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Fluorescence molecular tomography of DiR-labeled mesenchymal stem cell implants for osteochondral defect repair in rabbit knees

  • Molecular Imaging
  • Published:
European Radiology Aims and scope Submit manuscript

Abstract

Objectives

To assess labelling efficiency of rabbit mesenchymal stem cells (MSCs) using the near-infrared dye 1,1’-dioctadecyl-3,3,3’,3’-tetramethylindotricarbocyanine iodide (DiR) and detection of labelled MSCs for osteochondral defect repair in a rabbit model using fluorescence molecular tomography–X-ray computed tomography (FMT-XCT).

Methods

MSCs were isolated from New Zealand White rabbits and labelled with DiR (1.25-20 μg/mL). Viability and induction of apoptosis were assessed by XTT- and Caspase-3/-7-testing. Chondrogenic potential was evaluated by measurement of glycosaminoglycans. Labelled cells and unlabeled controls (n = 3) underwent FMT-XCT imaging before and after chondrogenic differentiation. Osteochondral defects were created surgically in rabbit knees (n = 6). Unlabeled and labelled MSCs were implanted in fibrin-clots and imaged by FMT-XCT. Statistical analyses were performed using multiple regression models.

Results

DiR-labelling of MSCs resulted in a dose-dependent fluorescence signal on planar images in trans-illumination mode. No significant reduction in viability or induction of apoptosis was detected at concentrations below 10 μg DiR/mL (p > .05); the chondrogenic potential of MSCs was not affected (p > .05). FMT-XCT of labelled MSCs in osteochondral defects showed a significant signal of the transplant (p < .05) with additional high-resolution anatomical information about its osteochondral integration.

Conclusions

FMT-XCT allows for detection of stem cell implantation within osteochondral regeneration processes.

Key Points

• DiR-labelling of MSCs shows no toxic side effects or impairment of chondrogenesis.

• Fluorescence molecular tomography allows for detection of MSCs for osteochondral defect repair.

• FMT-XCT helps to improve evaluation of cell implantation and osteochondral regeneration processes.

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Abbreviations

MSC:

mesenchymal stem cell

DiR:

1,1’-dioctadecyl-3,3,3’,3’-tetramethylindotricarbocyanine iodide

FMT-XCT:

fluorescence molecular tomography–X-ray computed tomography

MR:

magnetic resonance

DAPI:

4′,6-Diamidin-2-phenylindol

GAG:

glycosaminoglycan

TGF-β3:

transforming growth factor beta 3

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Acknowledgments

The scientific guarantor of this publication is Reinhard Meier. The authors of this manuscript declare no relationships with any companies, whose products or services may be related to the subject matter of the article. This study has received funding by the DFG (Deutsche Forschungsgemeinschaft = Germany Research Council): Reinhard Meier acknowledges support from the DFG ME 3718/2-1. Tobias Henning was supported by the DFG HE 4578/3-1. One of the authors (Bernhard Haller, statistician) has significant statistical expertise. Institutional Review Board approval was obtained. Approval from the institutional animal care committee was obtained. Methodology: experimental

Author information

Authors and Affiliations

  1. Department of Orthopaedic Sports Medicine, Klinikum rechts der Isar, Technische Universität München, Munich, Germany

    Markus T. Berninger, Stephan Masius & Andreas B. Imhoff

  2. Department of Trauma and Orthopaedic Surgery, BG Unfallklinik Murnau, Prof.-Küntscher-Strasse 8, 82418, Murnau, Germany

    Markus T. Berninger

  3. Institute for Biological and Medical Imaging, Technische Universität München und Helmholtz Zentrum München, Neuherberg, Germany

    Pouyan Mohajerani, Xiaopeng Ma & Vasilis Ntziachristos

  4. Department of Radiology, Klinikum rechts der Isar, Technische Universität München, Munich, Germany

    Melanie Kimm, Moritz Wildgruber & Reinhard Meier

  5. Department of Clinical Radiology, Universitätsklinikum Münster, Münster, Germany

    Moritz Wildgruber

  6. Institute for Medical Statistics and Epidemiology, Klinikum rechts der Isar, Technische Universität München, Munich, Germany

    Bernhard Haller

  7. Institute for Experimental Oncology and Therapy Research, Klinikum rechts der Isar, Technische Universität München, Munich, Germany

    Martina Anton

  8. Section of Neuroradiology, Uniklinik Köln, Cologne, Germany

    Tobias D. Henning

  9. Section of Neuroradiology, Krankenhaus der Barmherzigen Brüder, Trier, Germany

    Tobias D. Henning

  10. Department of Diagnostic and Interventional Radiology, Universitätsklinikum Ulm, Ulm, Germany

    Reinhard Meier

Authors
  1. Markus T. Berninger
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Corresponding author

Correspondence to Markus T. Berninger.

Additional information

Tobias D. Henning and Reinhard Meier contributed equally to this work.

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Berninger, M.T., Mohajerani, P., Kimm, M. et al. Fluorescence molecular tomography of DiR-labeled mesenchymal stem cell implants for osteochondral defect repair in rabbit knees. Eur Radiol 27, 1105–1113 (2017). https://doi.org/10.1007/s00330-016-4457-5

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  • DOI: https://doi.org/10.1007/s00330-016-4457-5

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