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Clinically translatable cell tracking and quantification by MRI in cartilage repair using superparamagnetic iron oxides.

van Buul GM, Kotek G, Wielopolski PA, Farrell E, Bos PK, Weinans H, Grohnert AU, Jahr H, Verhaar JA, Krestin GP, van Osch GJ, Bernsen MR - PLoS ONE (2011)

Bottom Line: Efficacy and various safety aspects of SPIO cell labeling were determined using appropriate assays.SPIO labeling appears to be safe without influencing cell behavior.SPIO labeled cells can be visualized in an intra-articular environment and quantified when seeded in cartilage defects.

View Article: PubMed Central - PubMed

Affiliation: Department of Radiology, Erasmus MC, Rotterdam, The Netherlands.

ABSTRACT

Background: Articular cartilage has very limited intrinsic regenerative capacity, making cell-based therapy a tempting approach for cartilage repair. Cell tracking can be a major step towards unraveling and improving the repair process of these therapies. We studied superparamagnetic iron oxides (SPIO) for labeling human bone marrow-derived mesenchymal stem cells (hBMSCs) regarding effectivity, cell viability, long term metabolic cell activity, chondrogenic differentiation and hBMSC secretion profile. We additionally examined the capacity of synovial cells to endocytose SPIO from dead, labeled cells, together with the use of magnetic resonance imaging (MRI) for intra-articular visualization and quantification of SPIO labeled cells.

Methodology/prinicipal findings: Efficacy and various safety aspects of SPIO cell labeling were determined using appropriate assays. Synovial SPIO re-uptake was investigated in vitro by co-labeling cells with SPIO and green fluorescent protein (GFP). MRI experiments were performed on a clinical 3.0T MRI scanner. Two cell-based cartilage repair techniques were mimicked for evaluating MRI traceability of labeled cells: intra-articular cell injection and cell implantation in cartilage defects. Cells were applied ex vivo or in vitro in an intra-articular environment and immediately scanned. SPIO labeling was effective and did not impair any of the studied safety aspects, including hBMSC secretion profile. SPIO from dead, labeled cells could be taken up by synovial cells. Both injected and implanted SPIO-labeled cells could accurately be visualized by MRI in a clinically relevant sized joint model using clinically applied cell doses. Finally, we quantified the amount of labeled cells seeded in cartilage defects using MR-based relaxometry.

Conclusions: SPIO labeling appears to be safe without influencing cell behavior. SPIO labeled cells can be visualized in an intra-articular environment and quantified when seeded in cartilage defects.

Show MeSH
Chondrogenic differentiation of hBMSCs is not affected by SPIO labeling.Collagen II immunohistochemistry and Perl's iron stain demonstrated SPIO labeling to have no effect on pellet size or collagen II production, while iron remained present in differentiated pellets (A–D). Glycosaminoglycan deposition was also unaffected, as can be seen by thionin stain (E–H) and DMB assay (I). Gene expression of collagen II was unaltered by SPIO labeling (J). All analyses were performed after 35 days of differentiation. Insets display low magnification overviews of the entire pellets. Results shown for triplicate samples from two hBMSC donors.
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pone-0017001-g002: Chondrogenic differentiation of hBMSCs is not affected by SPIO labeling.Collagen II immunohistochemistry and Perl's iron stain demonstrated SPIO labeling to have no effect on pellet size or collagen II production, while iron remained present in differentiated pellets (A–D). Glycosaminoglycan deposition was also unaffected, as can be seen by thionin stain (E–H) and DMB assay (I). Gene expression of collagen II was unaltered by SPIO labeling (J). All analyses were performed after 35 days of differentiation. Insets display low magnification overviews of the entire pellets. Results shown for triplicate samples from two hBMSC donors.

Mentions: Chondrogenic differentiation was studied in cell pellets consisting of labeled and unlabeled cells at different ratios (0, 10, 50 and 100% of labeled cells). Differentiation was unaffected in all pellets containing labeled hBMSCs (10–100%) compared to control pellets consisting of unlabeled cells only. All pellets displayed a cartilaginous extracellular matrix rich in GAG and collagen type II (Figure 2A–H). Moreover, quantitative measurements revealed SPIO-labeled hBMSCs to produce comparable amounts of GAG (Figure 2I). In addition, gene expression of collagen type II was unaltered in SPIO-labeled hBMSCs after chondrogenic differentiation (Figure 2J).


Clinically translatable cell tracking and quantification by MRI in cartilage repair using superparamagnetic iron oxides.

van Buul GM, Kotek G, Wielopolski PA, Farrell E, Bos PK, Weinans H, Grohnert AU, Jahr H, Verhaar JA, Krestin GP, van Osch GJ, Bernsen MR - PLoS ONE (2011)

Chondrogenic differentiation of hBMSCs is not affected by SPIO labeling.Collagen II immunohistochemistry and Perl's iron stain demonstrated SPIO labeling to have no effect on pellet size or collagen II production, while iron remained present in differentiated pellets (A–D). Glycosaminoglycan deposition was also unaffected, as can be seen by thionin stain (E–H) and DMB assay (I). Gene expression of collagen II was unaltered by SPIO labeling (J). All analyses were performed after 35 days of differentiation. Insets display low magnification overviews of the entire pellets. Results shown for triplicate samples from two hBMSC donors.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3044153&req=5

pone-0017001-g002: Chondrogenic differentiation of hBMSCs is not affected by SPIO labeling.Collagen II immunohistochemistry and Perl's iron stain demonstrated SPIO labeling to have no effect on pellet size or collagen II production, while iron remained present in differentiated pellets (A–D). Glycosaminoglycan deposition was also unaffected, as can be seen by thionin stain (E–H) and DMB assay (I). Gene expression of collagen II was unaltered by SPIO labeling (J). All analyses were performed after 35 days of differentiation. Insets display low magnification overviews of the entire pellets. Results shown for triplicate samples from two hBMSC donors.
Mentions: Chondrogenic differentiation was studied in cell pellets consisting of labeled and unlabeled cells at different ratios (0, 10, 50 and 100% of labeled cells). Differentiation was unaffected in all pellets containing labeled hBMSCs (10–100%) compared to control pellets consisting of unlabeled cells only. All pellets displayed a cartilaginous extracellular matrix rich in GAG and collagen type II (Figure 2A–H). Moreover, quantitative measurements revealed SPIO-labeled hBMSCs to produce comparable amounts of GAG (Figure 2I). In addition, gene expression of collagen type II was unaltered in SPIO-labeled hBMSCs after chondrogenic differentiation (Figure 2J).

Bottom Line: Efficacy and various safety aspects of SPIO cell labeling were determined using appropriate assays.SPIO labeling appears to be safe without influencing cell behavior.SPIO labeled cells can be visualized in an intra-articular environment and quantified when seeded in cartilage defects.

View Article: PubMed Central - PubMed

Affiliation: Department of Radiology, Erasmus MC, Rotterdam, The Netherlands.

ABSTRACT

Background: Articular cartilage has very limited intrinsic regenerative capacity, making cell-based therapy a tempting approach for cartilage repair. Cell tracking can be a major step towards unraveling and improving the repair process of these therapies. We studied superparamagnetic iron oxides (SPIO) for labeling human bone marrow-derived mesenchymal stem cells (hBMSCs) regarding effectivity, cell viability, long term metabolic cell activity, chondrogenic differentiation and hBMSC secretion profile. We additionally examined the capacity of synovial cells to endocytose SPIO from dead, labeled cells, together with the use of magnetic resonance imaging (MRI) for intra-articular visualization and quantification of SPIO labeled cells.

Methodology/prinicipal findings: Efficacy and various safety aspects of SPIO cell labeling were determined using appropriate assays. Synovial SPIO re-uptake was investigated in vitro by co-labeling cells with SPIO and green fluorescent protein (GFP). MRI experiments were performed on a clinical 3.0T MRI scanner. Two cell-based cartilage repair techniques were mimicked for evaluating MRI traceability of labeled cells: intra-articular cell injection and cell implantation in cartilage defects. Cells were applied ex vivo or in vitro in an intra-articular environment and immediately scanned. SPIO labeling was effective and did not impair any of the studied safety aspects, including hBMSC secretion profile. SPIO from dead, labeled cells could be taken up by synovial cells. Both injected and implanted SPIO-labeled cells could accurately be visualized by MRI in a clinically relevant sized joint model using clinically applied cell doses. Finally, we quantified the amount of labeled cells seeded in cartilage defects using MR-based relaxometry.

Conclusions: SPIO labeling appears to be safe without influencing cell behavior. SPIO labeled cells can be visualized in an intra-articular environment and quantified when seeded in cartilage defects.

Show MeSH