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TrkA regulates the regenerative capacity of bone marrow stromal stem cells in nerve grafts

Neural Regeneration Research 2019年第10期14卷 1765-1771页

作者： Mei-Ge Zheng Wen-Yuan Sui Zhen-Dan He Yan Liu Yu-Lin Huang Shu-Hua Mu Xin-Zhong Xu Ji-Sen Zhang Jun-Le Qu Jian Zhang Dong Wang Department of Orthopedics The Seventh Hospital of Sun Yat-sen UniversityShenzhenGuangdong ProvinceChina Department of Orthopedics The Second Hospital of Anhui Medical UniversityHefeiAnhui ProvinceChina School of Medicine Shenzhen UniversityShenzhenGuangdong ProvinceChina Department of Scientific Research The Seventh Hospital of Sun Yat-sen UniversityShenzhenGuangdong ProvinceChina Psychology & Social College of Shenzhen University ShenzhenGuangdong ProvinceChina Key Laboratory of Optoelectronic Devices and Systems of Ministry of Education and Guangdong Province College of Optoelectronic EngineeringShenzhen UniversityShenzhenGuangdong ProvinceChina

We previously demonstrated that overexpression of tropomyosin receptor kinase A(TrkA)promotes the survival and Schwann celllike differentiation of bone marrow stromal stem cells in nerve grafts,thereby enhancing the regeneration and functional recovery of the peripheral nerve.In the present study,we investigated the molecular mechanisms underlying the neuroprotective effects of TrkA in bone marrow stromal stem cells seeded into nerve grafts.Bone marrow stromal stem cells from Sprague-Dawley rats were infected with recombinant lentivirus vector expressing rat TrkA,TrkA-shRNA or the respective control.The cells were then seeded into allogeneic rat acellular nerve allografts for bridging a 1-cm right sciatic nerve defect.Then,8 weeks after surgery,hematoxylin and eosin staining showed that compared with the control groups,the cells and fibers in the TrkA overexpressing group were more densely and uniformly arranged,whereas they were relatively sparse and arranged in a disordered manner in the TrkA-shRNA group.Western blot assay showed that compared with the control groups,the TrkA overexpressing group had higher expression of the myelin marker,myelin basic protein and the axonal marker neurofilament 200.The TrkA overexpressing group also had higher levels of various signaling molecules,including TrkA,pTrkA(Tyr490),extracellular signal-regulated kinases 1/2(Erkl/2),pErk1/2(Thr202/Tyr204),and the anti-apoptotic proteins Bcl-2 and Bcl-xL.In contrast,these proteins were downregulated,while the pro-apoptotic factors Bax and Bad were upregulated,in the TrkA-shRNA group.The levels of the TrkA effectors Akt and pAkt(Ser473)were not different among the groups.These results suggest that TrkA enhances the survival and regenerative capacity of bone marrow stromal stem cells through upregulation of the Erk/Bcl-2 pathway.All procedures were approved by the Animal Ethical and Welfare Committee of Shenzhen University,China in December 2014(approval No.AEWC-2014-001219).

关键词： nerve regeneration bone marrow stromal stem cells tropomyosin receptor kinase A receptor lentiviral vector shRNA extracellular signal-regulated protein kinases 1/2 Bcl-2 nerve grafts peripheral nerve regeneration survival neural regeneration

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Protective effect of thodioloside and bone marrow mesenchymal stem cells infected with HIF-1-expressing adenovirus on acute spinal cord injury

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Neural Regeneration Research 2020年第4期15卷 690-696页

作者： Xiao-Qin Ha Bo Yang Huai-Jing Hou Xiao-Ling Cai Wan-Yuan Xiong Xu-Pan Wei Lanzhou University Second Hospital LanzhouGansu ProvinceChina Department of Clinical Laboratory Lanzhou General Hospital of Lanzhou Military Area CommandLanzhouGansu ProvinceChina School of Clinical Medicine Gansu University of Traditional Chinese MedicineLanzhouGansu ProvinceChina School of Life Science and Engineering Lanzhou University of TechnologyLanzhouGansu ProvinceChina

Rhodioloside has been shown to protect cells from hypoxia injury,and bone marrow mesenchymal stem cells have a good effect on tissue repair.To study the effects of rhodioloside and bone marrow mesenchymal stem cells on spinal cord injury,a rat model of spinal cord injury was established using the Infinite Horizons method.After establishing the model,the rats were randomly divided into five groups.Rats in the control group were intragastrically injected with phosphate buffered saline(PBS)(5μL).PBS was injected at 6 equidistant points around 5 mm from the injury site and at a depth of 5 mm.Rats in the rhodioloside group were intragastrically injected with rhodioloside(5 g/kg)and intramuscularly injected with PBS.Rats in the mesenchymal stem cell(MSC)group were intramuscularly injected with PBS and intramuscularly with MSCs(8×10^6/mL in a 50-μL cell suspension).Rats in the Ad-HIF-MSC group were intragastrically injected with PBS and intramuscularly injected with HIF-1 adenovirus-infected MSCs.Rats in the rhodioloside+Ad-HIF-MSC group were intramuscularly injected with MSCs infected with the HIF-1 adenovirus and intragastrically injected with rhodioloside.One week after treatment,exercise recovery was evaluated with a modified combined behavioral score scale.Hematoxylin-eosin staining and Pischingert’s methylene blue staining were used to detect any histological or pathological changes in spinal cord tissue.Levels of adenovirus IX and Sry mRNA were detected by real-time quantitative polymerase chain reaction and used to determine the number of adenovirus and mesenchymal stem cells that were transfected into the spinal cord.Immunohistochemical staining was applied to detect HIF-1 protein levels in the spinal cord.The results showed that:(1)compared with the other groups,the rhodioloside+Ad-HIF-MSC group exhibited the highest combined behavioral score(P<0.05),the most recovered tissue,and the greatest number of neurons,as indicated by Pischingert’s methylene blue staining.(2)Compared with the PBS group,HIF-1 protein ex

关键词： acute spinal cord injury adenovirus adenovirus gene IX bone marrow mesenchymal stem cells combined behavioral score scale HIF-1α nerve regeneration nerve repair Rhodiola rosea Sry

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Collagen-chitosan scaffold impregnated with bone marrow mesenchymal stem cells for treatment of traumatic brain injury

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Neural Regeneration Research 2019年第10期14卷 1780-1786页

作者： Feng Yan Ming Li Hong-Qi Zhang Gui-Lin Li Yang Hua Ying Shen Xun-Ming Ji Chuan-Jie Wu Hong An Ming Ren Department of Neurosurgery Xuanwu HospitalCapital Medical UniversityBeijingChina China-America Institute of Neuroscience Xuanwu HospitalCapital Medical UniversityBeijingChina Department of Ultrasound Xuanwu HospitalCapital Medical UniversityBeijingChina Department of Traditional Chinese Medicine Xuanwu HospitalCapital Medical UniversityBeijingChina Department of Neurology Xuanwu HospitalCapital Medical UniversityBeijingChina

Combinations of biomaterials and cells can effectively target delivery of cells or other therapeutic factors to the brain to rebuild damaged nerve pathways after brain injury.Porous collagen-chitosan scaffolds were prepared by a freeze-drying method based on brain tissue engineering.The scaffolds were impregnated with rat bone marrow mesenchymal stem cells.A traumatic brain injury rat model was established using the 300 g weight free fall impact method.Bone marrow mesenchymal stem cells/collagen-chitosan scaffolds were implanted into the injured brain.Modified neurological severity scores were used to assess the recovery of neurological function.The Morris water maze was employed to determine spatial learning and memory abilities.Hematoxylin-eosin staining was performed to measure pathological changes in brain tissue.Immunohistochemistry was performed for vascular endothelial growth factor and for 5-bromo-2-deoxyuridine(BrdU)/neuron specific enolase and BrdU/glial fibrillary acidic protein.Our results demonstrated that the transplantation of bone marrow mesenchymal stem cells and collagen-chitosan scaffolds to traumatic brain injury rats remarkably reduced modified neurological severity scores,shortened the average latency of the Morris water maze,increased the number of platform crossings,diminished the degeneration of damaged brain tissue,and increased the positive reaction of vascular endothelial growth factor in the transplantation and surrounding areas.At 14 days after transplantation,increased BrdU/glial fibrillary acidic protein expression and decreased BrdU/neuron specific enolase expression were observed in bone marrow mesenchymal stem cells in the injured area.The therapeutic effect of bone marrow mesenchymal stem cells and collagen-chitosan scaffolds was superior to stereotactic injection of bone marrow mesenchymal stem cells alone.To test the biocompatibility and immunogenicity of bone marrow mesenchymal stem cells and collagen-chitosan scaffolds,immunosuppressive cyclosporine was intravenously inject

关键词： nerve regeneration stem cells collagen chitosan scaffolds traumatic brain injury bone marrow mesenchymal stem cells brain tissue engineering neural regeneration

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Pilose antler aqueous extract promotes the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells by stimulating the BMP-2/Smad1, 5/Runx2 signaling pathway

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Chinese Journal of Natural Medicines 2019年第10期17卷 756-767页

作者： REN Cong GONG Wei LI Feng XIE Ming School of Pharmacy L iaoning University of Traditional Chinese MedicineDalian 116600China Affiliated Hospital of Liaoning University of Traditional Chinese Medicine Shenyang 110032China

Peptides from Pilose antler aqueous extract(PAAE) have been shown to stimulate the proliferation and differentiation of bone marrow mesenchymal stem cells(BMSCs). However, the underlying molecular mechanisms are not well understood. Here, PAAE was isolated and purified to explore the molecular mechanisms underlying PAAE’s effects on BMSCs as well as its osteoprotective effects in ovariectomized rats. Our results showed that PAAE promoted proliferation and differentiation of BMSCs to become osteoblasts by enhancing ALP activity and increasing extracellular matrix mineralization. The trabecular microarchitecture of ovariectomized rats was also found to be protected by PAAE. Quantitative reverse transcription-polymerase chain reaction(Quantitative RT-PCR) results suggest that PAAE also increased the expression of osteogenic markers including, alkaline phosphatase(ALP), runt-related transcription factor 2(Runx2), osteocalcin(OCN), bone morphogenetic protein-2(BMP-2), and collagen I(COL-I). Immunoblotting results indicated that PAAE upregulated the levels of BMP-2 and Runx2 and was associated with Smad1/5 phosphorylation. PAAE A at the concentration of 200μg·mL^-1 showed the strongest effect on proliferation and osteogenic differentiation of BMSCs after 48 h. Using matrix-assisted laser desorption/ionization time of flight mass spectrometry(MALDI-TOF MS), we identified the molecular weight of PAAE A and found that it is less than 3000 Da and showed several significant peaks. In conclusion, PAAE activates the BMP-2/Smad1, 5/Runx2 pathway to induce osteoblastic differentiation and mineralization in BMSCs and can inhibit OVX-induced bone loss. These mechanisms are likely responsible for its therapeutic effect on postmenopausal osteoporosis.

关键词： Pilose antler Postmenopausal osteoporosis Bone marrow mesenchymal stem cells BMP-2/Smad1,5/Runx2 signaling pathway

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Efficacy of autologous bone marrow mononuclear cell transplantation therapy in patients with refractory diabetic peripheral neuropathy

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Chinese Medical Journal 2019年第1期132卷 11-16页

作者： Hong Mao Wei Wei Xiu-Li Fu Jing-Jian Dong Xiao-Yu Lyu Ting Jia Yang Tang Shi Zhao Department of Endocrinology The Central Hospital of WuhanTongji Medical CollegeHuazhong University of Science and TechnologyWuhanHubei 430014China Department of Economics Nanyang Technological UniversitySingapore 637332Singapore

Background:Owing to the multifactorial nature of the pathogenesis of diabetic peripheral neuropathy (DPN),conventional drug therapies have not been effective.The application of stem cells transplantation may be useful for the treatment of DPN.This study was designed to assess the safety and therapeutic effects of autologous bone marrow mononuclear cells (BMMNCs) transplantation on the treatment of refractory DPN.Methods:One hundred and sixty-eight patients with refractory DPN were recruited and enrolled in the study.They received intramuscular injection of BMMNCs and followed at 1,3,6,12,18,24,and 36 months after the transplantation.Clinical data,Toronto Clinical Scoring System (TCSS),and nerve conduction studies (NCSs) were compared before and after the transplantation.Results:The signs and symptoms of neuropathy were significantly improved after BMMNCs transplantation.The values of the TCSS scores at 1 month (9.68 ± 2.49 vs.12.55 ± 2.19,P<0.001) and 3 months (8.47 ± 2.39 vs.12.55 ± 2.19,P<0.001) after the treatment reduced significantly compared with the baseline value.This decrement remained persistent until the end of the study.The conduction velocity and action potential and sensory nerves were significantly improved after transplantation (3 and 12 months after the treatment vs.the baseline:motor nerve conduction velocity,40.24 ± 2.80 and 41.00 ± 2.22 m/s vs.38.21 ± 2.28 m/s,P<0.001;sensory nerve conduction velocity,36.96 ± 2.26 and 39.15 ± 2.61 m/s vs.40.41 ± 2.22 m/s,P<0.001;compound muscle action potential,4.67 ± 1.05 and 5.50 ± 1.20 μV vs.5.68 ± 1.08 μV,P<0.001;sensory nerve action potential,4.29 ± 0.99 and 5.14 ± 1.26 μV vs.5.41 ± 1.14 μV,P<0.001).No adverse event associated with the treatment was observed during the follow-up period.Conclusions:Autologous transplantation of BMMNCs may be an effective and promising therapeutic strategy for the treatment of refractory DPN.

关键词： Bone marrow mononuclear cells diabetic peripheral neuropathy autologous transplantation Toronto clinical scoring system nerve conduction

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Upregulation of UBAP2L in Bone marrow Mesenchymal Stem Cells Promotes Functional Recovery in Rats with Spinal Cord Injury

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Current Medical Science 2018年第6期38卷 1081-1089页

作者： Guan-lin LIN Huan WANG Jun DAI Xiao LI Ming GUAN Qing DING Huai-xi WANG Huang FANG Department of Orthopaedics Tongji HospitalTongji Medical CollegeHuazhong University of Science and TechnologyWuhan 430030China

Post-translational modifications of cellular proteins with ubiquitin or ubiquitin-like proteins regulate many cellular processes,such as cell proliferation,differentiation,apoptosis, signal transduction,intercellular immune recognition,inflammatory response,stress response,and DNA repair.Nice4/UBAP2L is an important member in the family of ubiquitin-like proteins,and its biological function remains unknown.This study aimed to investigate the effect of UBAP2L on spinal cord injury (SCI).At first,rat bone marrow mesenchymal stem cells (BMSCs)were infected with adeno-associated virus to induce over-expression of Nice4.Subsequently,the infected BMSCs were transplanted into rats suffering from semi-sectioned SCI.The results showed that the over-expression of Nice4 significantly promoted the proliferation and differentiation of BMSCs. In addition,the transplantation of infected BMSCs into the injured area of SCI rats improved the function repair of SCI.Importantly,the immunohistochemical and hematoxylin-eosin staining and RT-PCR results showed that the number of neuronal cells,oligodendrocytes,and astrocytes was significantly increased in the injured area,along with significantly upregulated expression ofcyclin D1 and p38 mitogen-activated protein kinase (MAPK).Meanwhile,the expression of caspase 3 protein was significantly down-regulated.In conclusion,the over-expression of Nice4 gene can promote the functional recovery in SCI rats by promoting cell proliferation and inhibiting apoptosis. The results of this study indicate an alternative option for the clinical treatment of SCI.

关键词： bone marrow mesenchymal stem cells spinal cord injury ubiquitin-associated protein 2-like apoptosis proliferation

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Transplantation of bone marrow-derived endothelial progenitor cells and hepatocyte stem cells from liver fibrosis rats ameliorates liver fibrosis

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World Journal of Gastroenterology 2018年第2期24卷 237-247页

作者： Ling Lan Ran Liu Ling-Yun Qin Peng Cheng Bo-Wei Liu Bing-Yong Zhang Song-Ze Ding Xiu-Ling Li Department of Gastroenterology and Hepatology the People's Hospital of Zhengzhou University(the Henan Provincial People’s Hospital) Department of Oncology Henan Provincial Rongjun Hospital Department of Gastroenterology and Hepatology the Children’s Hospital of Zhengzhou Intensive Care Unit the Second Affiliated Hospitalof Luohe Medical College

AIM To explore the effectiveness for treating liver fibrosisby combined transplantation of bone marrow-derived endothelial progenitor cells(BM-EPCs) and bone marrow-derived hepatocyte stem cells(BDHSCs) from the liver fibrosis environment.METHODS The liver fibrosis rat models were induced with carbon tetrachloride injections for 6 wk. BM-EPCs from rats with liver fibrosis were obtained by different rates of adherence and culture induction. BDHSCs from rats with liver fibrosis were isolated by magnetic bead cell sorting. Tracing analysis was conducted by labeling EPCs with PKH26 in vitro to show EPC location in the liver. Finally, BM-EPCs and/or BDHSCs transplantation into rats with liver fibrosis were performed to evaluate the effectiveness of BM-EPCs and/or BDHSCs on liver fibrosis.RESULTS Normal functional BM-EPCs from liver fibrosis rats were successfully obtained. The co-expression level of CD133 and VEGFR2 was 63.9% ± 2.15%. Transplanted BM-EPCs were located primarily in/near hepatic sinusoids. The combined transplantation of BM-EPCs and BDHSCs promoted hepatic neovascularization, liver regeneration and liver function, and decreased collagen formation and liver fibrosis degree. The VEGF levels were increased in the BM-EPCs(707.10 ± 54.32) and BM-EPCs/BDHSCs group(615.42 ± 42.96), compared with those in the model group and BDHSCs group(P 0.05) compared with those in the BDHSC(AST, TBIL and PT, P < 0.05) and BM-EPCs(TBIL and PT, P < 0.05) groups. Transplantation of BM-EPCs/BDHSCs combination significantly reduced the degree of liver fibrosis(staging score of 1.75 ± 0.25 vs BDHSCs 2.88 ± 0.23 or BMEPCs 2.75 ± 0.16, P < 0.05).CONCLUSION The co

关键词： Bone marrow Endothelial progenitor cells Liver stem cell Cell transplantation Liver fibrosis

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Effects of various antimicrobial agents on multi-directional differentiation potential of bone marrow-derived mesenchymal stem cells

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World Journal of Stem Cells 2019年第6期11卷 322-336页

作者： Hui Li Bing Yue Department of Bone and Joint Surgery Renji Hospital Shanghai Jiaotong University School of Medicine

Antimicrobial drugs of several classes play an important role in the treatment of bone and joint infections. In addition to fighting pathogenic microorganisms, the effects of drugs on local tissues and cells are also related to the course and prognosis of bone and joint infections. The multi-directional differentiation potential of bone marrow-derived mesenchymal stem cells (MSCs) is essential for tissue repair after local injury, which is directly related to the recovery of bone, cartilage, and medullary adipose tissue. Our previous studies and the literature indicate that certain antimicrobial agents can regulate the differentiation potential of bone marrow-derived MSCs. Here, in order to systematically analyze the effects of various antimicrobial drugs on local tissue regeneration, we comprehensively review the studies on the effects of these drugs on MSC differentiation, and classify them according to the three differentiation directions (osteogenesis, chondrogenesis, and adipogenesis). Our review demonstrates the specific effects of different antimicrobial agents on bone marrow-derived MSCs and the range of concentrations at which they work, and provides a basis for drug selection at different sites of infection.

关键词： Antimicrobial agents Bone marrow mesenchymal stem cells Osteogenesis Chondrogenesis Adipogenesis

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Bone marrow microenvironment: The guardian of leukemia stem cells

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World Journal of Stem Cells 2019年第8期11卷 476-490页

作者： Mohammad Houshmand Teresa Mortera Blanco Paola Circosta Narjes Yazdi Alireza Kazemi Giuseppe Saglio Mahin Nikougoftar Zarif Department of Clinical and Biological Sciences University of Turin Turin 10126 Italy Center for Hematology and Regenerative Medicine Karolinska Institutet Department of Medicine Karolinska University Hospital Huddinge Stockholm 14183 Sweden Department of Molecular Genetics Tehran Medical Branch Islamic Azad University Tehran *** Iran Department of Hematology and Blood Banking School of Allied Medical Sciences Shahid Beheshti University of Medical Sciences Tehran *** Iran Blood Transfusion Research Center High Institute for Research and Education in Transfusion Medicine Tehran 146651157 Iran

Bone marrow microenvironment (BMM) is the main sanctuary of leukemic stem cells (LSCs) and protects these cells against conventional therapies. However, it may open up an opportunity to target LSCs by breaking the close connection between LSCs and the BMM. The elimination of LSCs is of high importance, since they follow cancer stem cell theory as a part of this population. Based on cancer stem cell theory, a cell with stem cell-like features stands at the apex of the hierarchy and produces a heterogeneous population and governs the disease. Secretion of cytokines, chemokines, and extracellular vesicles, whether through autocrine or paracrine mechanisms by activation of downstream signaling pathways in LSCs, favors their persistence and makes the BMM less hospitable for normal stem cells. While all details about the interactions of the BMM and LSCs remain to be elucidated, some clinical trials have been designed to limit these reciprocal interactions to cure leukemia more effectively. In this review, we focus on chronic myeloid leukemia and acute myeloid leukemia LSCs and their milieu in the bone marrow, how to segregate them from the normal compartment, and finally the possible ways to eliminate these cells.

关键词： Bone marrow microenvironment Bone marrow niche Leukemic stem cell Chronic myeloid leukemia Acute myeloid leukemia Target therapy

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Bone marrow mesenchymal stem cells combined with Sox2 increase the functional recovery in rat with traumatic brain injury

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Chinese Neurosurgical Journal 2019年第2期5卷 85-91页

作者： Qiang Hao Jian Zheng Yue Hu Hao Wang Department of Neurosurgery Beijing Tiantan HospitalCapital Medical UniversityNo.119 South 4th Ring West RoadFengtai DistrictBeijing 100070China Department of Cardiopulmonary Function Examination Shanxi Provincial Cancer HospitalEmployee Xincun No.3Xing Hua Ling DistrictTaiyuan 030000China Basic Medical Science Department Capital Medical UniversityYou An Men Wai StreetXi Tou Tiao 10#Fengtai DistrictBeijing 100069China

Background:About 10 million individuals suffer from traumatic brain injury (TBI) each year in the world,which is one of the most serious neurological disorders.The morbidity of TBl is 55.5~6.1/100,000 in China,which takes more costing in the therapy,and the outcome of that is not well.Therefore,we expect to find new methods to treat TBI and improve the outcomes of TBI.In the previous studies reviewed,we found that stem cell transplantation may hold promising potential for modifying motor dysfunction induced by TBI.Methods:Twenty-six adult SD rats were involved in our study.Two adult SD rats were used as donors of bone marrow stem cells (BMSCs),and the other adult SD rats were divided into four groups randomly,which were used to establish the TBI models.BMSCs were transduced with lentivirous-Sox2,and we try to examine the effects of Sox2 on the differentiation of BMSCs.Establishment of rat TBI model:Rats were anesthetized using pentobarbital sodium (at a concentration of 1.5% and a dose of 40 mg/kg) and fixed under the stereotaxic device.A 1.0-cm craniotomy was performed lateral to the sagittal suture.The skullcap was carefully removed,and rats were then subjected to TBI using a controlled cortical injury instrument.A standardized parietal contusion was performed using a 20-mg steel rod with a diameter of 4 mm,which dropped from a height of 30 cm.After injury,the incision was sutured,and rats were carefully observed and nursed.Treatments:Seven days after TBI,rats were divided into four groups and were transplanted with BMSC-Sox2,single BMSC,single Lentivirus-Sox2,and PBS into injured brain,respectively.The motor function was tested using the neurological severity score (NSS).Results:We found that the ectopic expression of Sox2 enhanced BMSCs to differentiate into neurons.Seven days after TBI,the rats were treated with BMSC-Sox2,BMSC,Sox2,and PBS.Results showed that NSS were 3.352 ± 0.398 in the BMSC-Sox2 group,4.013 ± 0.495 in the BMSC group,4.968 ± 0.293 in the Sox2 group,and 6.257 ± 0.361 in the PBS group,sugges

关键词： Traumatic brain injury Bone marrow stem cell Sox2 gene Transplantation

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