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Crosslinking strategies for preparation of extracellular matrix-derived cardiovascular scaffolds

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Regenerative Biomaterials 2014年第1期1卷 81-89页

作者： Bing Ma Xiaoya Wang Chengtie Wu Jiang Chang State Key Laboratory of High Performance Ceramics and Superfine Microstructure Shanghai Institute of CeramicsChinese Academy of Sciences1295 Dingxi RoadShanghai 200050People’s Republic of China

Heart valve and blood vessel replacement using artificial prostheses is an effective strategy for the treatment of cardiovascular disease at terminal stage.Natural extracellular matrix(ECM)-derived materials(decellularized allogeneic or xenogenic tissues)have received extensive attention as the cardiovascular scaffold.However,the bioprosthetic grafts usually far less durable and undergo calcification and progressive structural deterioration.Glutaraldehyde(GA)is a commonly used crosslinking agent for improving biocompatibility and durability of the natural scaffold materials.However,the nature ECM and GA-crosslinked materials may result in calcification and eventually lead to the transplant failure.Therefore,studies have been conducted to explore new crosslinking agents.In this review,we mainly focused on research progress of ECM-derived cardiovascular scaffolds and their crosslinking strategies.

关键词： extracellular matrix tissue engineering scaffold cross-linking agent calcification

Identification of SPP1 as an extracellular matrix signature for metastatic castration-resistant prostate cancer

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中国药理学与毒理学杂志 2019年第10期 760-761页

作者： PANG Xiao-cong XIE Ran ZHANG Zhuo LIU Qian-xin WU Shi-liang CUI Yi-min Department of Pharmacy Peking University First Hospital Department of Urology Peking University First Hospital

OBJECTIVE Resistance to androgen deprivation therapy(ADT) is the main challenge for advanced fatal prostate cancer(PCa), which can gradually develop into metastatic castration-resistant prostate cancer(m CRPC). However, the pathologic mechanisms of m CRPC are still far from clear. Given the high incidence and mortality related to m CRPC, understanding the causes and pathogenesis of this condition as well as identifying potential biomarkers are of great importance.METHODS In the research reported here, we integrated several gene expression profiles from hormone sensitive prostate cancer(HSPC) and m CRPC datasets to identify differentially expressed genes(DEGs), key biological pathways, and cellular components. K-nearest neighbor,logistic regression, naive Bayes, and random forest classifiers models were established to identify key regulated genes. Furthermore, organoid models were employed to validate the results in vitro. RESULTS We found that extracellular matrix(ECM) genes were significantly enriched, and further filtered them using Pearson correlation analysis and stepwise regression to find ECM signatures to differentiate between the HSPC and m CRPC phenotypes. Six ECM signatures were input into K-nearest neighbor, logistic regression, naive Bayes, and random forest classifiers models. Random forest algorithm with the six-gene prognostic signature showed best performance, which had high sensitivity and specificity for HSPC and m CRPC classification. Among the six ECM genes, SPP1 was identified as the key hub signature for PCa metastasis and drug resistance development; we found that both protein and m RNA expression levels of SPP1 were remarkably up-regulated in m CRPC compared with HSPC in organoid models and could regulate the androgen receptor signaling pathway. CONCLUSION Therefore, SPP1 is a potential novel biomarker and therapeutic target for m CRPC. Further understanding of the role of SPP1 in m CRPC development may help to explore effectively therapeutic approaches for the prevention and intervention

关键词： SPP1 castration-resistant prostate cancer extracellular matrix metastasis organoid model

Macrophage TGF-β1 and the Proapoptotic extracellular matrix Protein BIGH3 Induce Renal Cell Apoptosis in Prediabetic and Diabetic Conditions

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International Journal of Clinical Medicine 2016年第7期7卷 496-510页

作者： Robert J. Moritz Richard G. LeBaron Clyde F. Phelix Rajesha Rupaimoole Hong Seok Kim Andrew Tsin Reto Asmis Department of Biology University of Texas at San Antonio San Antonio USA Departments of Biochemistry and Clinical Laboratory Sciences School of Health Professions University of Texas Health Science Center at San Antonio San Antonio USA

Metabolically stressed kidney is in part characterized by infiltrating macrophages and macrophage-derived TGF-β1 that promote the synthesis of various ECM molecules. TGF-β1 strongly enhances the expression of the gene TGFBI that encodes a cell-adhesion class, proapoptotic ECM protein called BIGH3. We hypothesized that in a diabetic environment a relationship between infiltrating macrophages, macrophage-derived TGF-β1, and BIGH3 protein promotes renal cell death. To investigate this hypothesis, we used our mouse model of diabetic complications. Mice on a high-fat diet developed hypercholesterolemia, and exposure to streptozotocin rendered hypercholesterolemic mice diabetic. Immunohistochemical images show increased macrophage infiltration and BIGH3 protein in the kidney cortices of hypercholesterolemic and diabetic mice. Macrophages induced a two-fold increase in BIGH3 expression and an 86% increase in renal proximal tubule epithelial cell apoptosis. TGF-β1 antibody and TGF-β1 receptor chemical antagonist blocked macrophage-induced apoptosis. BIGH3 antibody completely blocked apoptosis that was induced by TGF-β1, and blocked apoptosis induced by exogenous recombinant BIGH3. These results uncover a distinctive interplay of macrophage-derived TGF-β1, BIGH3 protein, and apoptosis, and indicate that BIGH3 is central in a novel pathway that promotes diabetic nephropathy. Macrophage TGF-β1 and BIGH3 are identified as prediabetic biomarkers, and potential therapeutic targets for intervention in prediabetic and diabetic individuals.

关键词： TGF BIGH3 Macrophages Diabetes extracellular matrix

Harnessing the synergy of perfusable muscle flap matrix and adipose-derived stem cells for prevascularization and macrophage polarization to reconstruct volumetric muscle loss

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Bioactive Materials 2023年第4期22卷 588-614页

作者： Qixu Zhang Yulun Chiu Youbai Chen Yewen Wu Lina W.Dunne Rene D.Largo Edward I.Chang David M.Adelman Mark V.Schaverien Charles E.Butler Department of Plastic Surgery The University of Texas MD Anderson Cancer CenterHoustonTX77030USA Department of Melanoma Medical Oncology The University of Texas MD Anderson Cancer CenterHoustonTX77030USA Department of Plastic Surgery Chinese PLA General HospitalBeijing100853China

Muscle flaps must have a strong vascular network to support a large tissue volume and ensure successful engraftment.We developed porcine stomach musculofascial flap matrix(PDSF)comprising extracellular matrix(ECM)and intact vasculature.PDSF had a dominant vascular pedicle,microcirculatory vessels,a nerve network,well-retained 3-dimensional(3D)nanofibrous ECM structures,and no allo-or xenoantigenicity.In-depth proteomic analysis demonstrated that PDSF was composed of core matrisome proteins(e.g.,collagens,glycoproteins,proteoglycans,and ECM regulators)that,as shown by Gene Ontology term enrichment analysis,are functionally related to musculofascial biological processes.Moreover,PDSF􀀀human adipose-derived stem cell(hASC)synergy not only induced monocytes towards IL-10􀀀producing M2 macrophage polarization through the enhancement of hASCs’paracrine effect but also promoted the proliferation and interconnection of both human skeletal muscle myoblasts(HSMMs)and human umbilical vein endothelial cells(HUVECs)in static triculture conditions.Furthermore,PDSF was successfully prevascularized through a dynamic perfusion coculture of hASCs and HUVECs,which integrated with PDSF and induced the maturation of vascular networks in vitro.In a xenotransplantation model,PDSF demonstrated myoconductive and immunomodulatory properties associated with the predominance of M2 macrophages and regulatory T cells.In a volumetric muscle loss(VML)model,prevascularized PDSF augmented neovascularization and constructive remodeling,which was characterized by the predominant infiltration of M2 macrophages and significant musculofascial tissue formation.These results indicate that hASCs’integration with PDSF enhances the cells’dual function in immunomodulation and angiogenesis.Owing in part to this PDSF-hASC synergy,our platform shows promise for vascularized muscle flap engineering for VML reconstruction.

关键词： Muscle flap fabrication extracellular matrix Decellularization Vascularization Macrophage polarization Volumetric muscle loss

Shexiang Tongxin Dropping Pill Allieviates Heart Failure via Extracellula matrix-Receptor Interaction Pathways Based on RNA-Seq Transcriptomics and Experimental Studies

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Chinese Journal of Integrative Medicine 2023年第7期29卷 600-607页

作者： TAN Ya-fang FU Yu-han ZHANG Min-zhou Intensive Care Research Team of Traditional Chinese Medicine Guangdong Province Hospital of Chinese Medicinethe Second Affiliated Hospital of Guangzhou University of Chinese MedicineGuangzhou510006China School of Life Sciences and Biopharmaceutics Guangdong Pharmaceutical UniversityGuangzhou510006China

Objective:To investigate the protective mechanisms of Chinese medicine Shexiang Tongxin Dropping Pills(STDP) on heart failure(HF).Methods:Isoproterenol(ISO)-induced HF rat model and angiotensin Ⅱ(Ang Ⅱ)-induced neonatal rat cardiac fibroblast(CFs) model were used in the present study.HF rats were treated with and without STDP(3 g/kg).RNA-seq was performed to identify differentially expressed genes(DEGs).Cardiac function was evaluated by echocardiography.Hematoxylin and eosin and Masson’s stainings were taken to assess cardiac fibrosis.The levels of collagen Ⅰ(Col Ⅰ) and collagen Ⅲ(Col Ⅲ) were detected by immunohistochemical staining.CCK8 kit and transwell assay were implemented to test the CFs’ proliferative and migratory activity,respectively.The protein expressions of α-smooth muscle actin(α-SMA),matrix metalloproteinase-2(MMP-2),MMP-9,Col I,and Col Ⅲ were detected by Western blotting.Results:The results of RNA-seq analysis showed that STDP exerted its pharmacological effects on HF via multiple signaling pathways,such as the extracellular matrix(ECM)-receptor interaction,cell cycle,and B cell receptor interaction.Results from in vivo experiments demonstrated that STDP treatment reversed declines in cardiac function,inhibiting myocardial fibrosis,and reversing increases in Col Ⅰ and Col Ⅱ expression levels in the hearts of HF rats.Moreover,STDP(6,9 mg/mL) inhibited the proliferation and migration of CFs exposed to Ang Ⅱin vitro(P<0.05).The activation of collagen synthesis and myofibroblast generation were markedly suppressed by STDP,also the synthesis of MMP-2 and MMR-9,as well as ECM components Col Ⅰ,Col Ⅲ,and α-SMA were decreased in Ang Ⅱ-induced neonatal rats’ CFs.Conclusions:STDP had anti-fibrotic effects in HF,which might be caused by the modulation of ECM-receptor interaction pathways.Through the management of cardiac fibrosis,STOP may be a compelling candidate for improving prognosis of HF.

关键词： Shexiang Tongxin Dropping Pill heart failure RNA-seq extracellular matrix isoproterenol-induced

A decellularized nerve matrix scaffold inhibits neuroma formation in the stumps of transected peripheral nerve after peripheral nerve injury

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Neural Regeneration Research 2023年第3期18卷 664-670页

作者： Shuai Qiu Pei-Jun Deng Fu-Lin He Li-Wei Yan Zhe-Hui Tu Xiao-Lin Liu Da-Ping Quan Ying Bai Can-Bin Zheng Qing-Tang Zhu Department of Microsurgery and Orthopedic Trauma The First Affiliated HospitalSun Yat-sen UniversityGuangzhouGuangdong ProvinceChina Department of Orthopedic Surgery The Seventh Affiliated HospitalSun Yat-sen UniversityShenzhenGuangdong ProvinceChina Department of Pediatric Orthopedics Guangzhou Women and Children’s Medical CenterGuangzhou Medical UniversityGuangzhouGuangdong ProvinceChina Guangdong Provincial Peripheral Nerve Tissue Engineering and Technology Research Center GuangzhouGuangdong ProvinceChina Guangdong Province Engineering Laboratory for Soft Tissue Biofabrication GuangzhouGuangdong ProvinceChina School of Chemistry Sun Yat-sen UniversityGuangzhouGuangdong ProvinceChina School of Material Science and Engineering Sun Yat-sen UniversityGuangzhouGuangdong ProvinceChina

Traumatic painful neuroma is an intractable clinical disease characterized by improper extracellular matrix(ECM)deposition around the injury site.Studies have shown that the microstructure of natural nerves provides a suitable microenvironment for the nerve end to avoid abnormal hyperplasia and neuroma formation.In this study,we used a decellularized nerve matrix scaffold(DNM-S)to prevent against the formation of painful neuroma after sciatic nerve transection in rats.Our results showed that the DNM-S effectively reduced abnormal deposition of ECM,guided the regeneration and orderly arrangement of axon,and decreased the density of regenerated axons.The epineurium-perilemma barrier prevented the invasion of vascular muscular scar tissue,greatly reduced the invasion ofα-smooth muscle actin-positive myofibroblasts into nerve stumps,effectively inhibited scar formation,which guided nerve stumps to gradually transform into a benign tissue and reduced pain and autotomy behaviors in animals.These findings suggest that DNM-S-optimized neuroma microenvironment by ECM remodeling may be a promising strategy to prevent painful traumatic neuromas.

关键词： decellularized nerve matrix scaffold extracellular matrix fibrosis functional recovery microarchitecture microenvironment pain peripheral nerve tissue remodeling traumatic neuroma

Microtubes with gradient decellularized porcine sciatic nerve matrix from microfluidics for sciatic nerve regeneration

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Bioactive Materials 2023年第3期21卷 511-519页

作者： Binghui Jin Yunru Yu Xiangxiang Chen Yanhong Yang Yushan Xiong Young Jun Im Yuanjin Zhao Jian Xiao Oujiang Laboratory(Zhejiang Lab for Regenerative Medicine Vision and Brain Health)School of Pharmaceutical SciencesWenzhou Medical UniversityWenzhouZhejiang Province325035China Department of Rheumatology and Immunology Nanjing Drum Tower HospitalSchool of Biological Science and Medical EngineeringSoutheast UniversityNanjing210096China College of Pharmacy Chonnam National UniversityGwangju61186Republic of Korea Wenzhou Institute University of Chinese Academy of SciencesWenzhouZhejiang325001China

Long-range peripheral nerve defect is a severe and worldwide disease.With the increasing development of tissue engineering,the excellent ability of nerve extracellular matrix(ECM)in peripheral nerve injury(PNI)has been widely studied and verified.Here,we present a novel microtube that contains gradient decellularized porcine sciatic nerve ECM hydrogel(pDScNM-gel)from microfluidics for sciatic nerve regeneration.The pDScNM is confirmed to enhance cell proliferation and migration,and improve the axon growth of primary dorsal root ganglions(DRGs)in a concentration-related manner.These behaviors were also achieved when cells were co-cultured in a gradient pDScNM microtube.The in vivo sciatic nerve regeneration and functional recovery were also demonstrated by assembling the gradient pDScNM microtubes with a medical silicon tube.These results indicated that the microtubes with gradient pDScNM could act as a promising alternative for repairing peripheral nerve defects and showed great potential in clinical use.

关键词： Microfiber Peripheral nerve injury extracellular matrix Microfluidics Nerve regeneration

matrix stiffness modulates tip cell formation through the p-PXN-Rac1-YAP signaling axis

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Bioactive Materials 2022年第1期7卷 364-376页

作者： Yaru Guo Feng Mei Ying Huang Siqin Ma Yan Wei Xuehui Zhang Mingming Xu Ying He Boon Chin Heng Lili Chen Xuliang Deng Department of Geriatric Dentistry Peking University School and Hospital of StomatologyBeijing100081China Department of Stomatology Union HospitalTongji Medical CollegeHuazhong University of Science and TechnologyWuhan430022China Hubei Province Key Laboratory of Oral and Maxillofacial Development and Regeneration Wuhan430022China Department of Dental Materials Peking University School and Hospital of StomatologyBeijing100081PR China National Engineering Laboratory for Digital and Material Technology of Stomatology Beijing100081PR China Beijing Laboratory of Biomedical Materials Peking University School and Hospital of StomatologyBeijing100081PR China Central Laboratory Peking University School and Hospital of StomatologyBeijing100081PR China

Endothelial tip cell outgrowth of blood-vessel sprouts marks the initiation of angiogenesis which is critical in physiological and pathophysiological procedures.However,how mechanical characteristics of extracellular matrix(ECM)modulates tip cell formation has been largely neglected.In this study,we found enhanced CD31 expression in the stiffening outer layer of hepatocellular carcinoma than in surrounding soft tissues.Stiffened matrix promoted sprouting from endothelial cell(EC)spheroids and upregulated expressions of tip cell-enriched genes in vitro.Moreover,tip cells showed increased cellular stiffness,more actin cytoskeleton organization and enhanced YAP nuclear transfer than stalk and phalanx ECs.We further uncovered that substrate stiffness regulates FAK and Paxillin phosphorylation in focal adhesion of ECs promoting Rac1 transition from inactive to active state.YAP is subsequently activated and translocated into nucleus,leading to increased tip cell specification.p-Paxillin can also loosen the intercellular connection which also facilitates tip cell specification.Collectively our present study shows that matrix stiffness modulates tip cell formation through p-PXN-Rac1-YAP signaling axis,shedding light on the role of mechanotransduction in tip cell formation.This is of special significance in biomaterial design and treatment of some pathological situations.

关键词： Tip cells Mechanotransduction Stiffness Angiogenesis extracellular matrix

Function of cancer cell-derived extracellular matrix in tumor progression

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Journal of Cancer Metastasis and Treatment 2016年第1期2卷 357-364页

作者： Gao-Feng Xiong Ren Xu Markey Cancer Center LexingtonKY 40536USA Department of Pharmacology and Nutritional Sciences University of KentuckyLexingtonKY 40536USA

extracellular matrix(ECM)is an essential component of the tumor microenvironment.Cancer development and progression are associated with increased ECM deposition and crosslink.The chemical and physical signals elicited from ECM are necessary for cancer cell proliferation and invasion.It is well recognized that stromal cells are a major source of ECM proteins.However,recent studies showed that cancer cells are also an active and important component in ECM remodeling.Cancer cells deposit a significant amount of collagen,fibronectin,and tenascin C(TNC).Recent studies demonstrate that these cancer cell-derived ECM proteins enhance cancer cell survival and promote cancer cell colonization at distant sites.ECM-related enzymes and chaperone proteins,such as prolyl-4-hydroxylase,lysyl-hydroxylase,lysyl oxidase,and heat shock protein 47,are also highly expressed in cancer cells.Inhibition of these enzymes significantly reduces cancer growth,invasion,and metastasis.These factors suggest that the cancer cell-derived ECM is crucial for cancer progression and metastasis.Therefore,targeting these ECM proteins and ECM-related enzymes is a potential strategy for cancer treatment.

关键词： Tumor microenvironment extracellular matrix cancer progression metastasis