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Efficient induction of neural progenitor cells from human ESC/iPSCs on type i collagen

Science China(Life Sciences) 2021年第12期64卷 2100-2113页

作者： Pengfei Liu Shubin Chen Yaofeng Wang Xiaoming Chen Yiping Guo Chunhua Liu Haitao Wang Yifan Zhao Di Wu Yongli Shan Jian Zhang Chuman Wu Dongwei Li Yanmei Zhang Tiancheng Zhou Yaoyu Chen Xiaobo Liu Chenxu Li Lihui Wang Bei Jia Jie Liu Bo Feng Jinglei Cai Duanqing Pei CAS Key Laboratory of Regenerative Biology Guangdong Provincial Key Laboratory of Stem Cell and Regenerative MedicineGuangzhou Institutes of Biomedicine and HealthChinese Academy of SciencesGuangzhou 510530China Ambulatory Surgical Center The 2nd Clinical medical College(Shenzhen People’s Hospital)of Jinan UniversityThe 1st Affiliated Hospitals of Southern University of Science and TechnologyShenzhen 518020China integrated Chinese and Western Medicine Postdoctoral Research Station Jinan UniversityGuangzhou 510632China Bioland Laboratory Guangzhou Regenerative Medicine and Health Guangdong LaboratoryGuangzhou 510005China School of Biomedical Sciences The Chinese University of Hong KongHong Kong SARChina Centre for Regenerative Medicine and Health Hong Kong Institute of Science&InnovationChinese Academy of SciencesHong Kong SARChina Department of Regenerative Medicine School of Pharmaceutical ScienceJilin UniversityChangchun 130012China Department of Pathology Medical CollegeJinan UniversityGuangzhou 510632China The Center for Prenatal and Hereditary Disease Diagnosis Department of Obstetrics and GynecologyNanfang HospitalSouthern Medical UniversityGuangzhou 510515China Department of Obstetrics and Gynecology Nanfang HospitalSouthern Medical UniversityGuangzhou 510515China institute for Stem Cell and Regeneration Chinese Academy of SciencesBeijing 100101China

A stable,rapid and effective neural differentiation method is essential for the clinical applications of human embryonic stem cells(ESCs)or induced pluripotent stem cells(iPSCs)in treating neurological disorders and ***,we established a novel and robust monolayer differentiation method to produce functional neural progenitor cells(NPCs)from human ESC/iPSCs on type i *** derived cells not only displayed the requisite markers,but also behaved similarly to classic NPCs both in vitro and in *** transplantation into traumatic brain injury model,the derived NPCs facilitated recovery from *** also found that SMAD signaling stayed down throughout the differentiation process on type i collagen,and the pluripotent signals were rapidly downregulated along with raising up of neural early markers on the third ***,ATAC-seq data showed the related mediation of distinct transcriptome and global chromatin dynamics during NPC ***,our results thus provide a convenient way to generate NPCs from human ESC/iPSCs for neural diseases’treatment.

关键词： ESCs iPSCs NPCs neural differentiation type i collagen

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Effect of magnesium ions/type i collagen promote the biological behavior of osteoblasts and its mechanism

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Regenerative Biomaterials 2020年第1期7卷 53-61页

作者： Xiaojing Nie Xirao Sun Chengyue Wang Jingxin Yang Department of Prosthodontics The Second Affiliated Hospital of Jinzhou Medical UniversityJinzhou 121000China College of Robotics Beijing Union UniversityBeijing 100000China

type i collagen(Col i)is a main component of extracellular matrix(ECM).its safety,biocompatibility,hydrophilicity and pyrogen immunogenicity make it suitable for tissues engineering ***2t also control a myriad of cellular processes,including the bone development by enhancing the attachment and differentiation of osteoblasts and accelerating mineralization to enhance bone *** our studies,Mg2t bind collagen to promote the proliferation and differentiation of osteoblasts through the expression of integrins and downstream signaling *** order to clarify the biological behavior effect of 10mM Mg2t/Col i coating,we performed 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT),alkaline phosphatase(ALP),406-diamidino-2-phenylindole(DAPi),Alizarin red staining and Rhodamine B-isothiocyanate(RiTC)-labeled phalloidin experiments and found that 10mM Mg2t group,Col i-coating group,10mM Mg2t/Col i-coating group,respectively,promoted the proliferation and differentiation of osteoblasts,especially 10mM Mg2t/Col i-coating *** detected the mRNA expression of osteogenic-related genes(Runx2,ALP and OCN,OPN and BMP-2)and the protein expression of signaling pathway(integrin a2,integrin b1,FAK and ERK1/2),these results indicated that 10mM Mg2t/Col i coating play an critical role in up-regulating the MC3T3-E1 cells *** potential mechanisms of this specific performance may be through activating via integrin a2b1-FAK-ERK1/2 protein-coupled receptor pathway.

关键词： magnesium ion integrin type i collagen FAK/ERK

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Expressions of type i collagen, α2 integrin and β1 integrin in sclera of guinea pig with defocus myopia and inhibitory effects of bFGF on the formation of myopia

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international Journal of Ophthalmology(English edition) 2013年第1期6卷 54-58页

作者： Xiao-Dan Tian Yong-Xia Cheng Gui-Bo Liu Su-Fen Guo Chun-Lei Fan Li-Hui Zhan Yan-Chun Xu Department of Ophthalmology the First Hospital Affiliated to China Medical University Department of Ophthalmology the Fourth People's Hospital of Shenyang Department of Pathology Mudanjiang Medical University Department of Anatomy Mudanjiang Medical University

AiM:To investigate the expressions of type i collagen, α2 integrin and β1 integrin in the posterior sclera of guinea pigs with defocus myopia and whether basic fibroblast growth factor (bFGF) injection inhibits the formation and development of myopia by upregulating the expression of type i collagen, α2 integrin and β1 integrin. METHODS:After 14 days of treatment, the refractive state and axial length were measured and the levels of type i collagen, α2 integrin and β1 integrin were assayed in the posterior sclerae of groups of guinea pigs that wore a monocular-7D polymethylmethacrylate (PMMA) lens or had -7D lens wear followed by the peribulbar injection of Phosphate Buffer Solution (PBS) or bFGF. The untreated fellow eye served as a control. Guinea pigs with no treatment served as normal group. ·RESULTS:The results showed that 14 days of monocular defocus increased axial eye length and refraction, while bFGF delivery inhibited them markedly. Further, it was also found that the monocular-7D lens could decrease the levels of type i collagen, α2 integrin and β1 integrin expressions, while, unlike PBS, bFGF increased them significantly in comparison to contralateral control eyes and normal eyes. CONCLUSiON:bFGF can prevent the formation anddevelopment of defocus myopia by upregulating the expressions of type i collagen, α2 integrin and β1 integrin. Taken together, our results demonstrate that bFGF promotes sclera remodeling to prevent myopia in guinea pigs.

关键词： defocus myopia type i collagen α2 integrin β1 integrin bFGF

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The effect of Nifedipine on the expression of type i collagen in gingival fibroblasts

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Journal of Nanjing Medical University 2008年第2期22卷 92-95页

作者： Bei Li Weibin Sun Yong Ji Department of Periodontology Dental Research Institute Stomatological college Nanjing Medical University Nanjing 210029 Jiangsu Province China Department of Periodontology Stomatology hospital Nanjing University Nanjing 210029 Jiangsu Province China Department of Pathophysiology Nanjing Medical University Nanjing 210029Jiangsu Province China

Objective：To investigate the effect of nifedipine（calciumchannel blocker） on the expression of collagen in gingival fibroblasts invitro. Methods：Primarily gingival fibroblasts were cultured and incubated with various concentrations of nifedipine（108 μg/L, 360 μg/L and 1200 μg/L）for 5 days. Gingival fibroblasts were primarily cultured derived from nifedipine responders and nonresponders in the presence of 360 μg/L nifedipine. Enzyme-linked immunosorbent assay was used to evaluate the amount of type i collagen. Cell proliferation was measured by cell counting with evaluating MTT value. Results：The expressions of collagen and cell proliferation were significantly different among the high concentration groups and the others on the fifth day, especially higher in 360 μg/L and 1200 μg/L groups and also different among nifedipine responders and non-responders. Conclusion：The expression of collagen and cell proliferation may be concerned with the biological mechanism for gingival overgrowth.

关键词： fibroblast gingival hyperplasia type i collagen nifedipine

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Fabrication and Properties of Poly(vinylalcohol)-glycosaminoglycantype i collagen Composite Membrane as Tissue Regeneration Scaffolds

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Chinese Journal of Biomedical Engineering(English Edition) 2015年第4期24卷 139-146,159页

作者： Li Qin-hua LiN Dong-qing institute of Biomedical Engineering Jinan University Conmaster Eyeglasses Co.Ltd

The objective of this paper is to design a based on composite membrane with certain mechanical porous polyvinyl alcohol （PVA） strength and biocompatibilities serving as tissue regenerative scaffolds. PVA-glycosaminoglycan （GAG）-type i collagen （COL） composite membrane was fabricated by PVA with different molecular weight （Mw） and alcoholysis degree （AD） being blended with certain amounts of GAG and COL and dried at 38~C for 24 h. The water content of the composite membranes were from 61.9% to 95.1% and swelling ratio ranged from 123.6% to 621.7%. Scanning electron micro- scope （SEM） analysis proved that PVA-GAG-COL composite membrane has porous and homogenous structure. Biocompatibility test results showed that the composite membrane was nontoxic, which could promote adhesion and proliferation of fibroblasts on the com- posite membrane. in conclusion, PVA-GAG-COL composite membrane with high water content and swelling ratio, suitable mechanical strength and good biocompatibility, has potential in tissue engineering and regenerative medicine.

关键词： polyvinyl alcohol glycosaminoglycan type i collagen composite membrane tissue engineering

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Targeting collagen expression in alcoholic liver disease

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World Journal of Gastroenterology 2011年第20期17卷 2473-2481页

作者： Kyle J Thompson iain H McKillop Laura W Schrum Department of General Sur gery Carolinas Medical Center Liver Digestive and Metabolic Disorder LaboratoryCarolinas Medical Center

Alcoholic liver disease （ALD） is a leading cause of liver disease and liver-related deaths globally, particularly in developed nations. Liver fibrosis is a consequence of ALD and other chronic liver insults, which can progress to cirrhosis and hepatocellular carcinoma if left untreated. Liver fibrosis is characterized by accumulation of excess extracellular matrix components, including type i collagen, which disrupts liver microcirculation and leads to injury. To date, there is no therapy for the treatment of liver fibrosis; thus treatments that either prevent the accumulation of type i collagen or hasten its degradation are desirable. The focus of this review is to examine the regulation of type i collagen in fibrogenic cells of the liver and to discuss current advances in therapeutics to eliminate excessive collagen deposition.

关键词： type i collagen Fibrosis Extracellular matrix Hepatic stellate cell Alcohol Antioxidants Endoplasmic reticulum chaperones Matrix metalloproteinase microRNA

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Promotion of minTBP-1-PRGDN on the attachment,proliferation and collagen i synthesis of human keratocyte on titanium

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international Journal of Ophthalmology(English edition) 2014年第1期7卷 22-26页

作者： Xin-Yu Li Cai-Ni Ji Ling-Juan Xu Wei-Kun Hu Bin Zhou Gui-Gang Li Department of Ophthalmology Tongji HospitalTongji Medical CollegeHuazhong University of Science and Technology Department of Ophthalmology the Third Hospital of Wuhan Department of Stomatology Tongji HospitalHuazhong University of Science and Technology

AiM: To investigate the influence of minTBP-1-PRGDN on the attachment,proliferation and collagen i synthesis of human keratocyte on titanium(Ti) surface. · METHODS: The chimeric peptide RKLPDAPRGDN(minTBP-1-PRGDN) was synthesized by connecting RKLPDA(minTBP-1) to the N-terminal of PRGDN,the influence of minTBP-1-PRGDN on the attachment,proliferation and collagen i synthesis of human keratocyte on Ti surface were tested using PRGDN and minTBP-1 as controls. The keratocytes attached to the surface of Ti were either stained with FiTC-labeled phalloidin and viewed with fluorescence microscope or quantified with alamar Blue method. The proliferation of keratocytes on Ti were quantified with 3-(4,5-dim-ethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide uptaking methods. The secretion of type i collagen was determined using an ELiSA kit. ·RESULTS: The results showed that minTBP-1-PRGDN at a concentration of 100ng/mL was the most potent peptide to enhance the attachment of human keratocytes to the surface of Ti(1.40±0.03 folds,P =0.003),to promote the proliferation(1.26 ±0.05 folds,P =0.014) and the synthesis of type i collagen(1.530 ±0.128,P =0.008). MinTBP-1 at the same concentration could only promote the attachment(1.13±0.04 folds,P =0.020) and proliferation(1.15±0.06 folds,P =0.021),while PRGDN had no significant influence(P >0.05). ·CONCLUSiON: Our data show that the novel chimericpeptide minTBP-1-PRGDN could promote the attachment,proliferation and type i collagen synthesis of human keratocytes on the surface of titanium.

关键词： minTBP-1-PRGDN titanium cell attachment proliferation type i collagen

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Enhancement of meniscal repair in the avascular zone using connective tissue growth factor in a rabbit model

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Chinese Medical Journal 2011年第23期124卷 3968-3975页

作者： HE Wei LiU Yu-jie WANG Zhi-gang GUO Zi-kuan WANG Ming-xin WANG Ning Department of Orthopedic Surgery General Hospital of Chinese People＇s Liberation Army Beijing 100853 China

Background Connective tissue growth factor （CTGF） is a secreted protein containing several domains that mediate interactions with growth factors, integrins and extracellular matrix components. CTGF plays an important role in extracellular matrix production by its ability to mediate collagen deposition during wound healing. CTGF also induces neovascularization in vitro, suggesting a role in angiogenesis in vivo. We herein evaluated whether CTGF was required for extracellular matrix synthesis of meniscal fibrochondrocytes and/or angiogenesis during the repair of meniscal tears. Methods Meniscal fibrochondrocytes were isolated from the inner-i/2 of rabbit meniscus by trypsin collagenase treatment and further treated with 100 ng/ml CTGF in vitro. Characterization of fibrochondrocytes was identified by flow cytometry analyzing CD31, CD44, CD45 and CD105, and was further tested by type ii collagen immunocytochemistry. Changes in gene expression of meniscal fibrochondrocytes were monitored by quantitative real-time polymerase chain reaction. Histological sections prepared from a 3-mm portion of a longitudinal tearing defect in the middle of the rabbit meniscus were subjected to fluorescence-immunohistochemistry analysis at 1, 4 and 10 weeks following surgical treatment with 1.5 iJg of CTGF/fibrin-glue composites. Results Quantitative RT-PCR assay showed that types i and ii collagen and vascular endothelial growth factor mRNA expression in the 100 ng/ml CTGF group were remarkably enhanced as compared to levels in the no-dose group at 14 days （（2.38±0.63） fold, （2.96±0.87） fold, （2.14±0.56） fold, respectively）. Likewise, fluorescence-immunohistochemical analysis revealed that in the group implanted with CTGF-fibrin glue, types Ⅰand Ⅱcollagen, as well as the capillaries, completely filled the defect by 10 weeks, postoperatively. in contrast, only soft tissue repair occurred when PBS-fibrin glue was implanted. Conclusions These findings suggest that CTGF can significantly prom

关键词： connective tissue growth factor fibrochondrocyte meniscus vascular endothelial growth factor type i collagen

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Heme oxygenase-1 overexpression increases liver injury after bile duct ligation in rats

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World Journal of Gastroenterology 2007年第25期13卷 3478-3486页

作者： Matthias Froh Lars Conzelmann Peter Walbrun Susanne Netter Reiner Wiest Michael D Wheeler Mark Lehnert Takehiko Uesugi Jurgen Scholmerich Ronald G Thurman Department of internal Medicine University of Regensburg Germany Department of Urology University of Mainz Germany Department of Pharmacology University of North Carolina United States

AiM： To investigate the effects of heme oxygenase-1 （HO-1） against oxidant-induced injury caused by bile duct ligation （BDL）.METHODS： Either cobalt protoporphyrin （CoPP）, a HO-1 inducer, or saline were injected intraperitoneally in male SD-rats. Three days later, BDL or sham-operations were performed. Rats were sacrificed 3 wk after BDL and livers were harvested for histology. Fibrosis was evaluated by sirius red staining and image analysis. Alpha-smooth muscular actin, which indicates activation of stellate cells, was detected by immunohistochemical staining, and q/tokine and collagen- iα （Col- i α） mRNA expression was detected using RNase protection assays.RESULTS： Serum alanine transaminase increased 8-fold above normal levels one day after BDL. Surprisingly, enzyme release was not reduced in rats receiving CoPP. Liver fibrosis was evaluated 3 wk after BDL and the sirius red-positive area was found to be increased to about 7.8%. However, in CoPP pretreated rats sirius redpositive areas were increased to about 11.7% after BDL. collagen-1 α and TGF-β mRNA increased significantly by BDL. Again, this effect was increased by HO-1 overexpression.CONCLUSiON： Hepatic fibrosis due to BDL is not reduced by the HO-1 inducer CoPP. in contrast, HO-1 overexpression increases liver injury in rats under conditions of experimental chronic cholestasis.

关键词： Heme oxygenase-1 Bile duct ligation Chronic cholestasis Liver fibrosis Serum alaninetransaminase Transforming growth factor-13 Tumornecrosis factor- i ~ type i collagen

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Muscularis mucosae in desmoplastic stroma formation of early invasive rectal adenocarcinoma

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World Journal of Gastroenterology 2009年第39期15卷 4976-4979页

作者： Shinichi Ban Michio Shimizu Department of Pathology Saiseikai Kawaguchi General Hospital Department of Pathology Saitama Medical University International Medical Center

The origin of myofibroblasts or myofibroblastic cells in the desmoplastic stroma associated with carcinoma invasion has been controversial. in the early invasive area of a rectal adenocarcinoma reported here, an obvious transition between the muscularis mucosa and the bundles of eosinophilic stromal cells observed in the carcinomatous stroma was demonstrated both in morphology and in their cytoskeletal phenotype, which conceivably suggests that the smooth muscle cells of the muscularis mucosa could convert to the eosinophilic stromal cells, namely myofibroblasts. Moreover, type i procollagen was demonstrated in both protein and mRNA levels in the areas of eosinophilic stromal cells with a lesser degree of differentiated smooth muscle phenotype that showed a transition from the muscularis mucosa, implying that the myofibroblastic cells converted from smooth muscle cells of the muscularis mucosa could be responsible for type i collagen production. These findings suggest that the muscularis mucosae may not be a passive barrier through which colorectal carcinomas infiltrate into the submucosa, but may play an active role in the formation and remodeling of tumor stroma.

关键词： Rectal adenocarcinoma Muscularis mucosa Myofibroblast Cytoskeletal phenotype type i collagen

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