Fluorescence molecular tomography of DiR-labeled mesenchymal stem cell implants for osteochondral defect repair in rabbit knees
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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).
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.
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.
FMT-XCT allows for detection of stem cell implantation within osteochondral regeneration processes.
• 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.
KeywordsMesenchymal stem cells Osteochondral Cell labelling Fluorescence molecular imaging Rabbit
mesenchymal stem cell
fluorescence molecular tomography–X-ray computed tomography
transforming growth factor beta 3
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
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