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Cardiac patches made of brown adipose-derived stem cell sheets and conductive electrospun nanofibers restore infarcted heart for ischemic myocardial infarction

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Bioactive Materials 2023年第9期27卷 271-287页

作者： Xinbo Wei Li Wang cuimi duan Kai Chen Xia Li Ximin Guo Peng Chen Haifeng Liu Yubo Fan Key Laboratory for Biomechanics and Mechanobiology(Beihang University)of Ministry of Education Beijing Advanced Innovation Center for Biomedical EngineeringSchool of Biological Science and Medical EngineeringBeihang UniversityBeijing100083PR China Department of Advanced Interdisciplinary Studies Institute of Basic Medical SciencesBeijing100850PR China Beijing Citident Stomatology Hospital Beijing100032PR China Department of Ultrasound The Third Medical CenterChinese PLA General HospitalBeijingPR China

Cell sheet engineering has been proven to be a promising strategy for cardiac remodeling post-myocardial infarction. However, insufficient mechanical strength and low cell retention lead to limited therapeutic efficiency. The thickness and area of artificial cardiac patches also affect their therapeutic efficiency. Cardiac patches prepared by combining cell sheets with electrospun nanofibers, which can be transplanted and sutured to the surface of the infarcted heart, promise to solve this problem. Here, we fabricated a novel cardiac patch by stacking brown adipose-derived stem cells (BADSCs) sheet layer by layer, and then they were combined with multi-walled carbon nanotubes (CNTs)-containing electrospun polycaprolactone/silk fibroin nanofibers (CPSN). The results demonstrated that BADSCs tended to generate myocardium-like structures seeded on CPSN. Compared with BADSCs suspension-containing electrospun nanofibers, the transplantation of the CPSN-BADSCs sheets (CNBS) cardiac patches exhibited accelerated angiogenesis and decreased inflammation in a rat myocardial infarction model. In addition, the CNBS cardiac patches could regulate macrophage polarization and promote gap junction remodeling, thus restoring cardiac functions. Overall, the hybrid cardiac patches made of electrospun nanofibers and cell sheets provide a novel solution to cardiac remodeling after ischemic myocardial infarction.

关键词： Myocardial infarction brown adipose-derived stem cells Cell sheet engineering Electrospun nanofibers Polycaprolactone/silk fibroin Carbon nanotubes

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Embryoid bodies formation and differentiation from mouse embryonic stem cells in collagen/Matrigel scaffolds

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Journal of Genetics and Genomics 2010年第7期37卷 451-460页

作者： Jin Zhou Ye Zhang Qiuxia Lin Zhiqiang Liu Haibin Wang cuimi duan Yanmeng Wang Tong Hao Kmwu Wu Changyong Wang Department of Tissue Engineering Institute of Basic Medical Sciences and Tissue Engineering Research Center Academy of Military Medical Sciences Beijing 100850 China Department of Hematology and Oncology General Hospital of the Artillery Force Second Beijing 100850 China

Embryonic stem （ES） cells have the potential to develop into any type of tissue and are considered as a promising source of seeding cells for tissue engineering and transplantation therapy.The main catalyst for ES cells differentiation is the growth into embryoid bodies （EBs）,which are utilized widely as the trigger of in vitro differentiation.In this study,a novel method for generating EBs from mouse ES cells through culture in collagen/Matrigel scaffolds was successfully established.When single ES cells were seeded in three dimensional collagen/Matrigel scaffolds,they grew into aggregates gradually and formed simple EBs with circular structures.After 7 days＇ culture,they formed into cystic EBs that would eventually differentiate into the three embryonic germ layers.Evaluation of the EBs in terms of morphology and potential to differentiate indicated that they were typical in structure and could generate various cell types;they were also able to form into tissue-like structures.Moreover,with introduction of ascorbic acid,ES cells differentiated into cardiomyocytes efficiently and started contracting synchronously at day 19.The results demonstrated that collagen/Matrigel scaffolds supported EBs formation and their subsequent differentiation in a single three dimensional environment.

关键词： embryonic stem （ES） cells embryoid bodies （EBs） differentiation collagen/Matrigel scaffolds model

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Identification and characterization of mammaglobin-A epitope in heterogenous breast cancers for enhancing tumor-targeting therapy

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Signal Transduction and Targeted Therapy 2020年第1期5卷 1778-1788页

作者： Zhiqiang Liu Xiqin Yang cuimi duan Jiangxue Li Rongsheng Tong Yuting Fan Jiannan Feng Ruiyuan Cao Wu Zhong Xiaoyan Feng Heqiu Zhang Lulu Cai Beijing Institute of Basic Medical Sciences 27 Taiping RoadHaidian DistrictBeijing 100850China Personalized Drug Therapy Key Laboratory of Sichuan Province Department of PharmacySichuan Provincial People’s HospitalUniversity of Electronic Science and Technology of ChinaChengdu 611731China Beijing Institute of Pharmacology and Toxicology 27 Taiping RoadHaidian DistrictBeijing 100850China

Although targeted therapy has been extensively investigated for breast cancers,a molecular target with broad application is currently unavailable due to the high heterogeneity of these cancers.Mammaglobin-A(Mam-A),which is overexpressed in most breast carcinomas,has been proposed as a promising target.However,the lack of specific targeting moieties due to uncertain binding epitopes hampers further translational study.Here,seven potential epitopes of Mam-A were disclosed,and a unique epitope was then identified in most types of breast cancers,despite the genotypic heterogeneity.With phage display technology,the epitope was determined to be N-terminal amino acids 42–51 of Mam-A(N42–51).Then,the N42–51 epitope-specific monoclonal antibody,mAb785,was conjugated to poly lactic-co-glycolic acid(PLGA)nanoparticles loaded with therapeutic agents,thereby enhancing the drug uptake and therapeutic efficacy in different genotypes of breast cancers.The computer simulation of the N42–51 epitope and the mAb785 structures,as well as their interactions,further revealed the specific targeting mechanism of the mAb785-conjugated nanoparticles to breast cancers.

关键词： breast cancers epitope

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Distribution and characteristics of telocytes as nurse cells in the architectural organization of engineered heart tissues

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Science China(Life Sciences) 2014年第2期57卷 241-247页

作者： ZHOU Jin WANG Yan ZHU Ping SUN HongYu MOU YongChao duan cuimi YAO AnNing LV ShuangHong WANG ChangYong Department of Advanced Interdisciplinary Studies Institute of Basic Medical Sciences and Tissue Engineering Research CenterAcademy of Military Medical Sciences The First Department of Cardiology South Building The General Hospital of People's Liberation Army Laboratory of Oncology Affiliated Hospital of Academy of Military Medical Sciences

Interstitial Cajal-like cells are a distinct type of interstitial cell with a wide distribution in mammalian organs and tissues,and have been given the name"telocytes".Recent studies have demonstrated the potential roles of telocytes in heart development,renewal,and repair.However,further research on the functions of telocytes is limited by the complicated in vivo environment.This study was designed to construct engineered heart tissue(EHT)as a three-dimensional model in vitro to better understand the role of telocytes in the architectural organization of the myocardium.EHTs were constructed by seeding neonatal cardiomyocytes in collagen/Matrigel scaffolds followed by culture under persistent static stretch.Telocytes in EHTs were identified by histology,toluidine blue staining,immunofluorescence,and transmission electron microscopy.The results from histology and toluidine blue staining demonstrated widespread putative telocytes with compact toluidine blue-stained nuclei,which were located around cardiomyocytes.Prolongations from the cell bodies showed a characteristic dichotomous branching pattern and formed networks in EHTs.Immunofluorescence revealed positive staining of telocytes for CD34 and vimentin with typical moniliform prolongations.A series of electron microscopy images further showed that typical telocytes embraced the cardiomyocytes with their long prolongations and exhibited a marked appearance of nursing cardiomyocytes during the construction of EHTs.This finding highlights the great importance of telocytes in the architectural organization of EHTs.It also suggests that EHT is an appropriate physical and pathological model system in vitro to study the roles of telocytes during heart development and regeneration.

关键词：滋养细胞组织架构心脏组织工程化透射电子显微镜心肌细胞特性甲苯胺蓝

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以β-磷酸三钙多孔陶瓷为载体建造组织工程化人工软骨

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中华整形外科杂志 2002年第1期18卷 9-11页

作者：王常勇薄斌赵强郭希民段翠密王永红范明卢建熙军事医学科学院基础医学研究所北京100850 北京军医学院法国地中海大学

目的　通过将软骨细胞接种到三维多孔 β 磷酸三钙 (β TCP)陶瓷支架材料上 ,探讨以β TCP为载体建造组织工程化软骨的可行性。方法　将 β TCP多孔陶瓷加工成圆片状 ,并将其作为构建人工软骨的细胞支架。在支架材料上接种从兔关节软... 详细信息

目的　通过将软骨细胞接种到三维多孔 β 磷酸三钙 (β TCP)陶瓷支架材料上 ,探讨以β TCP为载体建造组织工程化软骨的可行性。方法　将 β TCP多孔陶瓷加工成圆片状 ,并将其作为构建人工软骨的细胞支架。在支架材料上接种从兔关节软骨分离的软骨细胞 ,将细胞———陶瓷复合体置于旋转细胞培养系统 (RCCS)内 ,培养 1周后 ,将其移植到裸鼠背部皮下。植入术后 4 ,8,16周取材 ,进行大体观察、组织学及组织化学等观察。结果　复合体体内移植后 ,在裸鼠皮下有新生软骨形成 ,形成的软骨基本保持了支架材料原有形态。结论　β TCP多孔陶瓷是软骨细胞较适宜的贴附基质 ,以其为支架能够在体内建造出具有精确解剖形状的人工软骨。

关键词：组织工程生物陶瓷软骨细胞人工软骨 β-磷酸三钙陶瓷

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抗坏血酸体外诱导小鼠胚胎干细胞向心肌细胞分化

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解剖学杂志 2007年第4期30卷 392-396页

作者：何文俊王晶波王常勇段翠密李晶吕双红军事医学科学院基础医学研究所组织工程研究室北华大学医学院吉林132013

目的:探讨抗坏血酸作为诱导剂对小鼠胚胎干细胞(mESC)分化为心肌细胞的影响,建立一种体外诱导mESC分化为心肌细胞的实验方法。方法:采用直接悬浮培养法使mESC形成拟胚体(EBs),用含不同浓度抗坏血酸的分化培养基对其进行诱导分化,摸索最... 详细信息

目的:探讨抗坏血酸作为诱导剂对小鼠胚胎干细胞(mESC)分化为心肌细胞的影响,建立一种体外诱导mESC分化为心肌细胞的实验方法。方法:采用直接悬浮培养法使mESC形成拟胚体(EBs),用含不同浓度抗坏血酸的分化培养基对其进行诱导分化,摸索最佳的诱导分化条件。结果:抗坏血酸诱导mESC分化为心肌细胞的最佳浓度为0.1mg/ml,约为70%的EBs出现跳动的心肌合胞体,显著高于不添加任何诱导剂的对照组。抗坏血酸诱导产生的心肌细胞表达多种心肌蛋白,具有心肌细胞的结构特征。结论:抗坏血酸能够促进mESC向心肌细胞分化,应用抗坏血酸体外诱导mESC向心肌细胞分化是一种较为理想的体外诱导方法。

关键词：抗坏血酸胚胎干细胞心肌细胞细胞分化

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兔子宫内膜中标记滞留细胞的初步鉴定

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解剖学杂志 2008年第5期31卷 608-610,F0002页

作者：王海滨段翠密王常勇章烨李德雪郝彤王滟濛吕双红军事医学科学院基础医学研究所组织工程研究室北京100850 第二炮兵总医院肿瘤科北京100088

目的:利用鉴定标记滞留细胞(LRC)的方法分析兔子宫内膜中可能存在的成体干细胞及其分布。方法:首先用促性腺激素处理兔,将子宫内膜调整到增殖期,并采用免疫组织化学方法鉴定增殖期子宫内膜中雌激素受体、孕激素受体、增殖细胞核抗原(PC... 详细信息

目的:利用鉴定标记滞留细胞(LRC)的方法分析兔子宫内膜中可能存在的成体干细胞及其分布。方法:首先用促性腺激素处理兔,将子宫内膜调整到增殖期,并采用免疫组织化学方法鉴定增殖期子宫内膜中雌激素受体、孕激素受体、增殖细胞核抗原(PCNA)的表达,以评价子宫内膜的增殖期状态。BrdU标记后追踪8周,以鉴定LRC细胞的存在与分布。结果:增殖期子宫内膜较促性腺激素处理前明显增厚,子宫内膜中雌激素受体、孕激素受体与PCNA的表达明显升高。8周追踪后,子宫内膜中有(2.1±2.4)%的细胞核为BrdU阳性,即LRC细胞。这些LRC细胞主要存在于腔上皮组织中。结论:在成年兔子宫内膜中有LRC存在,子宫内膜成体干细胞可能存在于这些LRC中。

关键词：促性腺激素子宫内膜标记滞留细胞成体干细胞兔

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Bioceramics-based cartilage tissue engineering

Bioceramics-based cartilage tissue engineering

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The 6th International Conference on Biomedical Engineering and Rehabilitation Engineering

作者： Wang Changyong Quo Ximin Bo Bin Lu Jianxi Zhao Qiang duan cuimi Tissue Engineering Research Center of Academy of Military Medical Sciences Beijing100850 China E-mail: wcy2000@***

＜正＞ Various biomaterials have been developed yet proved to be far from ideal for cartilagetissue engineering. In this study we introduced a new scaffold, bioceramics, to generatecartilage replacement. Beta-TCP porous ceramics seeded with sheep chondrocytes wereimplanted subcutaneously in a nude mice model and in sheep articular cartilage defects.

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人脐带间充质干细胞来源的外泌体促进小鼠成骨前体细胞增殖与分化

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武警医学 2020年第4期31卷 335-339页

作者：王丽段翠密原芳芳王燕郭希民郭红延解放军总医院第三医学中心口腔科北京100039 锦州医科大学研究生院 121000 军事科学院军事医学研究院北京100850

目的提取人脐带间充质干细胞(human umbilical cord mesenchymal stem cells,hucMSCs)来源的外泌体(hucMSC-exo),观察其对小鼠成骨前体细胞(mouse osteoblast progenitor cells,mOPCs)的作用。方法体外分离培养hucMSCs,超速离心法收集外泌体;采用CCK8试剂盒检测不同浓度梯度hucMSC-exo对mOPCs增殖的影响,碱性磷酸酶活性检测以及茜素红染色观察不同浓度梯度hucMSC-exo对mOPCs矿化的影响。结果成功提取hucMSCs,呈现旋涡状贴壁式生长,形态呈长梭形,具有多向诱导分化潜能。HucMSCs来源的外泌体呈圆形、椭圆形,大小在80 nm左右最多,阳性表达CD9,CD63。CCK8法结果显示5、10μg/ml的hucMSC-exo可以显著地促进mOPCs在体外的增殖(P<0.05),碱性磷酸酶和茜素红染色结果发现5、10μg/ml的hucMSC-exo组可以明显提高mOPCs碱性磷酸酶活性及钙化结节的形成,且10μg/ml的外泌体浓度作用强于5μg/ml,差异有统计学意义(P<0.05)。结论成功提取了hucMSCs及hucMSC-exo,hucMSC-exo可以以浓度依赖的方式促进mOPCs的增殖与成骨分化。

关键词：人脐带间充质干细胞外泌体成骨前体细胞增殖分化

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力学刺激在工程化心肌组织结构和功能改建中的作用研究

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组织工程与重建外科杂志 2006年第1期2卷 12-15页

作者：段翠密常海霞郭希民赵云山何文俊江红王常勇军事医学科学院基础医学研究所组织工程研究室北京100850

目的探索力学刺激在工程化心肌组织结构和功能改建中的作用。方法将分离并培养的新生大鼠心肌细胞与添加了Matrigel的液态Ⅰ型胶原混合,在环形槽中铸成环状心肌细胞/胶原条带。构建的条带在环形槽中培养7天后取出,在对其施加10%的静态... 详细信息

目的探索力学刺激在工程化心肌组织结构和功能改建中的作用。方法将分离并培养的新生大鼠心肌细胞与添加了Matrigel的液态Ⅰ型胶原混合,在环形槽中铸成环状心肌细胞/胶原条带。构建的条带在环形槽中培养7天后取出,在对其施加10%的静态拉伸力的情况下,再培养7天。通过光学显微镜、常规HE染色、免疫组织化学染色和透射电镜观察等对构建的组织工程化心肌组织进行评价。结果工程化心肌组织在力学刺激组的第2天可观察到局部跳动的区域,随时间的延长跳动的区域逐渐增多,强度逐渐增大,并最终形成一致跳动。而对照组未经力学拉伸刺激的工程化心肌组织则未能形成一致跳动。组织学和免疫组织化学染色研究结果显示,构建的工程化心肌组织经力学刺激,心肌细胞比较均匀地分布在整个条带中,且多数沿拉伸方向伸展,胞核呈长梭形,其形态类似于天然成熟心肌组织。透射电镜观察显示,条带中心肌细胞含有排列整齐的肌原纤维,并沿着细胞的长轴方向伸展,肌小节结构和Z带清晰可见。结论力学刺激可以更好地促进工程化心肌组织的结构和功能改建,使其更接近正常心肌组织。

关键词：力学刺激工程化心肌组织结构和功能改建大鼠心肌细胞免疫组织化学染色透射电镜观察跳动条带再培养环形槽构建光学显微镜显示区域力学拉伸拉伸方向胶原肌原纤维组织学

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