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Effects of adipose-derived mesenchymal stem cell Exosomes on Corneal Stromal Fibroblast Viability and Extracellular Matrix Synthesis

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Chinese Medical Journal 2018年第6期131卷 704-712页

作者： Ting Shen Qing-Qing Zheng Jiang Shen Qiu-Shi Li Xing-Hui Song Hong-Bo Luo Chao-Yang Hong Ke Yao Eye Institute of Zhejiang University Eye Center of Second Affiliated Hospital of Zhejiang University School of Medicine Hangzhou Zhejiang 310009 China Department of Ophthalmology Zhejiang Provincial People's Hospital and People's Hospital of Hangzhou Medical College Hangzhou Zhejiang 310014 China School of Ophthalmology and Optometry Wenzhou Medical University Wenzhou Zhejiang 325035 China Facility for Biochemistry and Molecular Medicine Core Facilities School of Medicine Zhejiang University Hangzhou Zhejiang 310058 China

Background： Corneal stromal cells （CSCs） are components of the corneal endothelial microenvironment that can be induced to form a functional tissue-engineered corneal endothelium. adipose-derived mesenchymal stem cells （ADSCs） have been reported as an important component of regenerative medicine and cell therapy for corneal stromal damage. We have demonstrated that the treatment with ADSCs leads to phenotypic changes in CSCs in vitro. However, the underlying mechanisms of such ADSC-induced changes in CSCs remain unclear. Methods： ADSCs and CSCs were isolated from New Zealand white rabbits and cultured in vitro. An Exosome Isolation Kit, Western blotting, and nanoparticle tracking analysis （NTA） were used to isolate and confirm the exosomes from ADSC culture medium. Meanwhile, the optimal exosome concentration and treatment time were selected. cell Counting Kit-8 and annexin V-fluorescein isothiocyanate/ propidium iodide assays were used to assess the effect of ADSC-derived exosomes on the proliferation and apoptosis of CSCs. To evaluate the effects ofADSC-derived exosomes on CSC invasion activity, Western blotting was used to detect the expression of matrix metalloproteinases （MMPs） and collagens. Results： ADSCs and CSCs were successfully isolated from New Zealand rabbits. The optimal concentration and treatment time of exosomes for the following study were 100 μg/ml and 96 h, respectively. NTA revealed that the ADSC-derived exosomes appeared as nanoparticles （40-200 nm）, and Western blotting confirmed positive expression of CD9, CDSI, flotillin-1, and HSP70 versus ADSC cytoplasmic proteins （all P〈 0.01 ）. ADSC-derived exosomes （50μg/ml and 100μg/ml） significantly promoted proliferation and inhibited apoptosis （mainly early apoptosis） of CSCs versus non-exosome-treated CSCs （all P 〈 0.05）. Interestingly, MMPs were downregulated and extracellular matrix （ECM）-related proteins including collagens and fibronectin were upregulated in the exosome-treated CSCs versus non-

关键词： adipose-derived mesenchymal stem cell Corneal Stromal cells Exosomes Extracellular Matrix Synthesis

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adipose-derived mesenchymal stem cells alleviate TNBS-induced colitis in rats by influencing intestinal epithelial cell regeneration, Wnt signaling, and T cell immunity

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World Journal of Gastroenterology 2020年第26期26卷 3750-3766页

作者： Jian-Guo Gao Mo-Sang Yu Meng-Meng Zhang Xue-Wei Gu Yue Ren Xin-Xin Zhou Dong Chen Tian-Lian Yan You-Ming Li Xi Jin Department of Gastroenterology The First Affiliated HospitalSchool of MedicineZhejiang UniversityHangzhou 310003Zhejiang ProvinceChina Department of Colorectal Surgery The First Affiliated HospitalSchool of MedicineZhejiang UniversityHangzhou 310003Zhejiang ProvinceChina

BACKGROUND Conventional Crohn’s disease(CD)treatments are supportive rather than curative and have serious side ***-derived mesenchymal stem cells(ADSCs)have been gradually applied to treat various *** therapeutic effect and underlying mechanism of ADSCs on CD are still not *** To investigate the effect of ADSC administration on CD and explore the potential *** Wistar rats were administered with 2,4,6-trinitrobenzene sulfonic acid(TNBS)to establish a rat model of CD,followed by tail injections of green fluorescent protein(GFP)-modified *** cytometry,qRT-PCR,and Western blot were used to detect changes in the Wnt signaling pathway,T cell subtypes,and their related *** The isolated cells showed the characteristics of ADSCs,including spindle-shaped morphology,high expression of CD29,CD44,and CD90,low expression of CD34 and CD45,and osteogenic/adipogenic *** therapy markedly reduced disease activity index and ameliorated colitis severity in the TNBS-induced rat model of ***,serum anti-sacchromyces cerevisiae antibody and panti-neutrophil cytoplasmic antibody levels were significantly reduced in ADSCtreated ***,the GFP-ADSCs were colocalized with intestinal epithelial cells(IECs)in the CD rat ***-ADSC delivery significantly antagonized TNBS-induced increased canonical Wnt pathway expression,decreased noncanonical Wnt signaling pathway expression,and increased apoptosis rates and protein level of cleaved caspase-3 in *** addition,ADSCs attenuated TNBS-induced abnormal inflammatory cytokine production,disturbed T cell subtypes,and their related markers in *** Successfully isolated ADSCs show therapeutic effects in CD by regulating IEC proliferation,the Wnt signaling pathway,and T cell immunity.

关键词： Crohn’s disease adipose-derived mesenchymal stem cell Intestinal epithelial cell Wnt pathway T cell Inflammation

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cell viability and extracellular matrix synthesis in a co-culture system of corneal stromal cells and adipose-derived mesenchymal stem cells

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International Journal of Ophthalmology(English edition) 2017年第5期10卷 670-678页

作者： Ting Shen Jiang Shen Qing-Qing Zheng Qiu-Shi Li Hai-Lan Zhao Lei Cui Chao-Yang Hong Zhejiang Provincial People's Hospital Hangzhou 310014Zhejiang Province China Beijing Shijitan Hospital Beijing 100050 China Wenzhou Medical University Wenzhou 325035 Zhejiang Province China

AIM：To investigate the impact of adipose-derived mesenchymal stem cells（ADSCs） on cell viability and extracellular matrix（ECM） synthesis of corneal stromal cells（CSCs）. METHODS：ADSCs and CSCs were obtained from the corneas of New Zealand white rabbits and indirectly cocultured in vitro. The proliferative capacity of CSCs in the different groups was assessed by CCK-8 assays. Annexin V-fluorescein isothiocyanate（FITC）/proliferation indices（PI） assays were used to detect the apoptosis of CSCs. The expression levels of matrix metalloproteinase（MMP）, such as MMP1, MMP2, MMP9, and collagens were also evaluated by Western blot. RESULTS：ADSCs significantly promoted proliferation and invasion of CSCs in the indirect co-culture assays. The co-cultural group displayed much higher ability of proliferation, especially under the co-culture conditions of ADSCs for 3d, compared with that CSCs cultured alone. The PI of CSCs in the co-culture system were increased approximately 3-8-fold compared with the control group. A significant change was observed in the proportions of cells at apoptosis（early and late） between the negative control group（6.34% and 2.06%） and the ADCSs-treated group（4.69% and 1.59%）. The expression levels of MMPs were down regulated in the co-culture models. Compared with the control group, the decrease intensities of MMP-1, MMP-2 and MMP-9 in CSCs/ADSCs group were observed, 3.90-fold, 1.09-fold and 3.03-fold, respectively. However, the increase intensities of collagen type（I, II, III, IV, and V） in CSCs were observed in CSCs/ADSCs group, 3.47-fold,4.30-fold, 2.35-fold, 2.55-fold and 2.43-fold, respectively, compared to that in the control group. The expressions of aldehyde dehydrogenase and fibronectin in CSCs were upregulated in the co-culture ***：ADSCs play a promotive role in CSCs＇ growth and invasion, which may be partially associated with MMPs decrease and collagens increase, resulting in a positive participation in the plasticity and ECM synt

关键词： adipose-derived mesenchymal stem cell corneal stromal cells extracellular matrix plasticity

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Advances of adipose-derived mesenchymal stem cells-based biomaterial scaffolds for oral and maxillofacial tissue engineering

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Bioactive Materials 2021年第8期6卷 2467-2478页

作者： Tong Liu Jia Xu Xun Pan Zhangfan Ding Hao Xie Xiaoyi Wang Huixu Xie State Key Laboratory of Oral Diseases National Clinical Research Center for Oral DiseasesDepartment of Head and Neck Oncology SurgeryWest China Hospital of StomatologySichuan UniversityChengdu610041China The Key Laboratory of Oral Biomedicine Jiangxi ProvinceSchool of StomatologyNanchang UniversityNanchang330006China General Surgery Department The Second Affiliated Hospital of Wannan Medical CollegeWuhuAnhui Province241000China

The management of oral and maxillofacial tissue defects caused by tumors,trauma,and congenital or acquired deformities has been a major challenge for surgeons over the last few *** tissue transplantation,the gold standard of tissue reconstruction,is a valid method for repairing the oral and maxillofacial functions and ***,several limitations hinder its clinical applications including complications of donor sites,limited tissue volume,and uncertain long-term ***-derived mesenchymal stem cells(ADMSCs)widely exist in adipose tissue and can be easily obtained through *** the bone marrow-derived mesenchymal stem cells(BMSCs),ADMSCs also have the multi-pluripotent potencies to differentiate into osteoblasts,chondrocytes,neurons,and ***,the multilineage capacity of ADMSCs makes them valuable for cell-based medical *** recent years,researchers have developed many candidates of ADMSCs-based biomaterial scaffolds to cater for the needs of oral and maxillofacial tissue engineering due to their superior *** review presents the advances and applications of ADMSCs-based biomaterial scaffolds,and explores their tissue engineering prospects in oral and maxillofacial reconstructions.

关键词： adipose-derived mesenchymal stem cell Biomaterial scaffold Oral and maxillofacial Tissue engineering

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ISOLATION AND INDUCTION OF DIFFERENTIATION OF SEINE adipose-derived mesenchymal stem cellS

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Journal of Otology 2014年第2期9卷 101-105页

作者： MA Yueying YUAN Shuolong ZHANG yue XU liangwei GUO Weiwei ZHAO Lidong ZHAI suoqiang YANG Shiming Department of Otolaryngology-Head and Neck Surgery Institute of OtolaryngologyChinese PLA General Hospital Department of Otolaryngology Head Neck SurgeryBeijing Friendship HospitalCapital Medical University Medical School of Nankai University

Objectives To establish a method for high yield mesenchymal stem cells collection, as well as a culture method for iden- tifying mesenchymal stem cells from the swine adipose-derived mesenchymal stem cell （ADMSC）. Methods Swine AD- MSCs were isolated from fat tissue with collagenase, followed by induction of differentiation to osteogenic, adipogenic and chondrogrnic cells. The survival curve of the ADMSC at the 37℃ and 38℃ were measured using WST-1 cell Proliferation Assay Reagent. Result ADMSCs isolated with collagenase from swine neck fat tissue generated a stable uniform appearance af- ter the second generation. The passage period was five days. ADMSC could differentiate into osteogenic, adipogenic or chon- drogrnic cells under different culture conditions. The highest growth rate was achieved at 38℃in this study. Conclusion Swine ADMSCs have the potential to differentiate into osteogenic, adipogenic or chondrogrnic cells, and they may be appropriate for transplantation for both research and clinical purpose.

关键词： adipose-derived mesenchymal stem cell Differentiation Transplantation Osteogenic cell, Adipogenic cell,Chondrogrnic cell

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A Review on the Biological Characteristics and Secretory Functions of adipose-derived mesenchymal stem cells

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Agricultural Science & Technology 2016年第6期17卷 1331-1335页

作者：刘雪婷支晓亮白春雨张明海关伟军东北林业大学野生动物资源学院黑龙江哈尔滨150040 中国农业科学院北京畜牧兽医研究所北京100193

adipose-derived mesenchymal stem cells （ADSCs） can be largely and easily obtained from a wide range of sources. Moreover, they have self-renewal ability, multi-differentiation potential, and an important role in immune regulation. They can secrete a variety of cytokines to regulate the in vivo micro-environment. Therefore, ADSCs are the ideal seed ceils for stem ceils application. This paper reviews the location, isolation, surface markers, proliferation, differentiation and other biological characteristics of ADSCs, as well as their secretory function and relative researches. ADSCs are expected to become excellent seed cells for cell therapy and tissue engineering through in-depth studies.

关键词： adipose-derived mesenchymal stem cell Surface markers Proliteration and differentiation Immune regulation Secretion

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Exosome/metformin-loaded self-healing conductive hydrogel rescues microvascular dysfunction and promotes chronic diabetic wound healing by inhibiting mitochondrial fission

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Bioactive Materials 2023年第8期26卷 323-336页

作者： Yue Zhang Meng Li Yunchuan Wang Fei Han Kuo Shen Liang Luo Yan Li Yanhui Jia Jian Zhang Weixia Cai Kejia Wang Ming Zhao Jing Wang Xiaowen Gao Chenyang Tian Baolin Guo Dahai Hu Department of Burns and Cutaneous Surgery Xijing HospitalFourth Military Medical University127 Changle West RoadXi’anShaanxi710032China State Key Laboratory for Mechanical Behavior of Materials And Frontier Institute of Science and TechnologyXi’an Jiaotong UniversityXi’an710049China Key Laboratory of Shaanxi Province for Craniofacial Precision Medicine Research College of StomatologyXi’an Jiaotong UniversityXi’an710049China

Chronic diabetic wounds remain a globally recognized clinical *** occur due to high concentrations of reactive oxygen species and vascular function disorders.A promising strategy for diabetic wound healing is the delivery of exosomes,comprising bioactive *** activates the vascular endothelial growth factor pathway,thereby improving angiogenesis in hyperglycemic ***,multifunctional hydrogels loaded with drugs and bioactive substances synergistically promote wound repair has been rarely reported,and the mechanism of their combinatorial effect of exosome and metformin in wound healing remains ***,we engineered dual-loaded hydrogels possessing tissue adhesive,antioxidant,self-healing and electrical conductivity properties,wherein 4-armed SH-PEG cross-links with Ag^(+),which minimizes damage to the loaded goods and investigated their mechanism of promotion effect for wound *** carbon nanotubes exhibiting good conductivity were also incorporated into the hydrogels to generate hydrogen bonds with the thiol group,creating a stable three-dimensional structure for exosome and metformin *** diabetic wound model of the present study suggests that the PEG/Ag/CNT-M+E hydrogel promotes wound healing by triggering cell proliferation and angiogenesis and relieving peritraumatic inflammation and vascular *** mechanism of the dual-loaded hydrogel involves reducing the level of reactive oxygen species by interfering with mitochondrial fission,thereby protecting F-actin homeostasis and alleviating microvascular ***,we propose a drug-bioactive substance combination therapy and provide a potential mechanism for developing vascular function-associated strategies for treating chronic diabetic wounds.

关键词： Dual-loaded hydrogels adipose-derived mesenchymal stem cell exosomes Metformin Angiogenesis Mitochondrial fission Chronic diabetic wound Wound healing

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Corneal stromal cells restoration in vitro by exosome derived from ADSCs

Corneal stromal cells restoration in vitro by exosome derive...

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2018年浙江省医学会激光医学分会学术年会暨浙江省医学会激光医学分会第二届学术年会

作者： Ting Shen Qingqing Zheng Jiang Shen Zhejiang Provincial people’s Hospital

adipose-derived mesenchymal stem cells(ADSCs) have been reported as an important component of the regenerative *** stromal cells(CSCs),as components of the corneal endothelial microenvironment,can be induced into a functional tissue-engineered corneal *** have demonstrated that ADSCs treatment leads to the phenotypic changes of CSCs in ***,the underlying mechanisms remain ***,we evaluated the effects of ADSCs-derived exosomes on CSCs in vitro using CCK-8 assays,Annexin V-FITC/PI assays,western blot and morphological *** tracking analysis technique(NTA) revealed ADSCsderived exosomes appeared as nanoparticles(40-200 nm),and western blot confirmed its positive expression of CD9,CD81,flotillin-1,and *** ADSCs exosome-treated CSCs showed significant difference in proliferation,apoptosis,especially the expression of extracellular matrix(ECM) related proteins,compared to the control ***-derived exosomes significantly promoted proliferation and inhibited the apoptosis of ***,MMPs were downregulated and collagens were upregulated in exosomes-treated *** of MMPs partially impeded the degradation of ECM,and increase of collagens contributed the ECM synthesis in *** findings indicate that ADSCs play an important role during CSCs’ ECM remodeling,which is closely related to phenotypic changes,partially through the secretion of exosomes.

关键词： adipose-derived mesenchymal stem cell corneal stromal cells extracellular matrix synthesis exosomes

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