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The combined application of stem cells and three-dimensional bioprinting scaffolds for the repair of spinal cord injury

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Neural Regeneration Research 2024年第8期19卷 1751-1758页

作者： Dingyue Ju Chuanming Dong Department of Anatomy Medical College of Nantong UniversityNantongJiangsu ProvinceChina key laboratory of neuroregeneration of Jiangsu and Ministry of Education Co-innovation Center of NeuroregenerationNantong UniversityNantongJiangsu ProvinceChina

Spinal cord injury is considered one of the most difficult injuries to repair and has one of the worst prognoses for injuries to the nervous system.Following surgery,the poor regenerative capacity of nerve cells and the generation of new scars can make it very difficult for the impaired nervous system to restore its neural functionality.Traditional treatments can only alleviate secondary injuries but cannot fundamentally repair the spinal cord.Consequently,there is a critical need to develop new treatments to promote functional repair after spinal cord injury.Over recent years,there have been seve ral developments in the use of stem cell therapy for the treatment of spinal cord injury.Alongside significant developments in the field of tissue engineering,three-dimensional bioprinting technology has become a hot research topic due to its ability to accurately print complex structures.This led to the loading of three-dimensional bioprinting scaffolds which provided precise cell localization.These three-dimensional bioprinting scaffolds co uld repair damaged neural circuits and had the potential to repair the damaged spinal cord.In this review,we discuss the mechanisms underlying simple stem cell therapy,the application of different types of stem cells for the treatment of spinal cord injury,and the different manufa cturing methods for three-dimensional bioprinting scaffolds.In particular,we focus on the development of three-dimensional bioprinting scaffolds for the treatment of spinal cord injury.

关键词： biomaterials embryonic stem cells induced pluripotent stem cells mesenchymal stem cells nerve regeneration spinal cord injury stem cell therapy stem cells three-dimensional bioprinting

Chemokine platelet factor 4 accelerates peripheral nerve regeneration by regulating Schwann cell activation and axon elongation

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Neural Regeneration Research 2024年第1期19卷 190-195页

作者： Miao Gu Xiao Cheng Di Zhang Weiyan Wu Yi Cao Jianghong He key laboratory of neuroregeneration of Jiangsu and Ministry of Education Co-innovation Center of NeuroregenerationNMPA Key Laboratory for Research and Evaluation of Tissue Engineering Technology ProductsNantong UniversityNantongJiangsu ProvinceChina School of Basic Medical Sciences Hebei Key Laboratory of Nerve Injury and RepairChengde Medical UniversityChengdeHebei ProvinceChina

Schwann cells in peripheral nerves react to traumatic nerve injury by attempting to grow and regenerate.Howeve r,it is unclear what factors play a role in this process.In this study,we searched a GEO database and found that expression of platelet factor 4 was markedly up-regulated after sciatic nerve injury.Platelet factor is an important molecule in cell apoptosis,diffe rentiation,survival,and proliferation.Further,polymerase chain reaction and immunohistochemical staining confirmed the change in platelet factor 4 in the sciatic nerve at different time points after injury.Enzyme-linked immunosorbent assay confirmed that platelet factor 4 was secreted by Schwann cells.We also found that silencing platelet factor 4 decreased the proliferation and migration of primary cultured Schwann cells,while exogenously applied platelet factor 4 stimulated Schwann cell prolife ration and migration and neuronal axon growth.Furthermore,knocking out platelet factor 4 inhibited the prolife ration of Schwann cells in injured rat sciatic nerve.These findings suggest that Schwann cell-secreted platelet factor 4 may facilitate peripheral nerve repair and regeneration by regulating Schwann cell activation and axon growth.Thus,platelet factor 4 may be a potential therapeutic target for traumatic peripheral nerve injury.

关键词： axon elongation bioinformatic analysis cell migration cell proliferation dorsal root ganglia peripheral nerve regeneration peripheral nerve trauma platelet factor 4 rat sciatic nerve Schwann cells

Biocompatibility studies of silk fibroin-based artificial nerve grafts in vitro and in vivo

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Progress in Natural Science:Materials International 2007年第9期17卷 1029-1034页

作者： Chen Xuemei,Yang Yumin,Wu Jian,Zhao Yahong,Ding Fei and Gu Xiaosong(Jiangsu key laboratory of neuroregeneration,Nantong University,Nantong 226001,China) Jiangsu key laboratory of neuroregeneration Nantong University Nantong 226001 China

Silk fibroin(SF)has been used extensively in the biomedical field including tissue engineering for the generation of artificial bones,skins or ligaments.We have previously reported on good in vitro biocompatibility of SF fibers with peripheral nerve tissues and cells.In the present study,we developed a novel design of the SF-based artificial nerve graft(SF graft)which was composed of a SF-nerve guidance conduit(NGC)inserted with SF fibers.MTT assay was performed to determine the cytotoxicity of the SF-NGC extract fluid on the cultured L929 cells derived from an immortalized mouse fibroblast cell line.In addition,this SF graft was implanted into adult rats for bridging a 10-mm long sciatic nerve defect.The following-up experiments at initial stage(1-4 week)of nerve regeneration including routine blood tests and histochemical investigation were conducted to evaluate the in vivo biocompatibility of the SF graft with peripheral nerves.The results demonstrated that the SF-NGC graft was biocompatible with the surrounding tissues and cells due to its low inflammatory potential with a grade 0 under the U.S.Pharmacopeia guidelines and it was generally suitable to a certain degree for bridging peripheral nerve defects in virtue of supporting Schwann cell adherence,expansion and migration.Therefore the SF graft is a promising alternative to classical autografts for peripheral nerve repair.

关键词： silk fibroin artificial nerve graft nerve guidance conduit biocompatibility cytotoxicity

Nerve regeneration factor promotes nerve regeneration in rat

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Progress in Natural Science:Materials International 2006年第12期16卷 1281-1287页

作者： ZHANG Qi, LIU Mei, ZHOU Mingming and DING Fei**(key laboratory of neuroregeneration, Nantong University, Nantong 226001, China ) key laboratory of neuroregeneration Nantong University Nantong 226001 China

Nerve regeneration factor (NRF) extracted from an oral liquid of traditional Chinese medicine, as a nerve growth decoction has been reported by previous studies to exert effects of promoting nerve growth and preventing neuron apoptosis. For new insights into the function of NRF on primary cultured neurons, we investigated the neurite outgrowth in cultured rat dorsal root ganglion (DRG) explant treated with NRF by immunofluorescence and the gene and protein expressions of neurofilament-H(NF-H) and growth associated protein 43 (GAP43) in the cultured rat DRG neurons by real-time quantitative RT-PCR and Western blotting, respectively. In addition, we used a rat model of sciatic nerve crush to evaluate the effect of NRF on regeneration of injured sciatic nerve by a combination of walk track analysis, electrophysiological and histological assessments. The in vitro experiments indicated that NRF promoted the neurite growth of DRGs and the expression of NF-H and GAP43 at mRNA and protein levels in DRG neurons, and in vivo experiments showed that NRF improved peripheral nerve regeneration and functional recovery.

关键词： nerve regeneration factor dorsal root ganglion sciatic nerve crush injury neurofilament-H growth associated protein 43.

Achyranthes bidentata Blume extract promotes neuronal growth in cultured embryonic rat hippocampal neurons

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Progress in Natural Science:Materials International 2009年第5期19卷 549-555页

作者： Xin Tang1, Yiren Chen 1, Xiaosong Gu, Fei Ding Jiangsu key laboratory of neuroregeneration, Nantong University, Nantong 226001, China Jiangsu key laboratory of neuroregeneration Nantong University Nantong 226001 China

We have prepared an aqueous extract of Achyranthes bidentata Blume, a commonly prescribed Chinese medicinal herb, and reported, in previous studies, that A. bidentata extract benefits nerve growth and prevents neuron apoptosis. In this study, we investigated the actions of A. bidentata extract on survival and growth of primarily cultured rat hippocampal neurons. The morphological observation revealed that neurite growth from hippocampal neurons was significantly enhanced by A. bidentata extract with similar effects to those induced by nerve growth factor (NGF), and the greatest neurite growth appeared on treatment with A. bidentata extract at 1 lg/ml for 24 h. DNA microarray analysis indicated that there were 25 upregulated genes and 47 downregulated genes exhibiting significantly differential expression in hippocampal neurons treated with A. bidentata extract at 1 lg/ml for 6 h when compared to those in untreated hippocampal neurons. Real-time quantitative RT-PCR and Western blot analysis demonstrated that the expression of growth-associated protein-43 in hippocampal neurons was upregulated at both mRNA and protein levels after treatment with A. bidentata extract, and the optimal dosage of the extract was also 1 lg/ml. These data confirm that A. bidentata extract could promote in vitro hippocampal neuronal growth in a dose-and time-dependent manner.

关键词： Achyranthes bidentata Blume Aqueous extract Nerve growth factor Hippocampal neurons Neurite outgrowth Growth-associated protein-43 Gene differential expression

Tau truncation in the pathogenesis of Alzheimer's disease: a narrative review

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Neural Regeneration Research 2024年第6期19卷 1221-1232页

作者： Dandan Chu Xingyue Yang Jing Wang Yan Zhou Jin-Hua Gu Jin Miao Feng Wu Fei Liu key laboratory of neuroregeneration of Jiangsu and Ministry of Education NMPA Key Laboratory for Research and Evaluation of Tissue Engineering Technology Products Coinnovation Center of Neuroregeneration Nantong University 2. Department of Pharmacology School of Pharmacy Nantong University 3. Department of Biochemistry and Molecular Biology School of Medicine Nantong University 4. Department of Clinical Pharmacy Affiliated Maternity and Child Health Care Hospital of Nantong University Nantong University 5. Laboratory of Animal Center Nantong University Department of Neurochemistry Inge Grundke-Iqbal Research Floor New York State Institute for Basic Research in Developmental Disabilities

Alzheimer’s disease is characterized by two major neuropathological hallmarks—the extracellular β-amyloid plaques and intracellular neurofibrillary tangles consisting of aggregated and hyperphosphorylated Tau protein. Recent studies suggest that dysregulation of the microtubuleassociated protein Tau, especially specific proteolysis, could be a driving force for Alzheimer’s disease neurodegeneration. Tau physiologically promotes the assembly and stabilization of microtubules, whereas specific truncated fragments are sufficient to induce abnormal hyperphosphorylation and aggregate into toxic oligomers, resulting in them gaining prion-like characteristics. In addition, Tau truncations cause extensive impairments to neural and glial cell functions and animal cognition and behavior in a fragment-dependent manner. This review summarizes over 60 proteolytic cleavage sites and their corresponding truncated fragments, investigates the role of specific truncations in physiological and pathological states of Alzheimer’s disease, and summarizes the latest applications of strategies targeting Tau fragments in the diagnosis and treatment of Alzheimer’s disease.

关键词： Alzheimer’s disease cleavage site diagnosis marker neurofibrillary tangles phosphorylation Tau Tau aggregation therapy truncation

Role of CD36 in central nervous system diseases

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Neural Regeneration Research 2024年第3期19卷 512-518页

作者： Min Feng Qiang Zhou Huimin Xie Chang Liu Mengru Zheng Shuyu Zhang Songlin Zhou Jian Zhao School of Health Science and Engineering University of Shanghai for Science and TechnologyShanghaiChina Department of Orthopedic Oncology Second Affiliated Hospital of Naval Medical UniversityShanghaiChina key laboratory of neuroregeneration of Jiangsu and Ministry of Education NMPA Key Laboratory for Research and Evaluation of Tissue Engineering Technology ProductsJiangsu Clinical Medicine Center of Tissue Engineering and Nerve Injury RepairCo-Innovation Center of NeuroregenerationNantong UniversityNantongJiangsu ProvinceChina Department of Stomatology Affiliated Hospital of Nantong UniversityNantongJiangsu ProvinceChina Medical College of Nantong University NantongJiangsu ProvinceChina

CD36 is a highly glycosylated integral membrane protein that belongs to the scavenger receptor class B family and regulates the pathological progress of metabolic diseases.CD36 was recently found to be widely expressed in various cell types in the nervous system,including endothelial cells,pericytes,astrocytes,and microglia.CD36 mediates a number of regulatory processes,such as endothelial dysfunction,oxidative stress,mitochondrial dysfunction,and inflammatory responses,which are involved in many central nervous system diseases,such as stroke,Alzheimer’s disease,Parkinson’s disease,and spinal cord injury.CD36 antagonists can suppress CD36 expression or prevent CD36 binding to its ligand,thereby achieving inhibition of CD36-mediated pathways or functions.Here,we reviewed the mechanisms of action of CD36 antagonists,such as Salvianolic acid B,tanshinone IIA,curcumin,sulfosuccinimidyl oleate,antioxidants,and small-molecule compounds.Moreover,we predicted the structures of binding sites between CD36 and antagonists.These sites can provide targets for more efficient and safer CD36 antagonists for the treatment of central nervous system diseases.

关键词： animal experiments antagonists CD36 antagonist central nervous system diseases clinical trial curcumin microRNA salvianolic acid B small-molecule drugs sulfosuccinimidyl oleate

RNA sequencing of exosomes secreted by fibroblast and Schwann cells elucidates mechanisms underlying peripheral nerve regeneration

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Neural Regeneration Research 2024年第8期19卷 1812-1821页

作者： Xinyang Zhou Yehua Lv Huimin Xie Yan Li Chang Liu Mengru Zheng Ronghua Wu Songlin Zhou Xiaosong Gu Jingjing Li Daguo Mi Suzhou Medical College of Soochow University SuzhouJiangsu ProvinceChina key laboratory of neuroregeneration of Jiangsu and Ministry of Education Co-Innovation Center of NeuroregenerationNantong UniversityNantongJiangsu ProvinceChina Department of Orthopedic Nantong Traditional Chinese Medicine HospitalNantongJiangsu ProvinceChina Nantong Stomatological Hospital Affiliated to Nantong University NantongJiangsu ProvinceChina Department of General Practice Affiliated Hospital of Nantong UniversityNantongJiangsu ProvinceChina

Exosomes exhibit complex biological functions and mediate a variety of biological processes,such as promoting axonal regeneration and functional recove ry after injury.Long non-coding RNAs(IncRNAs)have been reported to play a crucial role in axonal regeneration.Howeve r,the role of the IncRNA-microRNAmessenger RNA(mRNA)-competitive endogenous RNA(ceRNA)network in exosome-mediated axonal regeneration remains unclear.In this study,we performed RNA transcriptome sequencing analysis to assess mRNA expression patterns in exosomes produced by cultured fibroblasts(FC-EXOs)and Schwann cells(SCEXOs).Diffe rential gene expression analysis,Gene Ontology analysis,Kyoto Encyclopedia of Genes and Genomes analysis,and protein-protein intera ction network analysis were used to explo re the functions and related pathways of RNAs isolated from FC-EXOs and SC-EXOs.We found that the ribosome-related central gene Rps5 was enriched in FC-EXOs and SC-EXOs,which suggests that it may promote axonal regeneration.In addition,using the miRWalk and Starbase prediction databases,we constructed a regulatory network of ceRNAs targeting Rps5,including 27 microRNAs and five IncRNAs.The ceRNA regulatory network,which included Ftx and Miat,revealed that exsosome-derived Rps5 inhibits scar formation and promotes axonal regeneration and functional recovery after nerve injury.Our findings suggest that exosomes derived from fibro blast and Schwann cells could be used to treat injuries of peripheral nervous system.

关键词： ceRNA network exosomes fibroblast cells Gene Ontology(GO) Kyoto Encyclopedia of Genes and Genomes(KEGG) protein-protein interaction(PPI)networks RNA-seq Schwann cells

Protein expression profile in the differentiation of rat bone marrow stromal cells into Schwann cell-like cells

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Science China(Life Sciences) 2009年第3期52卷 267-277页

作者： LI WenTing1,SUN HuaLin2,XU ZengLu1,DING Fei2 & GU XiaoSong2 1 Department of Anatomy,Histology and Embryology,Peking Union Medical College,Institute of Basic Medical Science,Chinese Academy of Medical Sciences,Beijing 100005,China 2 Jiangsu key laboratory of neuroregeneration,Nantong University,Nantong 226001,China Department of Anatomy Histology and Embryology Peking Union Medical College Institute of Basic Medical Science Chinese Academy of Medical Sciences Beijing China Jiangsu key laboratory of neuroregeneration Nantong University Nantong China

During the last decade,increasing evidence suggested that bone marrow stromal cells(MSCs) have the potential to differentiate into neural lineages.Many studies have reported that MSCs showed morphological changes and expressed a limited number of neural proteins under experimental conditions.However,no proteomic studies on MSCs differentiated into Schwann cell-like cells have been reported.In this study,we isolated MSCs from adult Sprague-Dawley rat femur and tibia bone marrows and induced the cells in vitro under specific conditions.By using two-dimensional gel electrophoresis(2-DE),we compared the protein profiles of MSCs before and after induced differentiation.We obtained 792 protein spots in the protein profile by 2-DE,and found that 74 spots changed significantly before and after the differentiation using PDQuest software,with 43 up-regulated and 31 down-regulated.We analyzed these 74 spots by a matrix assisted laser desorption ionization-time of flight mass spectrometry(MALDI-TOF-MS) and by database searching,and found that they could be grouped into various classes,including cytoskeleton and structure proteins,growth factors,metabolic proteins,chaperone proteins,receptor proteins,cell cycle proteins,calcium binding proteins,and other proteins.These proteins also include neural and glial proteins,such as BDNF,CNTF and GFAP.The results may provide valuable proteomic information about the differentiation of MSCs into Schwann cell-like cells.

关键词： one marrow stromal cells(MSCs) differentiation Schwann cell-like cells proteomics