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Low-frequency pulsed electromagnetic field pretreated bone marrow-derived mesenchymal stem cells promote the regeneration of crush-injured rat mental nerve

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Neural Regeneration Research 2018年第1期13卷 145-153页

作者： NaRi Seo Sung-Ho Lee Kyung won Ju JaeMan Woo BongJu Kim SoungMin Kim jeong won jahng Jong-Ho Lee Department of Oral and Maxillofacial Surgery Graduate School of Dentistry Seoul National University Seoul South Korea Department of Oral and Maxillofacial Surgery Seoul National University Dental Hospital Seoul South Korea Dental Research Institute Seoul National University Seoul South Korea Clinical Translational Research Center for Dental Science (CTRC) Seoul National University Dental Hospital Seoul South Korea

Bone marrow-derived mesenchymal stem cells （BMSCs） have been shown to promote the regeneration of injured peripheral nerves. Pulsed electromagnetic field （PEMF） reportedly promotes the proliferation and neuronal differentiation of BMSCs. Low-frequency PEMF can induce the neuronal differentiation of BMSCs in the absence of nerve growth factors. This study was designed to investigate the effects of low-frequency PEMF pretreatment on the proliferation and function of BMSCs and the effects of low-frequency PEMF pre-treated BMSCs on the regeneration of injured peripheral nerve using in vitro and in vivo experiments.In in vitro experiments, quantitative DNA analysis was performed to determine the proliferation of BMSCs, and reverse transcription-polymerase chain reaction was performed to detect S100 （Schwann cell marker）, glial fibrillary acidic protein （astrocyte marker）, and brain-derived neurotrophic factor and nerve growth factor （neurotrophic factors） mRNA expression. In the in vivo experiments, rat models of crush-injured mental nerve established using clamp method were randomly injected with low-frequency PEMF pretreated BMSCs, unpretreated BMSCs or PBS at the injury site （1 × 106 cells）. DiI-labeled BMSCs injected at the injury site were counted under the fluorescence microscope to determine cell survival. One or two weeks after cell injection, functional recovery of the injured nerve was assessed using the sensory test with von Frey filaments. Two weeks after cell injection, axonal regeneration was evaluated using histomorphometric analysis and retrograde labeling of trigeminal ganglion neurons. In vitro experiment results revealed that low-frequency PEMF pretreated BMSCs proliferated faster and had greater mRNA expression of growth factors than unpretreated BMSCs. In vivo experiment results revealed that compared with injection of unpretreated BMSCs, injection of low-frequency PEMF pretreated BMSCs led to higher myelinated axon count and axon density and more DiI-labeled neurons in the trigeminal ganglia, con

关键词： nerve regeneration mesenchymal stem cells low-frequency pulsed electromagnetic field peripheral nerve injury crush-injured mental nerve

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Human periodontal ligament stem cells repair mental nerve injury

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Neural Regeneration Research 2013年第30期8卷 2827-2837页

作者： Bohan Li Hun-Jong Jung Soung-Min Kim Myung-Jin Kim jeong won jahng Jong-Ho Lee Department of Oral and Maxillofacial Surgery School of Dentistry Binzhou Medical College Department of Occupation and Environment Konkuk Postgraduate Medical School Department of Oral and Maxillofacial Surgery School of Dentistry Seoul National University Dental Research Institute Seoul National University

Human periodontal ligament stem cells are easily accessible and can differentiate into Schwann cells. We hypothesized that human periodontal ligament stem cells can be used as an alternative source for the autologous Schwann cells in promoting the regeneration of injured peripheral nerve. To validate this hypothesis, human periodontal ligament stem cells （1 × 106） were injected into the crush-injured left mental nerve in rats. Simultaneously, autologous Schwann cells （1 × 106） and PBS were also injected as controls. Real-time reverse transcriptase polymerase chain reaction showed that at 5 days after injection, mRNA expression of low affinity nerve growth factor receptor was sig-nificantaly increased in the left trigeminal ganglion of rats with mental nerve injury. Sensory tests, histomorphometric evaluation and retrograde labeling demonstrated that at 2 and 4 weeks after in-jection, sensory function was significantly improved, the numbers of retrograde labeled sensory neurons and myelinated axons were significantly increased, and human periodontal ligament stem cells and autologous Schwann cells exhibited similar therapeutic effects. These findings suggest that transplantation of human periodontal ligament stem cells show a potential value in repair of mental nerve injury.

关键词： neural regeneration peripheral nerve injury stem cells periodontal ligament stem cells mentalnerve Schwann cells cell transplantation sensory nerve neurotrophic factor neuroregeneration

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Human umbilical cord blood-derived mesenchymal stem cells promote regeneration of crush-injured rat sciatic nerves

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Neural Regeneration Research 2012年第26期7卷 2018-2027页

作者： Mi-Ae Sung Hun Jong Jung Jung-Woo Lee Jin-Yong Lee Kang-Mi Pang Sang Bae Yoo Mohammad S. Alrashdan Soung-Min Kim jeong won jahng Jong-Ho Lee Department of Craniofacial Structure & Functional Biology School of Dentistry Seoul National University Seoul Republic of Korea Department of Occupation and Environment Postgraduate College of Medicine Konkuk University Choong-ju Republic of Korea Department of Oral & Maxillofacial Surgery School of Dentistry Seoul National University Seoul Republic of Korea Department of Dentistry Oral and Maxillofacial Surgery Ajou University School of Medicine Suwon Republic of Korea Dental Research Institute Seoul National University Seoul Republic of Korea

Several studies have demonstrated that human umbilical cord blood-derived mesenchymal stem cells can promote neural regeneration following brain injury. However, the therapeutic effects of human umbilical cord blood-derived mesenchymal stem cells in guiding peripheral nerve regeneration remain poorly understood. This study was designed to investigate the effects of human umbilical cord blood-derived mesenchymal stem cells on neural regeneration using a rat sciatic nerve crush injury model. Human umbilical cord blood-derived mesenchymal stem cells （1 ~ 106） or a PBS control were injected into the crush-injured segment of the sciatic nerve. Four weeks after cell injection, brain-derived neurotrophic factor and tyrosine kinase receptor B mRNA expression at the lesion site was increased in comparison to control. Furthermore, sciatic function index, Fluoro Gold-labeled neuron counts and axon density were also significantly increased when compared with control. Our results indicate that human umbilical cord blood-derived mesenchvmal stem cells promote the functinnal r~.RcJv^rv nf P.n I^h-inillr^4 ~r^i~tit, n^r~e

关键词： human umbilical cord blood-derived mesenchymal stem cells sciatic nerve crush injury FluoroGold stem cells peripheral nerve regeneration regeneration neural regeneration

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Peripheral nerve regeneration with cotransplantation of umbilical cord mesenchymal stem cells and Schwann cells in rat sciatic nerve defect

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Neural Regeneration Research 2011年第7期6卷 485-493页

作者： Jin-Yong Lee Hun-Jong Jung Mohammad S. Alrashdan Bohan Li Mi-Ae sung Sang BaeYoo Soung-Min Kim Myung-Jin Kim jeong won jahng Jong-Ho Lee Department of Oral and Maxillofacial Surgery School of Dentistry Seoul National University Seoul Korea Department of Occupation and Environment Konkuk Postgraduate Medical School Choong-Ju Korea Department of Craniofacial Structure and Functional Biology School of Dentistry Seoul National University Seoul Korea Dental Research Institute School of Dentistry Seoul National University Seoul Korea

Previous research has demonstrated that cotransplantation of umbilical cord mesenchymal stem cells （UCMSCs） and Schwann cells （SCs） can repair spinal nerve injury, but few studies have investigated their use in peripheral nerve regeneration. In the present study, we cotransplanted UCMSCs and SCs to repair 5-mm left sciatic nerve defects in rats, and compared the effects of UCMSCs ＋ SCs transplantation with UCMSCs or SCs transplantation alone. After UCMSCs ＋ SCs transplantation, nerve conduction velocity of the left sciatic nerve and gait were both improved. Retrograde tracing analysis demonstrated that the mean count of fluorogold-labeled neurons, as well as the mean axon count and axon density, were significantly greater in the left sciatic nerve after UCMSCs ＋ SCs transplantation, compared with UCMSCs or SCs transplantation alone. Improvements in conduction velocity and increased sheath thickness in the left sciatic nerve were similar after UCMSCs transplantation and UCMSCs ＋ SCs transplantation. These findings suggest that UCMSCs transplantation can promote the repair of sciatic nerve defects to some extent, but that combined UCMSCs ＋ SCs transplantation has a significantly greater regenerative effect.

关键词： Schwann cells human umbilical cord mesenchymal stem cells peripheral nerve re-generation axotomy

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Construction of recombinant human nerve growth factor beta adenovirus and evaluation of its function An in vitro and in vivo study

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Neural Regeneration Research 2010年第16期5卷 1261-1269页

作者： En-Feng Gao Si-Ho Choi Mi-Ae Sung Bo-Han Li Samir Jabaiti Sang Bae Yoo Sung-June Kim Soung-Min Kim jeong won jahng Jong-Ho Lee Department of Stomatology China-Japan Friendship Hospital Beijing 100029 China Center for Stem Cell and Regenerative Medicine Keck School of Medicine University of Sourthern Californina USA Department of Oral and Maxillofacial Surgery & Dental Research Institute Seoul National University Seoul Korea Department of Plastic and Reconstructive Surgery Jordan University Hospital Amman Jordan Department of Electrical Engineering School of Electrical and Computer Science Seoul National University Seoul Korea

Exogenous delivery of nerve growth factor （NGF） promotes neural regeneration. However, the short half-life limits delivery efficacy. Therefore, a long-term, efficient, local delivery tool or scheme is needed. The purpose of this study was to construct a functioning, recombinant, adenoviral vector carrying human NGF-β （hNGF-β） DNA, and to measure expression of the constructed vector in vitro and in vivo. rhNGF-β adenoviral vector containing full-length hNGF-β cDNA was generated by homologous recombination in Escherichia CoIL The rhNGF-β adenovirus was packaged and amplified in human embryonic kidney HEK293 cells. Transformation efficiency, expression and function of rhNGF-β adenovirus for primary Schwann cells, Schwann cell lines, human embryonic kidney HEK 293 cells, CRH myoblasts, and NIH3T3 fibroblasts were evaluated. Subsequently, expression of rhNGF-β adenovirus at the peripheral nerve of rat was also assessed. Recombinant adenoviral vector carrying hNGF-β was successfully constructed and confirmed by restriction endonuclease analysis and DNA sequence analysis. Green fluorescent protein expression was observed in 90% of rhNGF-β adenovirus-infected cells （primary Schwann cells, Schwann cell line, human embryonic kidney HEK 293 cells, CRH myoblasts, and NIH3T3 fibroblasts） compared with non-infected cells. Total mRNA isolated from rhNGF-β adenovirus-infected cells exhibited strong expression. Maximum NGF release was induced by primary cultured Schwann cells at 4 days after infection, which steadily continued for 14 days. PC-12 cells exposed to media conditioned with rhNGF-β adenovirus-infected Schwann cells exhibited increased neurite extension. In vivo experiment revealed that the injected rhNGF-β adenovirus was transfected into the cells at the injected site and promoted expression of NGF, p75NTR and brain derived neurotrophic factor at the sciatic nerve and dorsal root ganglia.

关键词： nerve growth factor Schwann cell peripheral nerve regeneration adenoviral vector HEK293 cells myoblasts fibroblasts

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Transplantation of mesenchymal stem cells from human umbilical cord versus human umbilical cord blood for peripheral nerve regeneration

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Neural Regeneration Research 2010年第11期5卷 838-845页

作者： Kang-Mi Pang Mi-Ae Sung Mohammad S.Alrashdan Sang Bae Yoo Samir Jabaiti Soung-Min Kim Sung-June Kim Myung-Jin Kim jeong won jahng Jong-Ho Lee Department of Oral Maxillofacial Surgery Seoul National University Seoul 110-768 Korea Department of Craniofacial Structure & Functional Biology Seoul National University Seoul 110-768 Korea Dental Research Institute School of Dentistry Seoul National University Seoul 110-768 Korea Department of Plastic and Reconstructive Surgery Faculty of Medicine Jordan University Hospital Amman 11942 Jordan Department of Electrical Engineering School of Electrical and Computer Science Seoul National University Seoul 110-768 Korea

BACKGROUND： Mesenchymal stem cells （MSCs） appear to be a good alternative to Schwann cells in the treatment of peripheral nerve injury. Fetal stem cells, like umbilical cord blood （UCB） and umbilical cord （UC） stem cells, have several advantages over adult stem cells. OBJECTIVE： To assess the effects of UC-derived MSCs （UCMSCs） and UCB-derived MSCs （UCBMSCs） in repair of sciatic nerve defects. DESIGN, TIME AND SETTING： A randomized controlled animal experiment was performed at the laboratory of Department of Oral and Maxillofacial Surgery, Seoul National University Dental Hospital, from July to December 2009. MATERIALS： UCMSCs were provided by the Research Institute of Biotechnology, Dongguk University. UCBMSCs were provided by the Laboratory of Stem Cells and Tumor Biology, College of Veterinary Medicine, Seoul National University. Dulbecco＇s modified Eagle＇s medium （DMEM） was purchased from Gibco-BRL, USA. METHODS： Seven-week-old Sprague-Dawley rats were randomly and evenly divided into three groups： DMEM, UCBMSCs, and UCMSCs. A 10-mm defect in the left sciatic nerve was constructed in all rats. DMEM （15 μL） containing 1×10^6 UCBMSCs or UCMSCs was injected into the gap between nerve stumps, with the surrounding epineurium as a natural conduit. For the DMEM group, simple DMEM was injected. MAIN OUTCOME MEASURES： At 7 weeks after sciatic nerve dissection, dorsal root ganglia neurons were labeled by fluorogold retrograde labeling. At 8 weeks, electrophysiology and histomorphometry were performed. At 2, 4, 6, and 8 weeks after surgery, sciatic nerve function was evaluated using gait analysis. RESULTS： The UCBMSCs group and the UCMSCs group exhibited similar sciatic nerve function and electrophysiological indices, which were better than the DMEM group, as measured by gait analysis （P 〈 0.05）. Fluorogold retrograde labeling of sciatic nerve revealed that the UCBMSCs group demonstrated a higher number of labeled neurons; however, the differences were not significant. Histomorphometric indices were similar in the UCBMS

关键词： peripheral nerve regeneration umbilical cord mesenchymal stem cell umbilical cord blood mesenchymal stem cell axotomy defect stem cells

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