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Genetic modification of miR-34a enhances efficacy of transplanted human dental pulp stem cells after ischemic stroke

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Neural Regeneration Research 2023年第9期18卷 2029-2036页

作者： Jianfeng Wang Peibang He Qi Tian Yu Luo Yan He Chengli Liu Pian Gong Yujia Guo Qingsong Ye Mingchang Li department of Neurosurgery Renmin Hospital of Wuhan UniversityWuhanHubei ProvinceChina Center of regenerative Medicine Renmin Hospital of Wuhan UniversityWuhanHubei ProvinceChina Institute of regenerative and Translational Medicine Tianyou HospitalWuhan University of Science and TechnologyWuhanHubei ProvinceChina

Human dental pulp stem cells(hDPSCs) promote recovery after ischemic stro ke;however,the therapeutic efficacy is limited by the poor survival of transplanted *** in vitro expe riments in the present study,we used oxygen-glucose deprivation/reoxygenation in hDPSCs to mimic cell damage induced by ischemia/*** found that miRNA-34a-5p(miR-34a) was elevated under oxygen-glucose deprivation/reoxygenation conditions in *** of miR-34a facilitated the prolife ration and antioxidant capacity and reduced the apoptosis of *** r,dual-luciferase reporter gene assay showed WNT1and SIRT1 as the targets of *** miR-34a knockdown cell lines,WNT1 suppression reduced cell prolife ration,and SIRT1 suppression decreased the antioxidant *** r,these results indicated that miR-34a regulates cell prolife ration and antioxidant stress via targeting WNT1 and SIRT1,*** in vivo expe riments,we injected genetically modified hDPSCs(anti34a-hDPSCs) into the brains of *** found that anti34a-hDPSCs significantly inhibited apoptosis,reduced cerebral edema and cerebral infarct volume,and improved motor function in *** study provides new insights into the molecular mechanism of the cell prolife ration and antioxidant capacity of hDPSCs,and suggests a potential gene that can be targeted to improve the survival rate and efficacy of transplanted hDPSCs in brain after ischemic stroke.

关键词： antioxidant capacity HO-1 human dental pulp stem cells ischemic stroke miR-34a Nrf2 proliferation SIRT1 WNT1 β-catenin

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Eicosane, pentadecane and palmitic acid: The effects in in vitro wound healing studies

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Asian Pacific Journal of Tropical Biomedicine 2018年第10期8卷 490-499页

作者： Xin Qi Chuah Patrick Nwabueze Okechukwu Farahnaz Amini Swee Sen Teo department of Biotechnology Faculty of Applied SciencesUCSI University School of Healthy Aging Medical Aesthetics and Regenerative MedicineUCSI University

Objective: To examine the wound healing properties of eicosane, pentadecane and palmitic acid by evaluating in term of anti-microbial, anti-inflammatory, proliferation, migration and collagen synthesis. Methods: Anti-microbial activities of Staphylococcus aureus, Bacillus subtilis, Escherichia coli and Pseudomonas aeruginosa were evaluated by carrying out disk diffusion and agar well diffusion methods. Growth rate of tested bacteria was also evaluated for 8 h in conjunction with the sample drugs. Besides, U937 cell lines were used as model study for realtime mRNA genes expression studies of TNF-毩 and IL-12 under the treatment. Proliferation, migration and collagen content synthesis were carried out on human dermal fibroblast. Results: None of the sample drugs possessed significant inhibition of bacteria tested in this study both in disk diffusion and agar well diffusion methods. In contrary, significantly low expressed mRNA gene expression levels of TNF-毩 and IL-12 were found under the treatment of respective drugs. Meanwhile in proliferation, migration and hydroxyproline content analysis, all the sample drugs showed no significant positive stimulation. Conclusions: This study therefore explains that apart from their potential in downregulating pro-inflammatory cytokines, these three compounds which were examined individually may not be good candidates in promoting wound healing.

关键词： Eicosane Pentadecane Palmitic acid Wound healing TNF-α IL-12

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Effects of Rosmarinic Acid on DNA Integrity and H19 Differentially Methylated Region Methylation Levels in Human Sperm Preserved by Freeze-Drying

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Reproductive and Developmental Medicine 2021年第1期5卷 9-14页

作者： Yi-Yu Wang Wei-Jie Zhu department of Developmental and regenerative biology College of Life Science and TechnologyJinan UniversityGuangzhou 510632China

Original Article Effects of Rosmarinic Acid on DNA Integrity and H19 Differentially Methylated Region Methylation Levels in Human Sperm Preserved by Freeze-Drying Wang Yi-Yu,Zhu Wei-Jie Published 2021-03-25 Cite as Reprod Dev Med,2021,05 Reprod Dev Med,2021,05(1):***:10.4103/2096-2924.309790 Abstract Objective:To investigate the effects of rosmarinic acid(RA)on the DNA integrity and methylation levels of the H19 differentially methylated region(DMR)of freeze-dried human sperm after 1 week and 6 months of storage at 4℃.Methods:Semen samples from 15 healthy normospermic donors were used in this *** samples were divided into five groups,including the control group with fresh sperm and four experimental groups with freeze-dried sperm(1-week storage with EGTA buffer solution,Group A;1-week storage with EGTA buffer solution containing 105μmol/L RA,Group B;6-month storage with EGTA buffer solution,Group C;and 6-month storage with EGTA buffer solution containing 105μmol/L RA,Group D).DNA integrity was evaluated using the sperm chromatin dispersion test.H19 DMR methylation levels were detected by bisulfite sequencing polymerase chain ***:After 1 week of storage,no differences in sperm DNA integrity were observed among Groups A,B,and controls(P>0.05).After 6 months of storage,the sperm DNA integrity of Group D did not change significantly compared with that of the control group(P>0.05),whereas that of Group C decreased significantly(P0.05).Conclusions:The DNA integrity of freeze-dried human sperm can be effectively protected by adding RA within 6 months,and the H19 DMR methylation level of human sperm can be maintained for 6 months after freeze-drying.

关键词： DNA Integrity DNA Methylation Freeze-Drying Human Sperm Rosmarinic Acid

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p21/Zbtb18 repress the expression of cKit to regulate the self-renewal of hematopoietic stem cells

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Protein & Cell 2024年第11期15卷 840-857页

作者： Nini Wang Shangda Yang Yu Li Fanglin Gou Yanling Lv Xiangnan Zhao Yifei Wang Chang Xu Bin Zhou Fang Dong Zhenyu Ju Tao Cheng Hui Cheng State Key Laboratory of Experimental Hematology National Clinical Research Center for Blood DiseasesHaihe Laboratory of Cell EcosystemInstitute of Hematology&Blood Diseases HospitalChinese Academy of Medical Sciences&Peking Union Medical CollegeTianjin 300020China CAMS Center for Stem Cell Medicine PUMC Department of Stem Cell and Regenerative MedicineTianjin 300020China The Province and Ministry Co-sponsored Collaborative Innovation Center for Medical Epigenetics Key Laboratory of Immune Microenvironment and Disease(Ministry of Education)Department of Cell BiologyTianjin Medical UniversityTianjin 300270China State Key Laboratory of Cell biology Shanghai Institute of Biochemistry and Cell BiologyCenter for Excellence in Molecular Cell ScienceChinese Academy of SciencesUniversity of Chinese Academy of SciencesShanghai 200031China Key Laboratory of regenerative Medicine of Ministry of Education Institute of Aging and Regenerative MedicineCollege of Life Science and TechnologyJinan UniversityGuangzhou 510632China

The maintenance of hematopoietic stem cells(HSCs)is a complex process involving numerous cell-extrinsic and-intrinsic *** first member of the cyclin-dependent kinase family of inhibitors to be identified,p21,has been reported to perform a wide range of critical biological functions,including cell cycle regulation,transcription,differentiation,and so *** the previous inconsistent results regarding the functions of p21 in HSCs in a p21-knockout mouse model,we employed p21-tdTomato(tdT)mice to further elucidate its role in HSCs during *** results showed that p21-tdT+HSCs exhibited increased self-renewal capacity compared to p21-tdT−***18,a transcriptional repressor,was upregulated in p21-tdT+HSCs,and its knockdown significantly impaired the reconstitution capability of ***,p21 interacted with ZBTB18 to co-repress the expression of cKit in HSCs and thus regulated the self-renewal of *** data provide novel insights into the physiological role and mechanisms of p21 in HSCs during homeostasis independent of its conventional role as a cell cycle inhibitor.

关键词： hematopoietic stem cells self-renewal p21 Zbtb18 cKit

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Adult stem-like cells in kidney

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World Journal of Stem Cells 2015年第2期7卷 490-494页

作者： Keiichi Hishikawa Osamu Takase Masahiro Yoshikawa Taro Tsujimura Masaomi Nangaku Tsuyoshi Takato department of Advanced Nephrology and regenerative Medicine the University of Tokyo department of Oral and Maxillofacial Surgery Graduate School of MedicineUniversity of Tokyo

Human pluripotent cells are promising for treatment for kidney diseases, but the protocols for derivation of kidney cell types are still controversial. Kidney tissue regeneration is well confirmed in several lower vertebrates such as fish, and the repair of nephrons after tubular damages is commonly observed after renal injury. Even in adult mammal kidney, renal progenitorcell or system is reportedly presents suggesting that adult stem-like cells in kidney can be practical clinical targets for kidney diseases. However, it is still unclear if kidney stem cells or stem-like cells exist or not. In general, stemness is defined by several factors such as self-renewal capacity, multi-lineage potency and characteristic gene expression profiles. The definite use of stemness may be obstacle to understand kidney regeneration, and here we describe the recent broad findings of kidney regeneration and the cells that contribute regeneration.

关键词： Gene therapy Hematopoietic stem cells In vivo selection Chemical Inducer of Dimerization Chemo-selection Lentivirus

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Caffeic acid and protocatechuic acid modulate Nrf2 and inhibit Ehrlich ascites carcinomas in mice

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Asian Pacific Journal of Tropical Biomedicine 2021年第6期11卷 244-253页

作者： Venugopal R.Bovilla Preethi G.Anantharaju Sireesh Dornadula Prashanthkumar M.Veeresh Mahadevaswamy G.Kuruburu Vidya G.Bettada Kunka Mohanram Ramkumar SubbaRao V.Madhunapantula Center of Excellence in Molecular biology and regenerative Medicine Department of BiochemistryJSS Medical CollegeJSS Academy of Higher Education&ResearchMysore-570015KarnatakaIndia SRM Research Institute Department of BiotechnologySchool of BioengineeringSRM Institute of Science and TechnologyKattankulathur-603203Tamil NaduIndia

Objective:To assess the nuclear factor-erythroid 2-related factor-2(Nrf2)modulatory effect of caffeic acid and protocatechuic acid and determine the anti-tumor activity of these phenolic compounds against Ehrlich ascites carcinoma growth in ***:Antioxidant activity of protocatechuic acid and caffeic acid was assessed using ferric reducing antioxidant power(FRAP)and 2,2-diphenyl-1-picrylhydrazyl(DPPH).Nrf2 activation potential of phenolic compounds was tested by quantitative realtime polymerase chain reaction,and luciferase complementation reporter *** vivo efficacy was tested using the Ehrlich ascites carcinoma ***:FRAP and DPPH radical scavenging assays showed that caffeic acid and protocatechuic acid were more potent compared with cinnamic acid and benzoic *** complementation reporter assays identified caffeic acid and protocatechuic acid as the activators of *** caffeic acid and protocatechuic acid upregulated the expression of Nrf2 target genes heme oxygenase-1(HO-1),glutamate-cysteine ligase catalytic subunit(GCLC),and glutamate-cysteine ligase modifier subunit(GCLM)and the activity of NAD(P)H:quinone oxidoreductase 1(NQO1)when tested on HCT-116 cells using a cell-based assay system at 9 *** addition,intraperitoneal administration of caffeic acid and protocatechuic acid to Ehrlich ascites carcinoma bearing mice suppressed tumor growth and ***:Caffeic acid and protocatechuic acid can modulate Nrf2 and inhibit Ehrlich ascites carcinoma cells.

关键词： Ehrlich ascites carcinomas Nrf2 Protocatechuic acid Caffeic acid NQO1 activity

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Renal progenitors: Roles in kidney disease and regeneration

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World Journal of Stem Cells 2016年第11期8卷 367-375页

作者： Brooke E Chambers Rebecca A Wingert department of Biological Sciences Center for Zebrafish Research Center for Stem Cells and Regenerative Medicine University of Notre Dame

Kidney disease is a devastating condition that affects millions of people worldwide, and its prevalence predicted to significantly increase. The kidney is complex organ encompassing many diverse cell type organized in a elaborate tissue architecture, makin regeneration a challenging feat. In recent years, there ha been a surge in the field of stem cell research to develo regenerative therapies for various organ systems. Here we review some recent progressions in characterizing th role of renal progenitors in development, regeneration and kidney disease in mammals. We also discuss how the zebrafish provides a unique experimental anima model that can provide a greater molecular and genet understanding of renal progenitors, which may contribut to the development of potential regenerative therapies fo human renal afflictions.

关键词： Kidney Renal progenitor Nephrogenesis Development Nephron Regeneration Zebrafish Parietal epithelial cell Tubular progenitor cell

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Fargeted elimination of mutant mitochondria DNA in MELAS-iPSCs by mitoTALENs

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Protein & Cell 2018年第3期9卷 283-297页

作者： Yi Yang Han Wu Xiangjin Kang Yanhui Liang Ting Lan Tianjie Li Tao Tan Jiangyun Peng Quanjun Zhang Geng An Yali Liu Qian Yu Zhenglai Ma Ying Lian Boon Seng Soh Qingfeng Chen Ping Liu Yaoyong Chen Xiaofang Sun Rong Li Xiumei Zhen Yang Yu Xiaoping Li Yong Fan Key Laboratory for Major Obstetric Diseases of Guangdong Province Key Laboratory of Reproduction and Genetics of Guangdong Higher Education Institutes The Third Affiliated Hospital of Guangzhou Medical University Guangzhou 510150 China Key Laboratory of regenerative biology of the Chinese Academy of Sciences and Guangdong Provincial Key Laboratory of Stem Cells and regenerative Medicine South China Institute for Stem Cell Biology and Regenerative Medicine Guangzhou Institutes of Biomedicine and Health Chinese Academy of Sciences Guangzhou 510530 China Center of Reproductive Medicine Department of Obstetrics and Gynecology Peking University Third Hospital Beijing 100191 China Yunnan Key Laboratory of Primate Biomedical Research Institute of Primate Translational Medicine Kunming University of Science and Technology Kunming 650500 China Disease Modeling and Therapeutics Laboratory A'STAR Institute of Molecular and Cell Biology 61 Biopolis Drive Proteos Singapore 138673 Singapore department of Biological Sciences National University of Singapore Singapore 117543 Singapore

Mitochondrial diseases are maternally inherited hetero- geneous disorders that are primarily caused by mitochondrial DNA （mtDNA） mutations. Depending on the ratio of mutant to wild-type mtDNA, known as heteroplasmy, mitochondrial defects can result in a wide spectrum of clinical manifestations. Mitochondria-targeted endonucleases provide an alternative avenue for treating mitochondrial disorders via targeted destruc- tion of the mutant mtDNA and induction of heteroplasmic shifting. Here, we generated mitochondrial disease patient-specific induced pluripotent stem cells （MiPSCs） that harbored a high proportion of m.3243A〉G mtDNA mutations and caused mitochondrial encephalomyopathy and stroke-like episodes （MELAS）. We engineered mitochondrial-targeted transcription activator-like effector nucleases （mitoTALENs） and successfully eliminated the m.3243A〉G mutation in MiPSCs. Off-target mutagenesis was not detected in the targeted MiPSC clones. Utilizing a dual fluorescence iPSC reporter cell line expressing a 3243G mutant mtDNA sequence in the nuclear genome, mitoTALENs displayed a significantly limited ability to target the nuclear genome compared with nuclear-localized TALENs. Moreover, genetically rescued MiPSCs displayed normal mitochondrial respiration and energy production. Moreover, neuronal progenitor cells differentiated from the rescued MiPSCs also demonstrated normal metabolic profiles. Further- more, we successfully achieved reduction in the human m.3243A〉G mtDNA mutation in porcine oocytes via injection of mitoTALEN mRNA. Our study shows the great potential for using mitoTALENs for specific targeting of mutant mtDNA both in iPSCs and mammalian oocytes, which not only provides a new avenue for studying mitochondrial biology and disease but also suggests a potential therapeutic approach for the treatment of mitochondrial disease, as well as the prevention of germline transmission of mutant mtDNA.

关键词： mitochondria iPSCs TALEN MELAS

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Selective serotonin re-uptake inhibitor sertraline inhibits bone healing in a calvarial defect model

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International Journal of Oral Science 2018年第4期10卷 212-222页

作者： R.Nicole Howie Samuel Herberg Emily Durham Zachary Grey Grace Bennfors Mohammed Elsalanty Amanda C.LaRue William D.Hill James J.Cray Oral Health Sciences Medical University of South CarolinaCharlestonSCUSA Biomedical Engineering Case Western Reserve UniversityClevelandOHUSA Cellular biology and Anatomy Augusta UniversityAugustaGAUSA Oral biology Augusta UniversityAugustaGAUSA Orthopaedic Surgery Augusta UniversityAugustaGAUSA Pathology and Laboratory Medicine Medical University of South CarolinaCharlestonSCUSA Institute for regenerative and Reparative Medicine Augusta UniversityAugustaGAUSA Research Service of the Ralph H Johnson VA Medical Center CharlestonSCUSA Charlie Norwood VA Medical Center AugustaGAUSA department of regenerative Medicine and Cellular biology CharlestonSCUSA Wake Forest Institute for regenerative Medicine Wake Forest School of MedicineWinston-SalemNCUSA Division of Anatomy College of MedicineOhio State UniversityColumbusOHUSA

Bone wound healing is a highly dynamic and precisely controlled process through which damaged bone undergoes repair and complete regeneration. External factors can alter this process, leading to delayed or failed bone wound healing. The findings of recent studies suggest that the use of selective serotonin reuptake inhibitors(SSRIs) can reduce bone mass, precipitate osteoporotic fractures and increase the rate of dental implant failure. With 10% of Americans prescribed antidepressants, the potential of SSRIs to impair bone healing may adversely affect millions of patients’ ability to heal after sustaining trauma. Here, we investigate the effect of the SSRI sertraline on bone healing through pre-treatment with(10 mg·kg-1sertraline in drinking water, n = 26) or without(control, n = 30) SSRI followed by the creation of a 5-mm calvarial defect. Animals were randomized into three surgical groups:(a) empty/sham,(b) implanted with a DermaMatrix scaffold soak-loaded with sterile PBS or(c) DermaMatrix soak-loaded with542.5 ng BMP2. SSRI exposure continued until sacrifice in the exposed groups at 4 weeks after surgery. Sertraline exposure resulted in decreased bone healing with significant decreases in trabecular thickness, trabecular number and osteoclast dysfunction while significantly increasing mature collagen fiber formation. These findings indicate that sertraline exposure can impair bone wound healing through disruption of bone repair and regeneration while promoting or defaulting to scar formation within the defect site.

关键词： Selective serotonin re-uptake inhibitor sertraline inhibits bone healing in a calvarial defect model

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Magnetically responsive nanofibrous ceramic scaffolds for on-demand motion and drug delivery

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Bioactive Materials 2022年第9期7卷 372-381页

作者： Yonggang Zhang Jiaping Li Pamela Habibovic department of Instructive Biomaterials Engineering Maastricht UniversityMERLN Institute for Technology-Inspired Regenerative MedicineUniversiteitssingel 406229 ERMaastrichtthe Netherlands department of Complex Tissue Regeneration Maastricht UniversityMERLN Institute for Technology-Inspired Regenerative MedicineUniversiteitssingel 406229 ERMaastrichtthe Netherlands

Smart biomaterials,featuring not only bioactivity,but also dynamic responsiveness to external stimuli,are desired for biomedical applications,such as regenerative medicine,and hold great potential to expand the boundaries of the modern clinical ***,a magnetically responsive three-dimensional scaffold with sandwich structure is developed by using hydroxyapatite(HA)nanowires and ferrosoferric oxide(Fe_(3)O_(4))nanoparticles as building *** magnetic HA/Fe_(3)O_(4) scaffold is fully inorganic in nature,but shows polymeric hydrogel-like characteristics including a 3D fibrous network that is highly porous(>99.7%free volume),deformable(50%deformation)and elastic,and tunable *** magnetic HA/Fe3O4 scaffold has been shown to execute multimodal motion upon exposure to an external magnetic field including shape transformation,rolling and *** addition,we have demonstrated that the magnetic scaffold can serve as a smart carrier for remotely controlled,on-demand delivery of compounds including an organic dye and a ***,the magnetic scaffold has exhibited good biocompatibility,supporting the attachment and proliferation of human mesenchymal stromal cells,thereby showing great potential as smart biomaterials for a variety of biomedical applications.

关键词： Hydroxyapatite Magnetic Responsive Drug delivery Cell culture

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