoring of SPIO-labeled transplanted stem cells by MRI in models that result in a strong inflammatory response.

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Acknowledgements

We would like to thank Ann Van Santvoort for helping with the MR scans and Ashwini Atre for her technical assistance with cell cultures. We are also thankful to Prof. Raf Sciot for the discussions and guidance regarding the histology. Caroline Vandeputte is a doctoral fellow of the Institute for the Promotion of Innovation through Science and Technology in Flanders (IWT Vlaanderen). Annelies Crabbe is a research assistant of the Flemish Fund for Scientific Research (FWO Vlaanderen). Koen Van Laere is senior clinical research fellow of the FWO Vlaanderen. We gratefully acknowledge the financial support by the European Commission for EC-FP6 network DiMI (LSHB-CT-2005-512146), EC-FP6-STREP-STROKEMAP and the EC-FP7 network ENCITE (2008-201842), and by the Flemish government for the SBO-IWT-060838 BRAINSTIM and the K.U. Leuven Centers of Excellence ‘MoSAIC’ and ‘SCIL’.

Conflict of Interest Disclosure

The authors declare that they have no conflict of interest.

Author information

Authors and Affiliations

  1. Division of Nuclear Medicine, Katholieke Universiteit Leuven, Leuven, Belgium

    Caroline Vandeputte & Koen Van Laere

  2. Molecular Small Animal Imaging Center, Katholieke Universiteit Leuven, Leuven, Belgium

    Caroline Vandeputte, Debby Thomas, Tom Dresselaers, Veerle Baekelandt, Koen Van Laere & Uwe Himmelreich

  3. Laboratory for Neurobiology and Gene Therapy, Molecular Medicine, Katholieke Universiteit Leuven, Leuven, Belgium

    Caroline Vandeputte & Veerle Baekelandt

  4. Biomedical NMR Unit, Katholieke Universiteit Leuven, O&N1–Herestraat 49, bus 505, 3000, Leuven, Belgium

    Debby Thomas, Tom Dresselaers & Uwe Himmelreich

  5. Stem Cell Institute, Katholieke Universiteit Leuven, Leuven, Belgium

    Annelies Crabbe & Catherine Verfaillie

Authors
  1. Caroline Vandeputte
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  2. Debby Thomas
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  3. Tom Dresselaers
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  4. Annelies Crabbe
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  5. Catherine Verfaillie
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  6. Veerle Baekelandt
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  7. Koen Van Laere
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  8. Uwe Himmelreich
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Corresponding author

Correspondence to Uwe Himmelreich.

Additional information

Caroline Vandeputte and Debby Thomas contributed equally.

Significance:

The manuscript describes the accumulation of macrophages and the formation of the microglial scare in a photothrombotic stroke model over time. We have correlated this inflammatory response with the hypointense contrast in T2/T2*w MRI and compared it with the contrast generated by implanted, iron oxide-labeled stem cells. In our opinion, this has major implications for the interpretation of MR images and their specificity in studies that aim to monitor the location/migration of iron oxide-labeled cells non-invasively by MRI.

Electronic Supplementary Material

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Fig. S1

T2 MRI maps showing the development of a photothrombotic lesion over 14 days for one representative rat. MSME images with 10 TE increments of 10 ms and slice thickness of 0.4 mm were used for the calculation of T2 maps. The slices shown above were centered at the lesion based on the high-resolution 3D T2*w FLASH images. a Control image recorded before cortical irradiation. b Two days after lesion induction a hyperintense rim is visible around the lesion. c Four days after irradiation a dark rim appears around the lesion. d Image recorded at 6 days after lesion induction. e Image recorded at 14 days following lesion induction. The dark rim around the lesion is clearly visible. f Cortical T2 relaxation times recorded following photothrombotic lesion induction. The dark line (diamonds) indicates T2 values of the ischemic lesion over time. T2 relaxation values reach their maximum at 2 days following lesion induction and decline gradually. The line with rectangles indicates T2 values of the dark border that appears around the lesion from day 4 after lesion induction. These values also decline in time to values below normal. The line with triangles represents the T2 values in the contralateral hemisphere (control). Data are shown as mean ± SEM (JPEG 95 kb)

Fig. S2

Diffusion-weighted images showing the progression of a photothrombotic lesion for one representative rat. a Control image recorded before lesion induction. b Four to eight hours after lesion induction. The lesion is seen as a hyperintense area within the right cortex indicating restricted diffusion. c Image recorded 2 days after irradiation. A hyperintense halo appears around the lesion. d At 14 days following lesion induction a dark border around the lesion is clearly visible (JPEG 44 kb)

Fig. S3

Immunohistochemical staining for L-chain ferritin of coronal sections in the region surrounding the stroke with corresponding T2*-weighted MR images. a Two days after stroke, b 6 days after stroke, and c 14 days after stroke. Scale bar, 50 μm (c), 100 μm (a, b) (JPEG 84 kb)

Fig. S4

Reduced number of injected cells provide sufficient contrast for the visualization of engrafted cells next to a photothrombotic lesion. Injection of 10% of the number of labeled cells (100,000) 7 days after lesion induction resulted in reduced contrast (a). Cells were located next to the lesion. The arrow indicates a hypointense rim. Four weeks later, the labeled cells were still visible (b). Cells distributed along the corpus callosum but did not reach the lesion area. The arrow indicates the photothrombotic lesion (reduced in size) (JPEG 61 kb)

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Vandeputte, C., Thomas, D., Dresselaers, T. et al. Characterization of the Inflammatory Response in a Photothrombotic Stroke Model by MRI: Implications for Stem Cell Transplantation. Mol Imaging Biol 13, 663–671 (2011). https://doi.org/10.1007/s11307-010-0395-9

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