The purpose of this study was to (1) compare three different techniques for ferumoxide labeling of mesenchymal stem cells (MSCs), (2) evaluate if ferumoxide labeling allows in vivo tracking of matrix-associated stem cell implants (MASIs) in an animal model, and (3) compare the magnetic resonance imaging (MRI) characteristics of ferumoxide-labeled viable and apoptotic MSCs. MSCs labeled with ferumoxide by simple incubation, protamine transfection, or Lipofectin transfection were evaluated with MRI and histopathology. Ferumoxide-labeled and unlabeled viable and apoptotic MSCs in osteochondral defects of rat knee joints were evaluated over 12 weeks with MRI. Signal to noise ratios (SNRs) of viable and apoptotic labeled MASIs were tested for significant differences using t-tests. A simple incubation labeling protocol demonstrated the best compromise between significant magnetic resonance signal effects and preserved cell viability and potential for immediate clinical translation. Labeled viable and apoptotic MASIs did not show significant differences in SNR. Labeled viable but not apoptotic MSCs demonstrated an increasing area of T2 signal loss over time, which correlated to stem cell proliferation at the transplantation site. Histopathology confirmed successful engraftment of viable MSCs. The engraftment of iron oxide-labeled MASIs by simple incubation can be monitored over several weeks with MRI. Viable and apoptotic MASIs can be distinguished via imaging signs of cell proliferation at the transplantation site.
"Our team developed stem cell labeling protocols that provide a compromise between cellular iron load that allows MR detection (the higher the better) and cellular iron load that ensures preserved stem cell function and differentiation capacity (the lower the better). Optimized cell labeling protocols lead to an unimpaired chondrogenesis of hMSCs when compared to unlabeled controls [15,24]. In general, an iron load of less than 10 picogram per cell has not shown any impairment in chondrogenesis in our experience, although this would have to be confirmed for other cell types. "
[Show abstract][Hide abstract] ABSTRACT: About 43 million individuals in the US currently suffer from disabilities due to arthritis. Cartilage defects are the major source of pain in the affected joints. Current treatments, whilst alleviating some of the clinical symptoms, prove insufficient to cure the underlying irreversible cartilage loss. Stem cells represent a unique source for restoration of cartilage defects. Pre-clinical and clinical trials are currently pursued to investigate the potential of various types of stem cells and stem cell derived chondrocytes to repair arthritic joints. A major challenge with all stem cell-mediated tissue regeneration approaches is death of the transplanted cells with clearance by the immune system. Our current inability to diagnose successful or unsuccessful engraftment of transplanted cells non-invasively in vivo represents a major bottleneck for the development of successful stem cell therapies. A large variety of non-invasive Magnetic Resonance (MR) imaging techniques have been developed over the last decade, which enable sensitive in vivo detection of Matrix Associated Stem Cell Implants (MASI) and early diagnosis of related complications. While initially focused on successfully harvesting cellular MR imaging approaches with easily applicable SuperParamagnetic Iron Oxide Nanoparticles (SPIO), our team began to observe details that will facilitate clinical translation. We therefore started a broader effort to define a comprehensive set of novel, clinically applicable imaging approaches for stem cell transplants in patients. We established immediately clinically applicable nanoparticle labeling techniques for tracking stem cell transplants with MR imaging; we have evaluated the long term MR signal effects of iron oxide nanoparticle labeled MASI in vivo; and we have defined distinct signal characteristics of labeled viable and apoptotic MASI. This review article will provide an overview over these efforts and discuss important implications for clinical translation.
"These studies tend to adopt invasive surgical models of osteoarthritis in either rabbits or rats where cells (MSCs or chondrocytes) are labelled with MNPs, seeded onto scaffolds, such as collagen type I gel  or agarose , and implanted within osteochondral defects. Animals are then monitored over a period ranging from four to twelve weeks while being MR imaged [13,25,36,37]. Particles used in these studies have ranged from very small superparamagnetic iron oxide nanoparticles (11 nm)  to Food and Drug Administration approved particles such as Endorem (150 nm)  and Feraheme (30 nm) . "
[Show abstract][Hide abstract] ABSTRACT: The application of mesenchymal stem cells (MSCs) in treating rheumatoid arthritis (RA) has been made possible by the immunosuppressive and differentiation abilities of these cells. A non-invasive means of assessing cell integration and bio-distribution is fundamental in evaluating the risks and success of this therapy, thereby enabling clinical translation. This paper defines the use of superparamagnetic iron oxide nanoparticles (SPIONs) in conjunction with magnetic resonance imaging (MRI) to image and track MSCs in vivo within a murine model of RA.
Murine MSCs (mMSCs) were isolated, expanded and labelled with SiMAG, a commercially available particle. In vitro MRI visibility thresholds were investigated by labelling mMSCs with SiMAG with concentrations ranging from 0 to 10 μg/ml and resuspending varying cell doses (103 to 5 × 105 cells) in 2 mg/ml collagen prior to MR-imaging. Similarly, in vivo detection thresholds were identified by implanting 3 × 105 mMSCs labelled with 0 to 10 μg/ml SiMAG within the synovial cavity of a mouse and MR-imaging. Upon RA induction, 300,000 mMSCs labelled with SiMAG (10 μg/ml) were implanted via intra-articular injection and joint swelling monitored as an indication of RA development over seven days. Furthermore, the effect of SiMAG on cell viability, proliferation and differentiation was investigated.
A minimum particle concentration of 1 μg/ml (300,000 cells) and cell dose of 100,000 cells (5 and 10 μg/ml) were identified as the in vitro MRI detection threshold. Cell viability, proliferation and differentiation capabilities were not affected, with labelled populations undergoing successful differentiation down osteogenic and adipogenic lineages. A significant decrease (P < 0.01) in joint swelling was measured in groups containing SiMAG-labelled and unlabelled mMSCs implying that the presence of SPIONs does not affect the immunomodulating properties of the cells. In vivo MRI scans demonstrated good contrast and the identification of SiMAG-labelled populations within the synovial joint up to 7 days post implantation. This was further confirmed using histological analysis.
We have been able to monitor and track the migration of stem cell populations within the rheumatic joint in a non-invasive manner. This manuscript goes further to highlight the key characteristics (biocompatible and the ability to create significant contrast at realistic doses within a clinical relevant system) demonstrated by SiMAG that should be incorporated into the design of a new clinically approved tracking agent.
[Show abstract][Hide abstract] ABSTRACT: This study investigated the effect of α-linolenic acid enriched virgin olive, canola and virgin olive oil on body composition, serum lipids, lipid peroxidation and adipose tissue lipoprotein lipase activity in rats. Twenty six male Sprague Dawley rats were fed ad-libitum three diets, containing 40% of dietary calories from fat being either canola (CO), virgin olive (VO), or α-linolenic acid enriched virgin olive oil (LO) for seven weeks. At the end of feeding period, the rats were killed by decapitation and blood samples were analyzed for serum triglycerides (TG), total and HDL-Cholesterol (TC and HDL-C), glucose, insulin, malondialdehyde (MDA) and 4-hydroxy-2(E)-nonenal (4-HNE). Carcasses were analysed for water, fat, and protein content. Individual fat pads were dissected out and weighed. Fat cell size and number, and adipose tissue lipoprotein lipase activity were determined. Results showed that serum triglycerides levels were lower in the CO and LO as compared to VO group. Total plasma cholesterol levels were not different between the CO and LO group but were significantly lower than the VO group. The α-linolenic acid enrichment of VO seems to mimic the effect of CO on blood lipids, and hence could be responsible for reducing triglycerides and total cholesterol, and increasing HDL-C/total cholesterol ratio when compared to VO. Feeding CO, resulted in significantly lower 4-HNE levels as compared to VO and LO. The effects of α-linolenic acid may be explained by its conversion into eicosapentaenoic acid (EPA), which has been previously reported to produce the same effects on plasma lipids. Further studies are needed to elucidate other possible mechanisms of action of α-linolenic acid on the atherogenic factors.
Nutrition Research 04/1999; 49(4-19):601-612. DOI:10.1016/S0271-5317(99)00025-1 · 2.47 Impact Factor
Bingqing Wang, Peng Yu, Tao Li, Yanyan Bian, Xisheng Weng
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