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      A Novel Single Pulsed Electromagnetic Field Stimulates Osteogenesis of Bone Marrow Mesenchymal Stem Cells and Bone Repair

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          Abstract

          Pulsed electromagnetic field (PEMF) has been successfully applied to accelerate fracture repair since 1979. Recent studies suggest that PEMF might be used as a nonoperative treatment for the early stages of osteonecrosis. However, PEMF treatment requires a minimum of ten hours per day for the duration of the treatment. In this study, we modified the protocol of the single-pulsed electromagnetic field (SPEMF) that only requires a 3-minute daily treatment. In the in vitro study, cell proliferation and osteogenic differentiation was evaluated in the hBMSCs. In the in vivo study, new bone formation and revascularization were evaluated in the necrotic bone graft. Results from the in vitro study showed no significant cytotoxic effects on the hBMSCs after 5 days of SPEMF treatment (1 Tesla, 30 pulses per day). hBMSC proliferation was enhanced in the SPEMF-treated groups after 2 and 4 days of treatment. The osteogenic differentiation of hBMSCs was significantly increased in the SPEMF-treated groups after 3–7 days of treatment. Mineralization also increased after 10, 15, 20, and 25 days of treatment in SPEMF-treated groups compared to the control group. The 7-day short-course treatment achieved similar effects on proliferation and osteogenesis as the 25-day treatment. Results from the in vivo study also demonstrated that both the 7-day and 25-day treatments of SPEMF increased callus formation around the necrotic bone and also increased new vessel formation and osteocyte numbers in the grafted necrotic bone at the 2 nd and 4 th weeks after surgery. In conclusion, the newly developed SPEMF accelerates osteogenic differentiation of cultured hBMSCs and enhances bone repair, neo-vascularization, and cell growth in necrotic bone in mice. The potential clinical advantage of the SPEMF is the short daily application and the shorter treatment course. We suggest that SPEMF may be used to treat fractures and the early stages of osteonecrosis.

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          Most cited references 27

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          Osteogenesis in transplants of bone marrow cells.

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            Non-traumatic avascular necrosis of the femoral head.

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              Accelerated growth and prolonged lifespan of adipose tissue-derived human mesenchymal stem cells in a medium using reduced calcium and antioxidants.

              Human mesenchymal stem cells (MSCs) have been isolated from bone marrow and other adult tissues and are potentially useful for tissue engineering. Adipose tissue has several clear advantages as a starting material for harvesting stem cells, as it is abundant and relatively easy to procure. However, existing methods to expand adipose-derived MSCs are less than optimal. Here we describe a new cell culture method that accelerates greatly the growth rate and prolongs the lifespan of adipose MSCs. This was accomplished by using a growth medium with low calcium and supplemented with N-acetyl-L-cysteine and L-ascorbic acid-2-phosphate. Cells produced early in these cultures displayed characteristics similar to those previously reported for multipotential stem cells, including a high frequency of anchorage- independent growth in soft agar, lack of gap junctional intercellular communication in a cell type with serpiginous morphology, and the expression of Oct-4. Furthermore, these cells could readily be induced to differentiate into adipocytes, osteoblasts, and chondrocytes. Thus, modification of growth medium by reduction of calcium and addition of antioxidants greatly enhanced the growth rate and extended the lifespan of adipose-derived multipotential human MSCs.
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                Author and article information

                Affiliations
                [1 ]Orthopaedic Research Center, College of Medicine, Kaohsiung Medical University Hospital, Kaohsiung Medical University, Kaohsiung, Taiwan
                [2 ]Graduate Institute of Medicine, College of Medicine, Kaohsiung Medical University Hospital, Kaohsiung Medical University, Kaohsiung, Taiwan
                [3 ]Department of Orthopaedics, College of Medicine, Kaohsiung Medical University Hospital, Kaohsiung Medical University, Kaohsiung, Taiwan
                [4 ]Department of Orthopaedics, Kaohsiung Medical University Hospital, Kaohsiung Medical University, Kaohsiung, Taiwan
                [5 ]Department of Orthopaedics, Kaohsiung Municipal Hsiao-Kang Hospital, Kaohsiung Medical University, Kaohsiung, Taiwan
                [6 ]Department of Physiology, College of Medicine, Kaohsiung Medical University Hospital, Kaohsiung Medical University, Kaohsiung, Taiwan
                Faculté de médecine de Nantes, France
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                Conceived and designed the experiments: Y-CF C-CL G-JW M-LH. Performed the experiments: Y-CF C-CL C-HC I-CT M-LH. Analyzed the data: Y-CF C-CL J-KC C-HC I-CT G-JW M-LH. Contributed reagents/materials/analysis tools: Y-CF C-CL J-KC C-HC G-JW M-LH. Wrote the paper: Y-CF C-CL I-CT M-LH.

                Contributors
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, USA )
                1932-6203
                2014
                14 March 2014
                : 9
                : 3
                PONE-D-13-39187
                10.1371/journal.pone.0091581
                3954729
                24632682

                This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                Counts
                Pages: 9
                Funding
                The authors have no support or funding to report.
                Categories
                Research Article
                Biology
                Anatomy and Physiology
                Musculoskeletal System
                Bone
                Model Organisms
                Animal Models
                Mouse
                Molecular Cell Biology
                Cell Growth
                Engineering
                Bioengineering
                Human Factors Engineering
                Mechanical Engineering
                Medicine
                Anatomy and Physiology
                Musculoskeletal System
                Bone
                Oncology
                Cancers and Neoplasms
                Bone and Soft Tissue Sarcomas
                Surgery
                Orthopedic Surgery
                Physics
                Electricity
                Electromagnetics

                Uncategorized

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