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Three-dimensional bioprinting sodium alginate/gelatin scaffold combined with neural stem cells and oligodendrocytes markedly promoting nerve regeneration after spinal cord injury

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Regenerative Biomaterials 2022年第1期9卷 522-532页

作者： Shuo Liu Hui Yang Dong Chen Yuanyuan Xie ChenXu Tai Liudi Wang Peng Wang Bin Wang Clinical stem cell Center Nanjing Drum Tower Hospitalthe Affiliated Hospital of Nanjing University Medical SchoolNanjing 210000Jiangsu ProvinceChina State Key Laboratory of Pharmaceutical Biotechnology Department of Sports Medicine and Adult Reconstructive SurgeryNanjing Drum Tower HospitalThe Affiliated Hospital of Nanjing University Medical SchoolNanjing 210000Jiangsu ProvinceChina

Accumulating research has indicated that the transplantation of combined stem cells and scaffolds is an effective method for spinal cord injury(SCI).The development of three-dimensional(3D)bioprinting technology can make the 3D scaffolds combined with cells more accurate and effective for SCI ***,unmyelinated newborn nerve fibers have no nerve signaling conduction,hampering recovery of motor *** this study,we designed and printed a type of sodium alginate/gelatin scaffold loaded with neural stem cells and oligodendrocytes,which were involved in the formation of the myelin sheaths of neural cell *** order to observe the effectiveness of this 3D bioprinting scaffold,we transplanted it into the completely transected rat spinal cord,and then immunofluorescence staining,hematoxylin–eosin staining and behavioral assessment were *** results showed that this 3D bioprinting scaffold markedly improved the hindlimb motor function and promoted nerve *** findings suggested that this novel 3D bioprinting scaffold was a good carrier for cells transplantation,thereby enhancing spinal cord repair following injury.

关键词： three-dimensional bioprinting neural stem cell oligodendrocytes spinal cord injury

A coaxially extruded heterogeneous core-shell fiber with Schwann cells and neural stem cells

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Regenerative Biomaterials 2020年第2期7卷 131-139页

作者： Xinda Li Dezhi Zhou Zhizhong Jin Hongqing Chen Xuanzhi Wang Xinzhi Zhang Tao Xu Biomanufacturing and Rapid Forming Technology Key Laboratory of Beijing Department of Mechanical EngineeringTsinghua UniversityBeijing 100084People’s Republic of China Key Laboratory for Advanced Materials Processing Technology Ministry of EducationDepartment of Mechanical EngineeringTsinghua UniversityBeijing 100084People’s Republic of China Department of Neurosurgery the First Hospital of China Medical UniversityShenyang 110122People’s Republic of China Department of Neurosurgery Xijing HospitalFourth Military Medical UniversityXi’an 710032People’s Republic of China Department of Neurosurgery the First Affiliated Hospital of Wannan Medical CollegeWuhu 241001People’s Republic of China East China Institute of Digital Medical Engineering Shangrao 334000People’s Republic of China Medprin Regenerative Medical Technologies Co. LtdShenzhen 518102People’s Republic of China Department of Precision Medicine and Healthcare Tsinghua Berkeley Shenzhen InstituteShenzhen 518055People’s Republic of China

cellular therapies play a critical role in the treatment of spinal cord injury(SCI).Compared with cell-seeded conduits,fully cellular grafts have more similarities with autografts,and thus might result in better regeneration *** this study,we fabricated Schwann cell(SC)-neural stem cell(NSC)core–shell alginate hydrogel fibers in a coaxial extrusion *** rat SC line RSC96 and mouse NSC line NE-4C were used in this *** cellular components were achieved in the core portion and the relative spatial positions of these two cells partially mimic the construction of nerve fibers in *** were demonstrated to express more genes of neurotrophic factors in alginate *** proliferation and differentiation tendency of NSCs was observed when they were co-cultured with *** model has strong potential for application in SCI repair.

关键词： coaxial extrusion neural stem cell Schwann cell

Isolation,Culture and Identification of neural stem cells in New-born Rats

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2004年第1期24卷 75-78页

作者：谢宗平郑启新郭晓东易诚青吴永超 Department of Orthopaedics

The cortexes were obtained from new born rats and dissociated to single cells by triturating. The cells were cultured in neural stem cell (NSC) culture medium (DMEM supplemented with bFGF, EGF and B27) and formed primary neurospheres after 7 days. Single cells dissociated from neurosphere were cultured in 96 well plates and formed single cell cloning neurosphere 7 days later. The primary and single cell cloning neurospheres were both positive for the immunofluorescent staining of nestin and were identified as NSC. It was proved that NSC can be expanded in vitro and provide seed cells for neural tissue engineering.

关键词： neural stem cell cell culture tissue engineering

EXPERIMENTAL STUDY ON PLASTICITY OF PROLIFERATED neural stem cellS IN ADULT RATS AFTER CEREBRAL INFARCTION

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Chinese Medical Sciences Journal 2006年第3期21卷 184-188页

作者： Bo Zhang Ren-zhi Wang Zhi-gang Lian Yang Song Yong Yao Department of Neurosurgery the First Affiliated Hospital of Dalian Medical University Dalian 116011 Department of Neurosurgery Peking Union Medical College Hospital Chinese Academy of Medical Sciences ＆ Peking Union Medical College Beijing 100730

Objective To investigate whether there is endogenous neural stem cell proliferation and whether these proliferated neural stem cells represent neural plasticity in the adult rats after cerebral infarction. Methods Cerebral infarction models of rats were established and the dynamic expression of bromodeoxyuridine （BrdU）, BrdU/polysialylated neural cell adhesion molecule （PSA-NCAM） were determined by immunohistochemistry and immunofluorescence staining. BrdU was used to mark dividing neural stem cells. PSA-NCAM was used to mark the plasticity of neural stem cells. Results Compared with controls, the number of BrdU-positive cells in the subventricular zone （SVZ） and hippocampus increased significantly at 1st day after cerebral infarction （P 〈 0. 05 ）, reached maximum at 7th day, decreased markedly at 14th day, but it was still elevated compared with that of the controls （ P 〈 0. 05 ）. The number of BrdU-labeled with PSA-NCAM-positive cells increased significantly at 7th day （ P 〈 0. 05 ）, reached maximum at 14th day, markedly decreased at 28th day, but it was still elevated compared with that of the controls （P 〈 0. 05 ）. It was equal to 60% of the number of BrdU-positive cells in the same period. Conclusion Cerebral infarction may stimulate the proliferation of endogenous neural stem cells in situ and most proliferated neural stem cells represent neural plasticity.

关键词： cerebral infarction neural stem cell proliferation plasticity

PROLIFERATION AND DIFFERENTIATION OF neural stemcellS IN ADULT RATS AFTER CEREBRAL INFARCTION

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Chinese Medical Sciences Journal 2004年第2期19卷 73-77页

作者： BoZhang Ren-zhiWang YongYao Zhi-haiLiu Zhi-gangLian Yu-jieZou Yu-kuiWei DepartmentofNeurosurgery PekingUnionMedicalCollegeHospitalChineseAcademyofMedicalSciences＆PekingUnionMedicalCollegeBeijing100730 DepartmentofSurgery NanxunPeople＇sHospitalHuzhou313009 DepartmentofNeurosurgery theFirstAffiliatedHospitalofDalianMedicalUniversityDalian116011

Objective To investigate proliferation and differentiation of neural stem cells in adult rats after cerebral infarction. Methods Models of cerebral infarction in rats were made and the time-course expression of bromodeoxyuridine(BrdU), Musashi1, glial fibrillary acidic protein (GFAP), and neuronal nuclear antigen (NeuN) were determined by immunohistochemistry and immunofluorescence staining. BrdU and Musashi1 were used to mark dividing neural stem cells. GFAP and NeuN were used to mark differentiating neural stem cells. Results Compared with controls, the number of BrdU-labeled and BrdU-labeled with Musashi1-positive cells incre-ased strikingly 1 day after cerebral infarction; approximately 6 fold with a peak 7 days later; markedly decreased 14 days later, but was still elevated compared with that of controls; decling to the control level 28 days later. The number of BrdU-labeled with GFAP-positive cells nearly remained unchanged in the hippocampus after cerebral infarction. The nu-mber of BrdU-labeled with NeuN-positive cells increased strikingly 14 days after cerebral infarction, reached maximum peak in the hippocampus 28 days after cerebral infarction in rats. Conclusion Cerebral infarction stimulate proliferation of inherent neural stem cells and most proliferated neural stem cells differentiate into neurons.

关键词： cerebral infarction neural stem cell proliferation differentiation

Identification and culture of neural stem cells isolated from adult rat subventricular zone following fluid percussion brain injury

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Journal of Pharmaceutical Analysis 2010年第3期22卷 175-178页

作者： Ping-Lin Yang1,Jian-Qiang Qu2,Xi-Jing He1,Hao-Peng Li1 1.the Second Department of Orthopaedics 2.Department of Neurosurgery,the Second Affiliated Hospital,Medical School of Xi’an Jiaotong University,Xi’an 710004,China Second Department of Orthopaedics Second Affiliated Hospital Medical School of Xi'an Jiaotong University Xi'an 710004 China Department of Neurosurgery Second Affiliated Hospital Medical School of Xi'an Jiaotong University Xi'an 710004 China

Objective To analyze proliferation and differentiation of glial fibrillary acid protein(GFAP)-and nestin-positive(GFAP+/nestin+)cells isolated from the subventricular zone following fluid percussion brain injury to determine whether GFAP+/nestin+ cells exhibit characteristics of neural stem *** Male Sprague-Dawley rats,aged 12 weeks and weighing 200-250 g,were randomly and evenly assigned to normal control group and model *** the model group,a rat model of fluid percussion brain injury was *** days later,subventricular zone tissue was resected from each group and made into single cell *** serum-free neural stem cell medium culture and subsequent serum-induced differentiation,cell type,proliferation and differentiation capacities were determined by immunofluorescence staining and flow *** At 3-7 days after fluid percussion brain injury,nestin+/GFAP+ cells in the single cell suspension from the model group significantly outnumbered those from the normal control group(Pcell edge and bulged center formed after primary culture,and were clearly visible with the increase of culture time and medium *** several passages,many clonal spheres were obtained,suggesting strong self-proliferatiing *** from the model group differentiated into astrocytes,neurons and *** GFAP+/nestin+ cells isolated from the adult rat subventricular zone after fluid percussion brain injury are thought to be neural stem cells because of their self-renewal and multi-differentiation capacities.

关键词： brain injury astrocyte neural stem cell subventricular zone

Gene expression profiling and mechanism study of neural stem cells response to surface chemistry

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Regenerative Biomaterials 2014年第1期1卷 37-47页

作者： Ying Wang Shenglian Yao Qingyuan Meng Xiaolong Yu Xiumei Wang Fuzhai Cui Institute for regenerative medicine and biomimetic materials School of materials science and engineeringTsinghua UniversityBeijing 100084China Department of anatomy histology and embryologySchool of basic medical sciencesCapital Medical UniversityBeijing 100069China Department of material science and chemical engineering Hainan UniversityHaikou 570228China

To declare the mechanisms of neural stem cells(NSCs)in response to material surface chemistry,NSCs were exposed to the self-assemble monolayers of alkanethiolates on gold surfaces terminated with amine(NH2),hydroxyl(OH)and methyl(CH3)for *** morphological responses of NSCs were recorded;the gene expression profilings were detected by genechips;the gene expressions data of NSCs responded to different chemical groups were declared through the gene ontology term and pathway *** showed that cells behaved dissimilar on the three chemical groups,the adhesion,proliferation and migration were easier on the NH2 and OH groups;the gene expressions of NSCs were induced differently,either,involved in several functional processes and signaling ***3 group induced genes enriched much in chemistry reactions and death processes,whereas many genes of cellular nucleotide metabolism were ***2 group induced NSCs to express many genes of receptors on membrane,and participated in cellular signal transduction of cell adhesion and interactions,or associated with axon *** group was similar to NH2 group to induce the membrane response,but it also down regulated metabolism of ***,it declared the chemical groups affected NSCs through inner way and the NH2,OH and CH3 groups triggered the cellular gene expression in different signaling pathways.

关键词： neural stem cell chemical group biomaterial gene expression receptor signaling pathway

Synergistic Effect of Schwann cells and Retinoic Acid on Differentiation and Synaptogenesis of Hippocampal neural stem cells in vitro

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Biomedical and Environmental Sciences 2006年第3期19卷 219-224页

作者： XUE-BAO ZHANG YUAN-SHAN ZENG WEI ZHANG YA-YUN CHEN YI XIONG SUI-JUN CHEN Division of Neuroscience Department of Histology and Embryology Zhongshan Medical College Sun Yat-Sen University Guangzhou 510080 Guangdong China

Objective To investigate the synergistic effect of Schwann cells （SCs） and retinoic acid （R.A） on differentiation and synaptogenesis of neural stem cells （NSCs） derived from hippocampus of neonatal rats. Methods The classical method for 2×2 factorial analysis experiment was used to assess synergistic action of SCs and RA. NSCs were treated with R.A, SCs, and SCs ＋ RA in DMEM/F12 with 0.5% fetal bovine serum for six days, respectively. Double immunofluorescent staining was used to detect the differentiation of NSCs including nestin, glial fibrillary acidic protein （GFAP） and Map2. The expression of PSD95 was used to demonstrate synaptogenesis. Results After NSCs were treated with RA or SCs, the expression of nestin and GFAP was significantly decreased while the expression of Map2 and PSD95 was significantly increased in comparison with the control. Factorial ANOVA showed that interactions between SCs and RA could induce the expression of Map2 and PSD95. Conclusion SCs and RA could promote synergistically the neuronal differentiation and synaptogenesis of hippocampal neural stem cells in vitro while they decreased the astrocytes and nestin positive NSCs.

关键词： neural stem cell Neuronal differentiation PSD95 Schwann cells Retinoic acid

Ultrasound microbubbles combined with liposome-mediated pNogo-R shRNA delivery into neural stem cells

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neural Regeneration Research 2012年第1期7卷 54-59页

作者： Weixia Ye Xueping Huang Yangyang Sun Hao Liu Jin Jiang Youde Cao Department of Pathology Molecular Medicine and Cancer Research Center Chongqing Medical University Chongqing 400016 China Department of Orthopaedics Luzhou People＇s Hospital Luzhou 646000 Sichuan Province China

In the present study, ultrasound-mediated microbubble destruction （UMMD） alone and combined with liposome technology was used as a novel nonviral technique to transfect a Nogo receptor （Nogo-R） shRNA plasmid （pNogo-R shRNA） into neural stem cells （NSCs）. Using green fluorescent protein as a reporter gene, transfection efficiency of NSCs was significantly higher in the group transfected with UMMD combined with liposomes compared with that of the group transfected with UMMD or liposomes alone, and did not affect cell vitality. In addition, Nogo-R mRNA and protein expression was dramatically decreased in the UMMD combined with liposome-mediated group compared with that of other groups after 24 hours of transfection. The UMMD technique combined with liposomes is a noninvasive gene transfer method, which showed minimal effects on cell viability and effectively increased transfer of Nogo-R shRNA into NSCs.

关键词： ultrasound microbubble liposome neural stem cell gene transfection Nogo receptor neural regeneration