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Combining acellular nerve allografts with brainderived neurotrophic factor transfected bone marrow mesenchymal stem cells restores sciatic nerve injury better than either intervention alone

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Neural Regeneration Research 2014年第20期9卷 1814-1819页

作者： Yanru Zhang Hui Zhang Gechen Zhang Ka Ka Wenhua Huang School of International Education Zhengzhou University Institute of Clinical Anatomy Southern Medical University Department of Orthopedic Surgery First Affiliated Hospital of Zhengzhou University

In this study, we chemically extracted acellular nerve allografts from bilateral sciatic nerves, and repaired 10-mm sciatic nerve defects in rats using these grafts and brain-derived neurotrophic factor transfected bone marrow mesenchymal stem cells. Experiments were performed in three groups： the acellular nerve allograft bridging group, acellular nerve allograft ＋ bone marrow mesenchymal stem cells group, and the acellular nerve allograft ＋ brain-derived neurotrophic factor transfected bone marrow mesenchyrnal stem cells group. Results showed that at 8 weeks after bridging, sciatic functional index, triceps wet weight recovery rate, myelin thickness, and number of myelinated nerve fibers were significantly changed in the three groups. Variations were the largest in the acellular nerve allograft ＋ brain-derived neurotrophic factor transfected bone marrow mesenchymal stem cells group compared with the other two groups. Experimental findings suggest that chemically extracted acellular nerve allograft combined nerve factor and mesenchymal stem cells can promote the restoration of sciatic nerve defects. The repair effect seen is better than the single application of acellular nerve allograft or acellular nerve allograft combined mesenchymal stem cell transplantation.

关键词： nerve regeneration peripheral nerve regeneration peripheral nerve injury chemicallyextracted acellular nerve brain-derived neurotrophic factor bone marrow mesenchymal stem cells nerve tissue engineering neural regeneration

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Chondroitinase ABC plus bone marrow mesenchymal stem cells for repair of spinal cord injury

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Neural Regeneration Research 2013年第11期8卷 965-974页

作者： Chun Zhang Xijing He Haopeng Li Guoyu Wang Department of Orthopedics Second Hospital of Xi'an Jiaotong University

As chondroitinase ABC can improve the hostile microenvironment and cell transplantation is proven to be effective after spinal cord injury, we hypothesized that their combination would be a more effective treatment option. At 5 days after T8 spinal cord crush injury, rats were injected with bone marrow mesenchymal stem cell suspension or chondroitinase ABC 1 mm from the edge of spinal cord damage zone. Chondroitinase ABC was first injected, and bone marrow mesenchymal stem cell suspension was injected on the next day in the combination group. At 14 days, the mean Basso, Beattie and Bresnahan score of the rats in the combination group was higher than other groups. Hematoxylin-eosin staining showed that the necrotic area was significantly reduced in the combination group compared with other groups. Glial fibrillary acidic protein-chondroitin sulfate proteoglycan double staining showed that the damage zone of astrocytic scars was significantly reduced without the cavity in the combination group. Glial fibrillary acidic protein/growth associated protein-43 double immunostaining revealed that positive fibers traversed the damage zone in the combination group. These results suggest that the combination of chondroitinase ABC and bone marrow mesenchymal stem cell transplantation contributes to the repair of spinal cord injury.

关键词： neural regeneration spinal cord injury stem cells chondroitin sulfate proteoglycans astrocytes glial scar chondroitinase ABC bone marrow mesenchymal stem cells transplantation chemicalbarrier neuroregeneration

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In vitro chondrogenesis of the goat bone marrow mesenchymal stem cells directed by chondrocytes in monolayer and 3-dimetional indirect co-culture system

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Chinese Medical Journal 2011年第19期124卷 3080-3086页

作者： LI Jian-wei GUO Xiao-lei HE Chun-la TUO Yong-hua WANG Zhao WEN Jun JIN Dan Department of Traumatology and Orthopedics Nanfang Hospital Southern Medical University Guangzhou Guangdong 510515 China Department of Biostatistics Southern Medical University Guangzhu Gungdong 510515 China

Background Cartilage injury has a very poor capacity for intrinsic regeneration. The cell-based treatment strategy for the cartilage repair using differentiated bone marrow mesenchymal stem cells （BMSCs） is, however, a promising approach to the chondral repair. This study was aimed to explore the chondrogenic potential of the goat BMSCs in the Transwell co-culture system and the poly-laetide-co-glycolide （PLGA） scaffolds.Methods The BMSCs were isolated from the goat iliac crest while the chondrocytes were obtained from the goat＇s last costal cartilage. In the Transwell co-culture system, the BMSCs co-cultured with chondrocytes were designed as group A,whereas the goat＇s BMSCs induced with the chondrogenic medium were group B. Both groups A and B were the experimental groups, while group C that only contained BMSCs was the control group. In the PLGA scaffolds co-culture system, BMSCs were seeded into the PLGA scaffolds, which were suspended in the 24-well plate, and the control group was established by presence or absence of chondrocytes at the bottom of the 24-well plate. Toluidine blue staining,Alcian blue staining, collagen Ⅱ immunofluoresence, collagen Ⅱ immunochemical staining, collagen Ⅰ, collagen Ⅱ, COL2a Q-PCR and osteopontin Q-PCR were used to examine the chondrogenic conditions as well as the expressions of chondrogenic and osteogenic genes.Results cells isolated from the aspirates of the goat bone marrow proliferated rapidly and gained characteristics of stem cells in Passage 4. However, the differentiations of chondrocytes were not apparent in Passage 3. The results from Toluidine blue staining, collagen Ⅱ immunofluoresence and PCR showed the transformation of BMSCs to chondrocytes in the Transwell co-culture system and PLGA scaffolds. Although the cartilage gene expressions were upgraded in both chondrogenesis group and co-culture system, the osteopontin gene expression, which represents osteogenic level, was also up-regulated.Conclusions The Transwell co-culture system and the PLGA scaffolds co-cultu

关键词： bone marrow mesenchymal stem cells chondrocytes co-culture goat

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bone marrow-derived mesenchymal stem cells modulate autophagy in RAW264.7 macrophages via the phosphoinositide 3-kinase/protein kinase B/heme oxygenase-1 signaling pathway under oxygen-glucose deprivation/restoration conditions

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Chinese Medical Journal 2021年第6期134卷 699-707页

作者： Ning-Fang Wang Chun-Xue Bai Department of Pulmonary Medicine Zhongshan HospitalFudan UniversityShanghai 200032China

Background: Autophagy of alveolar macrophages is a crucial process in ischemia/reperfusion injury-induced acute lung injury (ALI). bone marrow-derived mesenchymal stem cells (BM-MSCs) are multipotent cells with the potential for repairing injured sites and regulating autophagy. This study was to investigate the influence of BM-MSCs on autophagy of macrophages in the oxygen-glucose deprivation/restoration (OGD/R) microenvironment and to explore the potential mechanism.Methods: We established a co-culture system of macrophages (RAW264.7) with BM-MSCs under OGD/R conditionsin vitro. RAW264.7 cells were transfected with recombinant adenovirus (Ad-mCherry-GFP-LC3B) and autophagic status of RAW264.7 cells was observed under a fluorescence microscope. Autophagy-related proteins light chain 3 (LC3)-I, LC3-II, and p62 in RAW264.7 cells were detected by Western blotting. We used microarray expression analysis to identify the differently expressed genes between OGD/R treated macrophages and macrophages co-culture with BM-MSCs. We investigated the gene heme oxygenase-1 (HO-1), which is downstream of the phosphoinositide 3-kinase/protein kinase B (PI3K/Akt) signaling pathway.Results: The ratio of LC3-II/LC3-I of OGD/R treated RAW264.7 cells was increased (1.27 ± 0.20vs. 0.44 ± 0.08,t = 6.67,P cells (0.68 ± 0.14vs. 1.27 ± 0.20,t = 4.12,P < 0.05), up-regulated p62 expression (1.10 ± 0.20vs. 0.77 ± 0.04,t = 2.80,P < 0.05), and up-regulated PI3K (0.54 ± 0.05vs. 0.40 ± 0.06,t = 3.11,P < 0.05) and p-Akt/Akt ratios (0.52 ± 0.05vs. 0.39 ± 0.02,t = 9.13,P < 0.05). A whole-genome microarray assay screened the differentially expressed geneHO-1, which is downstream of the PI3K/Akt signaling pathway, and the alteration ofHO-1 mRNA and pr

关键词： bone marrow mesenchymal stem cells Oxygen-glucose deprivation/restoration Phosphoinositide 3-kinase/protein kinase B signaling pathway Macrophages Autophagy Whole-genome microarray assay

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Intravenous transplantation of bone marrow mesenchymal stem cells promotes neural regeneration after traumatic brain injury

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Neural Regeneration Research 2014年第9期9卷 919-923页

作者： Fatemeh Anbari Mohammad Ali Khalili Ahmad Reza Bahrami Arezoo Khoradmehr Fatemeh Sadeghian Farzaneh Fesahat Ali Nabi Research and Clinical Center for Infertility Shahid Sadoughi University of Medical Sciences Institute of Biotechnology Ferdowsi University of Mashhad

To investigate the supplement of lost nerve cells in rats with traumatic brain injury by intravenous administration of allogenic bone marrow mesenchymal stem cells, this study established a Wistar rat model of traumatic brain injury by weight drop impact acceleration method and administered 3 × 106 rat bone marrow mesenchymal stem cells via the lateral tail vein. At 14 days after cell transplantation, bone marrow mesenchymal stem cells differentiated into neurons and astrocytes in injured rat cerebral cortex and rat neurological function was improved significantly. These findings suggest that intravenously administered bone marrow mesenchymal stem cells can promote nerve cell regeneration in injured cerebral cortex, which supplement the lost nerve cells.

关键词： nerve regeneration bone marrow mesenchymal stem cells traumatic brain injury intravenous administration cell differentiation neurologic function,cerebral cortex rats neural regeneration

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Factors affecting directional migration of bone marrow mesenchymal stem cells to the injured spinal cord

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Neural Regeneration Research 2014年第18期9卷 1688-1695页

作者： Peng Xia Su Pan Jieping Cheng Maoguang Yang Zhiping Qi Tingting Hou Xiaoyu Yang Department of Spine Surgery Orthopedics HospitalSecond HosptalJilin University Department of Endocrinology Second HosptalJilin University

Microtubule-associated protein 1B plays an important role in axon guidance and neuronal migration. In the present study, we sought to discover the mechanisms underlying microtu- bule-associated protein 1B mediation of axon guidance and neuronal migration. We exposed bone marrow mesenchymal stem cells to okadaic acid or N-acetyl-D-erythro-sphingosine （an inhibitor and stimulator, respectively, of protein phosphatase 2A） for 24 hours. The expression of the phosphorylated form of type I microtubule-associated protein 1B in the cells was greater after exposure to okadaic acid and lower after N-acetyl-D-erythro-sphingosine. We then injected the bone marrow mesenchymal stem cells through the ear vein into rabbit models of spinal cord contusion. The migration of bone marrow mesenchymal stem cells towards the injured spinal cord was poorer in cells exposed to okadaic acid- and N-acetyl-D-erythro-sphingosine than in non-treated bone marrow mesenchymal stem cells. Finally, we blocked phosphatidylinosi- tol 3-kinase （PI3K） and extracellular signal-regulated kinase 1/2 （ERK1/2） pathways in rabbit bone marrow mesenchymal stem cells using the inhibitors LY294002 and U0126, respectively. LY294002 resulted in an elevated expression of phosphorylated type I microtubule-associated protein 1B, whereas U0126 caused a reduction in expression. The present data indicate that PI3K and ERKI/2 in bone marrow mesenchymal stem cells modulate the phosphorylation of micro- tubule-associated protein 1B via a cross-signaling network, and affect the migratory efficiency of bone marrow mesenchymal stem cells towards injured spinal cord.

关键词： nerve regeneration bone marrow mesenchymal stem cells spinal cord injury microtubule-associated protein 1 B protein phosphatase 2A cell transplantation phosphorylation signal transduction NSFC grant neural regeneration

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Ectopic osteogenesis and scaffold biodegradation of tissue engineering bone composed of chitosan and osteo-induced bone marrow mesenchymal stem cells in vivo

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Chinese Medical Journal 2014年第2期127卷 322-328页

作者： He Yiqun Dong Youhai Chen Xujun Lin Rongqiang Department of Orthopaedic Surgery the Fifth People's Hospital of Shanghai Fudan University Shanghai 200240 China

Background Chitosan （CS） scaffolds combined with osteogenically induced bone marrow mesenchymal stem cells （BMSCs） have been proved to be promising substitutes for repairing bone defects.Nevertheless,the bone-forming and scaffold-biodegrading processes are seldom studied.This study aimed to determine the osteogenic ability of CS/osteoinduced BMSC composites by observing the bone-forming process and explore the relationship between bone formation and scaffold biodegradation.Methods The CS/osteo-induced BMSC composites （CS＋cells group） and the CS scaffolds （CS group） were,respectively,implanted into SD rat thigh muscles.At 2,4,6,8,and 12 weeks postoperatively,the rat femurs were scanned by CT,and the CT values of the implants were measured and comparatively analyzed.Subsequently,the implants were harvested and stained with hematoxylin and eosin and Masson trichrome,and the percentages of bone area,scaffold area,and collagen area were calculated and compared between the two groups.Results The imaging results showed that the densities of implants of the two groups gradually increased along with time,but the CT values of implants in the CS＋cells group were much higher than in the CS group at the same time point （P ＜0.05）.The histological results showed that the de novo bone and collagen formed in the pores of the scaffolds and gradually increased since 2 weeks postoperation in both groups,and the scaffold gradually degraded along with the boneforming process.However,the comparative analysis results showed that the CS＋cells group gained more de novo bone and collagen formation and had less scaffold than the CS group at the same time point （P ＜0.05）.Conclusion The CS/osteo-induced BMSC composites are excellent bone tissue engineering substitutes,and the scaffold biodegradation is accordant with the bone formation.

关键词： ectopic osteogenesis chitosan biodegradation bone marrow mesenchymal stem cells

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Electrophysiological characteristics of cardiomyocyte-like cells from rat bone marrow derived mesenchymal stem cells by four inductors

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Chinese Medical Journal 2013年第18期126卷 3528-3533页

作者： LIU Bo-wu LU An-lin HOU Jing HUANG Wei HOU Hong HOU Zhao-lei DA Jing AI Shi-yi Department of Cardiology Xijing Hospital Fourth Military Medical University Xi'an Shaanxi 710032 China

Background bone marrow derived mesenchymal stem cel induced by different inductors individually or collectively. In s （BMdMSCs） can differentiate into cardiomyocyte-like cells this study, by inducing BMdMSCs with p53 inhibitor （p-fifty three inhibitor-alpha, PFT-a）, 5-azacytidine （5-AZA）, angiotensin-II （Ang-II） and bone morphogenic protein-2 （BMP-2） we compared the influences of four inductors on the differentiation of rat BMdMSCs into caridomyocyte like-cells. Methods BMdMSCs were collected from the bone marrow of Sprague Dawley rats and after the fourth generation, the purified cells were divided into five groups： 5-AZA （10 μmol/L）, Ang-II （0.1 μmol/L）, PFT-α （20 μmol/L）, BMP-2 （10μg/L） and control. The purity of the BMdMSCs and the cardiac differentiation rates were obtained by flow cytometry. The expressions of cTnT in the BMdMSCs after four weeks of induction were detected by immunofluorescence and the expressions of cTnl and Cx43 detected by Western blotting. The green fluorescent levels reflecting intracellular calcium transient function were determined by laser scanning confocal microscopy. The total potassium current levels of cells were measured on patch clamp. Results All inductors affected to a different degree the differentiation of BMdMSCs into cardiomyocyte-like cells and the expressions of cTnT, cTnl and Cx43 suggesting that the combination of inductors could be an improved method for cardiac regenerative medicine. In addition, the total potassium current level and calcium transient in PFT-a cardiomyocyte-like cells were higher than other groups. Conclusions The cardiac differentiation of BMdMSCs induced by PFT-α, 5-AZA, Ang-II and BMP-2 has been improved at different levels. PFT-a has an advantage of differentiation rate and electrophysiological function over other inductors.

关键词： bone marrow mesenchymal stem cells differentiation cardiomyocyte electrophysiological characteristics

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Citalopram increases the differentiation efficacy of bone marrow mesenchymal stem cells into neuronal-like cells

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Neural Regeneration Research 2014年第8期9卷 845-850页

作者： Javad Verdi Seyed Abdolreza Mortazavi-Tabatabaei Shiva Sharif Hadi Verdi Alireza Shoae-Hassani Department of Applied Cell Sciences School of Advanced Technologies in MedicineTehran University of Medical Sciences Department of stem cells and Tissue Engineering Research Center for Science and Technology in MedicineTehran University of Medical Sciences Department of Tissue Engineering School of Advanced Technologies in MedicineIran University of Medical Sciences

Several studies have demonstrated that selective serotonin reuptake inhibitor antidepressants can promote neuronal cell proliferation and enhance neuroplasticity both in vitro and in vivo. It is hypothesized that citalopram, a selective serotonin reuptake inhibitor, can promote the neuronal differentiation of adult bone marrow mesenchymal stem cells. Citalopram strongly enhanced neuronal characteristics of the cells derived from bone marrow mesenchymal stem cells. The rate of cell death was decreased in citalopram-treated bone marrow mesenchymal stem cells than in control cells in neurobasal medium. In addition, the cumulative population doubling level of the citalopram-treated cells was signiifcantly increased compared to that of control cells. Also BrdU incorporation was elevated in citalopram-treated cells. These ifndings suggest that citalopram can improve the neuronal-like cell differentiation of bone marrow mesenchymal stem cells by increasing cell proliferation and survival while maintaining their neuronal characteristics.

关键词： nerve regeneration citalopram stem cells bone marrow mesenchymal stem cells survival proliferation differentiation neurons neural regeneration

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Electromagnetic Field Change the Expression of Osteogenesis Genes in Murine bone marrow mesenchymal stem cells

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2008年第2期28卷 152-155页

作者：赵东明吴华李峰李锐陶超雄 Department of Orthopedics Tongji HospitalTongji Medical CollegeHuazhong University of Science and Technology Wuhan 430030 China

In order to identify the differentially expressing gene of bone marrow mesenchymal stem cells （MSCs） stimulated by electromagnetic field （EMF） with osteogenesis microarray analysis, the bone marrow MSCs of SD rats were isolated and cultured in vitro. The third-passage cells were stimulated by EMFs and total RNA was extracted, purified and then used for the synthesis of cDNA and cRNA. The cRNA of stimulated group and the control group was hybridized with the rat oligo osteogenesis microarray respectively. The hybridization signals were acquired by using X-ray film after chemiluminescent detection and the data obtained were analyzed by employing the web-based completely integrated GEArray Expression Analysis Suite. RT-PCR was used to identify the target genes： Bmp1, Bmp7, Egf and Egfr. The results showed that 19 differentially expressing genes were found between the stimulated group and the control group. There were 6 up-regulated genes and 13 down-regulated genes in the stimulated group. Semi-quantitative RT-PCR confirmed that the expressions of Bmpl, Bmp7 mRNA of the stimulated group were up-regulated （P〈0.05） and those of Egf, Egfr were down-regulated （P〈0.05）. It was suggested that the gene expression profiles of osteogenesis of the bone marrow MSCs were changed after EMF treatment. It is concluded that the genes are involved in skeletal development, bone mineral metabolism, cell growth and differentiation, cell adhesion etc.

关键词： electromagnetic fields osteogenesis microarray analysis bone marrow mesenchymal stem cells

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