In Vitro Targeted Magnetic Delivery and Tracking of Superparamagnetic Iron Oxide Particles Labeled Stem Cells for Articular Cartilage Defect Repair

To assess a novel cell manipulation technique of tissue engineering with respect to its ability to augment superparamagnetic iron oxide particles （SPIO） labeled mesenchymal stem cells （MSCs） density at a localized cartilage defect site in an in vitro phantom by applying magnetic force. Meanwhile, non-invasive imaging techniques were use to track SPIO-labeled MSCs by magnetic resonance imaging （MRI）. Human bone marrow MSCs were cultured and labeled with SPIO. Fresh degenerated human osteochondral fragments were obtained during total knee arthroplasty and a cartilage defect was created at the center. Then, the osteochondral fragments were attached to the sidewalls of culture flasks filled with phosphate-buffered saline （PBS） to mimic the human joint cavity. The SPIO-labeled MSCs were injected into the culture flasks in the presence of a 0.57 Tesla （T） magnetic force. Before and 90 min after cell targeting, the specimens underwent T2-weighted turbo spin-echo （SET2WI） sequence of 3.0 T MRI. MRI results were compared with histological findings. Macroscopic observation showed that SPIO-labeled MSCs were steered to the target region of cartilage defect. MRI revealed significant changes in signal intensity （P0.01）. HE staining exibited that a great number of MSCs formed a three-dimensional （3D） cell ＂sheet＂ structure at the chondral defect site. It was concluded that 0.57 T magnetic force permits spatial delivery of magnetically labeled MSCs to the target region in vitro. High-field MRI can serve as an very sensitive non-invasive technique for the visualization of SPIO-labeled MSCs.

Specific targeting of angiogenesis in lung cancer with RGD-conjugated ultrasmall superparamagnetic iron oxide particles using a 4.7T magnetic resonance scanner

Background Angiogenesis is an essential step for tumor development and metastasis.The cell adhesion molecule αvβ3 integrin plays an important role in angiogenesis and is a specific marker of tumor angiogenesis.A novel αvβ3 integrintargeted magnetic resonance （MR） imaging contrast agent utilizing Arg-Gly-Asp （RGD） and ultrasmall superparamagnetic iron oxide particles （USPIO） （referred to as RGD-USPIO） was designed and its uptake by endothelial cells was assessed both in vitro and in vivo to evaluate the angiogenic profile of lung cancer.Methods USPIO were coated with-NH3＋ and conjugated with RGD peptides.Prussian blue staining was performed to evaluate the specific uptake of RGD-USPIO by human umbilical vein endothelial cells （HUVECs）.Targeted uptake and subcellular localization of RGD-USPIO in HUVECs were confirmed by transmission electron microscopy （TEM）.The ability of RGD-USPIO to noninvasively assess αvβ3 integrin positive vessels in lung adenocarcinoma A549 tumor xenografts was evaluated with a 4.7T MR scanner.Immunohistochemistry was used to detect αvβ3 integrin expression and vessel distribution in A549 tumor xenografts.Results HUVECs internalized RGD-USPIO significantly more than plain USPIO.The uptake of RGD-USPIO by HUVECs could be competitively inhibited by addition of free RGD.A significant decrease in T2 signal intensity （SI） was observed at the periphery of A549 tumor xenografts at 30 minutes （P 〈0.05） and 2 hours （P 〈0.01） after RGD-USPIO was injected via the tail vein.Angiogenic blood vessels were mainly distributed in the periphery of tumor xenografts with positive αvβ3 integrin expression.Conclusions RGD-USPIO could specifically label αvβ3 integrin and be taken up by HUVECs.This molecular MR imaging contrast agent can specifically evaluate the angiogenic profile of lung cancer using a 4.7T MR scanner.

Detection of neural stem cells function in rats with traumatic brain injury by manganese-enhanced magnetic resonance imaging

Background Previously we had successfully tracked adult human neural stem cells （NSCs） labeled with superparamagnetic iron oxide particles （SPIOs） in host human brain after transplantation in vivo non-invasively by magnetic resonance imaging （MRI）. However, the function of the transplanted NSCs could not be evaluated by the method. In the study, we applied manganese-enhanced MRI （ME-MRI） to detect NSCs function after implantation in brain of rats with traumatic brain injury （TBI） in vivo. Methods Totally 40 TBI rats were randomly divided into 4 groups with 10 rats in each group. In group 1, the TBI rats did not receive NSCs transplantation. MnCI2＂4H20 was intravenously injected, hyperosmolar mannitol was delivered to disrupt rightside blood brain barrier, and its contralateral forepaw was electrically stimulated. In group 2, the TBI rats received NSCs （labeled with SPIO） transplantation, and the ME-MRI procedure was same to group 1. In group 3, the TBI rats received NSCs （labeled with SPIO） transplantation, and the ME-MRI procedure was same to group 1, but diltiazem was introduced during the electrical stimulation period. In group 4, the TBI rats received phosphate buffered saline （PBS） injection, and the ME-MRI procedure was same to group 1. Results Hyperintense signals were detected by ME-MRI in the cortex areas associated with somatosensory in TBI rats of group 2. These signals, which could not be induced in TBI rats of groups 1 and 4, disappeared when diltiazem was introduced in TBI rats of group 3. Conclusion In this initial study, we mapped implanted NSCs activity and its functional participation within local brain area in TBI rats by ME-MRI technique, paving the way for further pre-clinical research.