BACKGROUND Cartilage defects are some of the most common causes of arthritis.Cartilage lesions caused by inflammation,trauma or degenerative disease normally result in osteochondral defects.Previous studies have shown...BACKGROUND Cartilage defects are some of the most common causes of arthritis.Cartilage lesions caused by inflammation,trauma or degenerative disease normally result in osteochondral defects.Previous studies have shown that decellularized extracellular matrix(ECM)derived from autologous,allogenic,or xenogeneic mesenchymal stromal cells(MSCs)can effectively restore osteochondral integrity.AIM To determine whether the decellularized ECM of antler reserve mesenchymal cells(RMCs),a xenogeneic material from antler stem cells,is superior to the currently available treatments for osteochondral defects.METHODS We isolated the RMCs from a 60-d-old sika deer antler and cultured them in vitro to 70%confluence;50 mg/mL L-ascorbic acid was then added to the medium to stimulate ECM deposition.Decellularized sheets of adipocyte-derived MSCs(aMSCs)and antlerogenic periosteal cells(another type of antler stem cells)were used as the controls.Three weeks after ascorbic acid stimulation,the ECM sheets were harvested and applied to the osteochondral defects in rat knee joints.RESULTS The defects were successfully repaired by applying the ECM-sheets.The highest quality of repair was achieved in the RMC-ECM group both in vitro(including cell attachment and proliferation),and in vivo(including the simultaneous regeneration of well-vascularized subchondral bone and avascular articular hyaline cartilage integrated with surrounding native tissues).Notably,the antler-stem-cell-derived ECM(xenogeneic)performed better than the aMSC-ECM(allogenic),while the ECM of the active antler stem cells was superior to that of the quiescent antler stem cells.CONCLUSION Decellularized xenogeneic ECM derived from the antler stem cell,particularly the active form(RMC-ECM),can achieve high quality repair/reconstruction of osteochondral defects,suggesting that selection of decellularized ECM for such repair should be focused more on bioactivity rather than kinship.展开更多
An outbreak of coronavirus disease 2019(COVID-19)has spread globally,with over 500 million cases and 6 million deaths to date.COVID-19 is associated with a systemic inflammatory response and abnormalities of the extra...An outbreak of coronavirus disease 2019(COVID-19)has spread globally,with over 500 million cases and 6 million deaths to date.COVID-19 is associated with a systemic inflammatory response and abnormalities of the extracellular matrix(ECM),which is also involved in inflammatory storms.Upon viral infection,ECM proteins are involved in the recruitment of inflammatory cells and interference with target organ metabolism,including in the lungs.Additionally,serum biomarkers of ECM turnover are associated with the severity of COVID-19 and may serve as potential targets.Consequently,understanding the expression and function of ECM,particularly of the lung,during severe acute respiratory syndrome of the coronavirus 2 infection would provide valuable insights into the mechanisms of COVID-19 progression.In this review,we summarize the current findings on ECM,such as hyaluronic acid,matrix metalloproteinases,and collagen,which are linked to the severity and inflammation of COVID-19.Some drugs targeting the extracellular surface have been effective.In the future,these ECM findings could provide novel perspectives on the pathogenesis and treatment of COVID-19.展开更多
The bone extracellular matrix(ECM) contains minerals deposited on highly crosslinked collagen fibrils and hundreds of noncollagenous proteins. Some of these proteins are key to the regulation of bone formation and reg...The bone extracellular matrix(ECM) contains minerals deposited on highly crosslinked collagen fibrils and hundreds of noncollagenous proteins. Some of these proteins are key to the regulation of bone formation and regeneration via signaling pathways,and play important regulatory and structural roles. However, the complete list of bone extracellular matrix proteins, their roles, and the extent of individual and cross-species variations have not been fully captured in both humans and model organisms. Here, we introduce the most comprehensive resource of bone extracellular matrix(ECM) proteins that can be used in research fields such as bone regeneration, osteoporosis, and mechanobiology. The Phylobone database(available at https://phylobone.com) includes 255proteins potentially expressed in the bone extracellular matrix(ECM) of humans and 30 species of vertebrates. A bioinformatics pipeline was used to identify the evolutionary relationships of bone ECM proteins. The analysis facilitated the identification of potential model organisms to study the molecular mechanisms of bone regeneration. A network analysis showed high connectivity of bone ECM proteins. A total of 214 functional protein domains were identified, including collagen and the domains involved in bone formation and resorption. Information from public drug repositories was used to identify potential repurposing of existing drugs. The Phylobone database provides a platform to study bone regeneration and osteoporosis in light of(biological) evolution,and will substantially contribute to the identification of molecular mechanisms and drug targets.展开更多
Carious lesions are bacteria-caused destructions of the mineralised dental tissues,marked by the simultaneous activation of immune responses and regenerative events within the soft dental pulp tissue.While major molec...Carious lesions are bacteria-caused destructions of the mineralised dental tissues,marked by the simultaneous activation of immune responses and regenerative events within the soft dental pulp tissue.While major molecular players in tooth decay have been uncovered during the past years,a detailed map of the molecular and cellular landscape of the diseased pulp is still missing.In this study we used single-cell RNA sequencing analysis,supplemented with immunostaining,to generate a comprehensive single-cell atlas of the pulp of carious human teeth.Our data demonstrated modifications in the various cell clusters within the pulp of carious teeth,such as immune cells,mesenchymal stem cells(MSC)and fibroblasts,when compared to the pulp of healthy human teeth.Active immune response in the carious pulp tissue is accompanied by specific changes in the fibroblast and MSC clusters.These changes include the upregulation of genes encoding extracellular matrix(ECM)components,including COL1A1 and Fibronectin(FN1),and the enrichment of the fibroblast cluster with myofibroblasts.The incremental changes in the ECM composition of carious pulp tissues were further confirmed by immunostaining analyses.Assessment of the Fibronectin fibres under mechanical strain conditions showed a significant tension reduction in carious pulp tissues,compared to the healthy ones.The present data demonstrate molecular,cellular and biomechanical alterations in the pulp of human carious teeth,indicative of extensive ECM remodelling,reminiscent of fibrosis observed in other organs.This comprehensive atlas of carious human teeth can facilitate future studies of dental pathologies and enable comparative analyses across diseased organs.展开更多
BACKGROUND Extracellular matrix(ECM)remodeling and stiffening,which are correlated with tumor malignancy,drives tumor development.However,the relationship between ECM remodeling and rat experimental model of 1,2-dimet...BACKGROUND Extracellular matrix(ECM)remodeling and stiffening,which are correlated with tumor malignancy,drives tumor development.However,the relationship between ECM remodeling and rat experimental model of 1,2-dimethylhyrazine(DMH)-induced colorectal cancer(CRC)imposed by cold and capsaicin exposure remains unclear.AIM To explore the effects of cold exposure and capsaicin on ECM remodeling and ECM enzymes in DMH-induced CRC.METHODS For histopathological analysis,the sections of colon tissues were stained with hematoxylin and eosin,Masson’s trichrome,Picrosirius red,and Weigert’s Resorcin-Fuchsin to observe the remodeling of collagen and elastin.Additionally,the protein expression level of type I collagen(COL I),type 3 collagen(COL III0,elastin,matrix metalloproteinase(MMP)1,MMP2,MMP9,and tissue-specific matrix metalloproteinase 1(TIMP1)was assessed by immunohistochemistry.The messenger RNA(mRNA)levels of COL I,COL III,elastin,and lysyl oxidase-like-2(LOXL2)in the colon tissues of rats was measured by reverse-transcriptase quantitative polymerase chain reaction.RESULTS Although no differences were observed in the proportion of adenomas,a trend towards the increase of invasive tumors was observed in the cold and capsaicin group.The cold exposure group had a metastasis rate compared with the other groups.Additionally,abnormal accumulation of both collagen and elastin was observed in the cold exposure and capsaicin group.Specifically,collagen quantitative analysis showed increased length,width,angle,and straightness compared with the DMH group.Collagen deposition and straightness were significantly increased in the cold exposure group compared with the capsaicin group.Cold exposure and capsaicin significantly increased the protein levels of COL I,elastin,and LOXL2 along with increases in their mRNA levels in the colon tissues compared with the DMH group,while COL III did not show a significant difference.Furthermore,in immunohistochemical evaluations,MMP1,MMP2,MMP9,and TIMP1 staining increased in the cold exposure and capsaicin group compared with the DMH group.CONCLUSION These results suggest that chronic cold and capsaicin exposure further increased the deposition of collagen and elastin in the colonic tissue.Increased COL I and elastin mRNA and protein levels expression may account for the enhanced ECM remodel and stiffness variations of colon tissue.The upregulated expression of the LOXL2 and physiological imbalance between MMP/TIMP activation and deactivation could contribute to the progression of the CRC resulting from cold and capsaicin exposure.展开更多
BACKGROUND The extracellular matrix is the main component of the tumor microenvironment.Extracellular matrix remodels with the oncogenesis and development of tumors.Previous studies usually focused on the changes of p...BACKGROUND The extracellular matrix is the main component of the tumor microenvironment.Extracellular matrix remodels with the oncogenesis and development of tumors.Previous studies usually focused on the changes of proteins in normal colorectal tissues and colorectal cancers.Little is known about the changes in the extracellular matrix in different stages of colorectal cancer and the effects of these changes on the development of this cancer.AIM To test the changes of type I collagen,type IV collagen,matrix metalloproteinase-2(MMP-2),matrix metalloproteinase-9(MMP-9),and tissue inhibitor of metalloproteinase-3(TIMP-3)in different stages of colorectal cancer and the effects of these changes on the proliferation of cancer cells.METHODS The extracellular matrix from various stages of colorectal cancer and normal colon tissue was obtained by using acellular technology.We used proteomics to detect the differential expression of proteins between normal colon tissues and colorectal cancer tissues,and then we used Western blot to observe their expression in each stage of colorectal cancer and in normal colon tissue.By coculturing the extracellular matrix and HT29 colon cancer cells in vivo and in vitro,we tested the cancer cell proliferation rate in vitro by methyl thiazolyl tetrazolium(MTT)assay and in vivo by measuring the tumor volume.RESULTS The expression of type I collagen and MMP-2 increased with increased tumor stage.The expression of MMP-9 was higher in colorectal cancer tissues and was highest in stage III cancer.The expression of type IV collagen and TIMP-3 decreased with increased tumor stage.The proliferation rate of cancer cells in the extracellular matrix of colorectal cancer was higher than that in the extracellular matrix of the normal colon.CONCLUSION These data suggest that the extracellular matrix structure and composition become disorganized during the development of tumors,which is more conducive for the growth of cancer cells.展开更多
Karacoline is a compound found in the plant Aconitum kusnezoffii Reichb.Although Aconitum kusnezoffii Reichb is widely used for the treatment of pain,very few studies have been carried out on the use of karacoline due...Karacoline is a compound found in the plant Aconitum kusnezoffii Reichb.Although Aconitum kusnezoffii Reichb is widely used for the treatment of pain,very few studies have been carried out on the use of karacoline due to its potential toxicity.In this study,we selected key matrix metalloproteinases(MMPs),collagen II,and aggrecan as targets due to their association with intervertebral disc degeneration(IDD).Using these targets,we then used network pharmacology to predict a series of molecules that might exert therapeutic effects on IDD.Of these molecules,karacoline was predicted to have the best effect.Tumor necrosis factor(TNF)-a is known to promote the degeneration of the extracellular matrix in IDD.We therefore applied different concentrations of karacoline(0,1.25,or 12.88 mM)along with 100 ng/mL TNF-a to rat nucleus pulposus cells and found that karacoline reduced the expression of MMP-14 in IDD by inhibiting the nuclear factor(NF)-κB pathway,while collagen II and aggrecan expression was increased.This suggested that extracellular matrix degradation was inhibited by karacoline(P<0.05).Our data therefore reveal a new clinical application of karacoline and provide support for the use of network pharmacology in predicting novel drugs.展开更多
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 ge...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.展开更多
Extracellular matrix( ECM) plays a prominent role in establishing and maintaining an appropriate microenvironment for tissue regeneration. The aims of this study were to construct a tissue engineered scaffold by recon...Extracellular matrix( ECM) plays a prominent role in establishing and maintaining an appropriate microenvironment for tissue regeneration. The aims of this study were to construct a tissue engineered scaffold by reconstituting osteoblast cell-derived ECM( O-ECM) on the electrospun nanofibrous scaffold,and further to evaluate its subsequent application for promoting the proliferation of bone marrow mesenchymal stem cells( BMSCs). To engineer a biomimetic scaffold, calvarial osteoblasts and electrospun poly-llactic acid( PLLA) nanofibers were prepared and subjected to decellularize for O-ECM deposition. To evaluate and characterize the O-ECM/PLLA scaffold, the morphology was examined and several specific mark proteins of osteoblasts matrix were evaluated.Furthermore,the cell counting kit-8( CCK-8) assay was used to detect the proliferation of the BMSCs cultivated on the O-ECM/PLLA scaffold. The results indicated O-ECM/PLLA scaffold was loaded with Collagen I, Fibronectin, and Laminin, as the composition of the marrow ECM. After decellularization,O-ECM deposition was observed in O-ECM/PLLA scaffold. Moreover,the O-ECM/PLLA scaffold could significantly enhance the proliferation of BMSCs,suggesting better cytocompatibility compared to the other groups tested. Taken together,a biomimetic scaffold based on the joint use of O-ECM and PLLA biomaterials,which represents a promising approach to bone tissue engineering, facilitates the expansion of BMSCs in vitro.展开更多
The extracellular matrix(ECM)comprises of many structural molecules that constitute the extracellular environment.ECM molecules are characterized by specific features like diversity,complexity and signaling,which are ...The extracellular matrix(ECM)comprises of many structural molecules that constitute the extracellular environment.ECM molecules are characterized by specific features like diversity,complexity and signaling,which are also results of improvement or development of disease mediated by some physiological changes.Several drugs have also been used to manage diseases and they have been reported to modulate ECM assembly,including physiological changes,beyond their primary targets and ECM metabolism.This review highlights the alteration of ECM environment for diseases and effect of different classes of drugs like nonsteroidal anti-inflammatory drugs,immune suppressant drug,steroids on ECM or its components.Thus,it is summarized from previously conducted researches that diseases can be managed by targeting specific components of ECM which are involved in the pathophysiology of diseases.Moreover,the drug delivery focused on targeting the ECM components also has the potential for the discovery of targeted and site specific release of drugs.Therefore,ECM or its components could be future targets for the development of new drugs for controlling various disease conditions including neurodegenerative diseases and cancers.展开更多
AIM: To investigate changes in extracellular matrix(ECM) gene expression in human trabecular meshwork(HTM) cells in response to mechanical fluid flow stimulation.METHODS: HTM cells were grown on a glass plate coated w...AIM: To investigate changes in extracellular matrix(ECM) gene expression in human trabecular meshwork(HTM) cells in response to mechanical fluid flow stimulation.METHODS: HTM cells were grown on a glass plate coated with 0.02% type Ⅰ collagen(COL) and exposed to shear stress(0, 0.2, 1.0 dyne/cm;) for 12 h.Changes in genes related to the ECM were evaluated by real-time reverse transcriptase-polymerase chain reaction.Phosphorylation of Smad2 protein was investigated by Western blotting.RESULTS: After mechanical stimulation, COL type 4 alpha 2, COL type 6 alpha 1, and fibronectin-1 mRNA were significantly higher than the static control(P<0.05, <0.05, and <0.01, respectively).The metalloproteinase-2 and plasminogen activator inhibitor-1 mRNA were significantly higher than the static control(P<0.05 and <0.01, respectively), while the differences in the tissue inhibitors of metalloproteinases-2 mRNA were not significant.The phosphorylation of Smad2 levels was significantly higher compared to the static control cells.CONCLUSION: Changes in the expressions of genes associated ECM metabolism result in HTM cells after mechanical stimulation.The mechanical stimulation of the aqueous humor to the trabecular meshwork may promote ECM turnover and contribute to intraocular pressure homeostasis.展开更多
In the glomeruli,mesangial cells produce mesangial matrix while podocytes wrap glomerular capillaries with cellular extensions named foot processes and tether the glomerular basement membrane(GBM).The turnover of the ...In the glomeruli,mesangial cells produce mesangial matrix while podocytes wrap glomerular capillaries with cellular extensions named foot processes and tether the glomerular basement membrane(GBM).The turnover of the mature GBM and the ability of adult podocytes to repair injured GBM are unclear.The actin cytoskeleton is a major cytoplasmic component of podocyte foot processes and links the cell to the GBM.Predominant components of the normal glomerular extracellular matrix(ECM)include glycosaminoglycans,proteoglycans,laminins,fibronectin-1,and several types of collagen.In patients with diabetes,multiorgan composition of extracellular tissues is anomalous,including the kidney,so that the constitution and arrangement of glomerular ECM is profoundly altered.In patients with diabetic kidney disease(DKD),the global quantity of glomerular ECM is increased.The level of sulfated proteoglycans is reduced while hyaluronic acid is augmented,compared to control subjects.The concentration of mesangial fibronectin-1 varies depending on the stage of DKD.Mesangial type III collagen is abundant in patients with DKD,unlike normal kidneys.The amount of type V and type VI collagens is higher in DKD and increases with the progression of the disease.The GBM contains lower amount of type IV collagen in DKD compared to normal tissue.Further,genetic variants in theα3 chain of type IV collagen may modulate susceptibility to DKD and end-stage kidney disease.Human cellular models of glomerular cells,analyses of human glomerular proteome,and improved microscopy procedures have been developed to investigate the molecular composition and organization of the human glomerular ECM.展开更多
The importance of the extracellular matrix(ECM) in contributing to structural,mechanical,functional and tissue-specific features in the body is well appreciated. While the ECM was previously considered to be a passive...The importance of the extracellular matrix(ECM) in contributing to structural,mechanical,functional and tissue-specific features in the body is well appreciated. While the ECM was previously considered to be a passive bystander,it is now evident that it plays active,dynamic and flexible roles in shaping cell survival,differentiation,migration and death to varying extents depending on the specific site in the body. Dendritic cells(DCs) are recognized as potent antigen presenting cells present in many tissues and in blood,continuously scrutinizing the microenvironment for antigens and mounting local and systemic host responses against harmful agents. DCs also play pivotal roles in maintaining homeostasis to harmless self-antigens,critical for preventing autoimmunity. What is less understood are the complex interactions between DCs and the ECM in maintaining this balance between steady-state tissue residence and DC activation during inflammation. DCs are finely tuned to inflammationinduced variations in fragment length,accessible epitopes and post-translational modifications of individual ECM components and correspondingly interpret these changes appropriately by adjusting their profiles of cognate binding receptors and downstream immune activation. The successful design and composition of novel ECMbased mimetics in regenerative medicine and other applications rely on our improved understanding of DCECM interplay in homeostasis and the challenges involved in maintaining it.展开更多
Regeneration in the central nervous system(CNS)is limited,and CNS damage often leads to cognitive impairment or permanent functional motor and sensory loss.Impaired regenerative capacity is multifactorial and includes...Regeneration in the central nervous system(CNS)is limited,and CNS damage often leads to cognitive impairment or permanent functional motor and sensory loss.Impaired regenerative capacity is multifactorial and includes inflammation,loss of the bloodbrain barrier,and alteration in the extracellular matrix(ECM).One of the main problems is the formation of a glial scar and the production of inhibitory ECM,such as proteoglycans。展开更多
Estrogen is essential for the skin to maintain its physiological function. The binding of estrogen to the estrogen receptor (ER) activates gene transcription, which has biological effects on the target tissue. Estroge...Estrogen is essential for the skin to maintain its physiological function. The binding of estrogen to the estrogen receptor (ER) activates gene transcription, which has biological effects on the target tissue. Estrogen levels and ER expression are known to decrease with aging and exposure to ultraviolet light (UV);therefore, increased estrogen levels and ER expression may improve age-related changes in the skin. <em>Rehmannia</em> root has been reported to have blood circulation-promoting and anti-inflammatory effects;however, few studies have reported the effects of <em>Rehmannia</em> root on skin. In this study, we examined the effects of <em>Rehmannia glutinosa</em> Libosch. var. purpurea Makino root extract (RE) on ER expression, and estrogen, RE, or their related ingredients increased ER expression in human epidermal keratinocytes, human dermal fibroblasts, and skin models. Moreover, RE increased the production of basic fibroblast growth factor, transforming growth factor <em>β</em>1, and epidermal growth factor. The mixture of estrogen and RE improved extracellular matrix (ECM) production to a greater degree than estrogen and RE independently. Although high population doubling levels (PDL) and UV irradiation downregulated ER expression, RE upregulated ER expression in high PDL cells and UV irradiated cells. In addition, RE increased the expression of epidermal differentiation marker proteins compared to their expression levels in the absence of RE. The collective findings suggest that RE aids in the prevention of skin aging by upregulating the ER expression that has been decreased by aging and UV and promoting estrogen activity, ECM production, and epidermal differentiation.展开更多
AIM: To investigate a new technique of the anorectal fistula treatment with acellular extracellular matrix (AEM). METHODS: Thirty patients with anorectal fistula were treated with AEM. All fistula tracts and primary o...AIM: To investigate a new technique of the anorectal fistula treatment with acellular extracellular matrix (AEM). METHODS: Thirty patients with anorectal fistula were treated with AEM. All fistula tracts and primary openings were identified using conventional fistula probe. All tracts were curetted with curet and irrigated with hydrogen peroxide and metronidazole. The AEM was pulled into the fistula tract from secondary to primary opening. The material was secured at the level of the primary opening. The excess AEM was trimmed at skin level at the secondary opening. RESULTS: All of the 30 patients had successful closure of their fistula after a 7-14 d follow-up. The healing rate of anal fistula in treatment group was 100%. The ache time, healing time and anal deformation of treatment group were obviously superior to traditional surgical methods. CONCLUSION: Using AEM anal fistula plug in treatment that causes the anorectal fistula is safe and successful in 100% of patients. It can reduce pain, shorten disease course and protect anal function.展开更多
Background: Endothelial and smooth muscle cells were used as seeding cells and heterogeneous acellularized matrix was used as scaffold to construct the tissue-engineered graft. Methods: A 2 weeks piglet was selected a...Background: Endothelial and smooth muscle cells were used as seeding cells and heterogeneous acellularized matrix was used as scaffold to construct the tissue-engineered graft. Methods: A 2 weeks piglet was selected as a donor of seeding cells. Two-centimetre length of common carotid artery was dissected. Endothelial cells and smooth muscle cells were harvested by trypsin and collagenase digestion respectively. The isolated cells were cultured and expanded using routine cell culture technique. An adult sheep was used as a donor of acellularized matrix. The thoracic aorta was harvested and processed by a multi-step de-cellularizing technique to remove the original cells and preserve the elastic and collagen fibers. The cultured smooth muscle cells and endothelial cells were then seeded to the acellularized matrix and incubated in vitro for another 2 weeks. The cell seeded graft was then transplanted to the cell-donated piglet to substitute part of the native pulmonary artery. Results: The cultured cells from piglet were characterized as endothelial cells by the presence of specific antigens vWF and CD31, and smooth muscle cells by the presence of specific antigen α-actin on the cell surface respectively with immunohistochemical technique. After decellularizing processing for the thoracic aorta from sheep, all the cellular components were extracted and elastic and collagen fibers kept their original morphology and structure. The maximal load of acellular matrix was decreased and 20% lower than that of untreated thoracic aorta, but the maximal tensions between them were not different statistically and they had similar load-tension curves. Three months after transplantation, the animal was sacrificed and the graft was removed for observation. The results showed that the inner surfaces of the graft were smooth, without thrombosis and calcification. Under microscopy, a great number of growing cells could be seen and elastic and collagen fibers were abundant. Conclusion: Cultured self-derived endothelial and smooth muscle cells could be used as seeding cells and heterogeneous acellularized matrix could be used as scaffold in constructing tissue-engineered graft.展开更多
Arterial embolization of myomas (AEM) is an established option for the conservative treatment of uterine leio-myomas;it treats all present uterine nodules at once, is less invasive than other procedures and effective ...Arterial embolization of myomas (AEM) is an established option for the conservative treatment of uterine leio-myomas;it treats all present uterine nodules at once, is less invasive than other procedures and effective in controlling symptoms, and does not require long term hospitalizations. Nevertheless, the potential impact on endometrial morphological and functional outcomes after the procedure is still controversial based on reports of endometritis or eventual transient ischemia. This study evaluated endometrial reorganization in uterine leiomyoma patients, before and after AEM, through gene expression analyses of extracellular matrix and cytokines genes in theendometrial tissue. Eight patients with leiomyomas were evaluated before AEM and 6 months after. The examinations included transvaginal pelvic ultrasonography, dosing of the follicle-stimulating hormone, and endometrial biopsy during the second phase of the menstrual cycle. RNA was extracted from endometrial samples, cDNA was synthesized, and applied on PCR arrayTM plates to evaluate the expression of extracellular matrix (ECM) genes and cytokines and their receptors’ genes (CYT). The ECM overexpressed genes were MMP (1, 3, 10, 11, and 14), CTGF1, ICAM1, TBHS1, ITGA2, ITGA3, ITGB3, COL7A1, COL12A, SPP1, and TNC;ADAMTS8 was underexpressed. The CYT overexpressed genes were SPP1, BCL6, CXCL12, IL-8, and CEBPB;CXCL13 and CCL21 were underexpressed. The ECM results showed overexpression of proteases that are responsible for dysfunctions in the ECM, and of genes responsible for adhesion and membrane components. The CYT results showed overexpression of chemokines responsible for endometrial repair, and underexpression of cytokines involved in inflammatory processes in the endometrial tissue. AEM treatment did not negatively affect the endometrial function at 6 months after embolization. This study broadens the knowledge about using a procedure that is relevant to the treatment of leiomyomas and contributes to the establishment of future guidelines for the decision making process for physicians and patients.展开更多
Song et al have reported a 100% success rate of acellular extracellular matrix (AEM) anal fistula plug in low fistula-in-ano. The results with this product in high fistula-in-ano are keenly awaited.
Objective To establish a method for quantitative detection of the sulfate glycosaminoglycans ( GAG) content in extracellular matrix of in vitro cultured chondrocytes so as to evaluate the biological characteristics of...Objective To establish a method for quantitative detection of the sulfate glycosaminoglycans ( GAG) content in extracellular matrix of in vitro cultured chondrocytes so as to evaluate the biological characteristics of epiphyseal, articular and rib chondrocytes. Methods Sulfate GAG content in extracellular matrix of three chondrocytes was measured by the modified dimethylmethylene blue (DMB) method. The changes of the toluidine blue (TB) stain of chondrocytes were observed by light microscope. Results Primary chondrocytes had the highest content of sulfate GAG in the extracellular matrix, ie, epiphyseal chondrocytes reached ( 70. 12 ± 7. 72 )μg/cm2, articular chondrocytes (92.00 ± 10.15) μg/cm2 and rib chondrocytes (80.61 ± 11. 40) μg/cm2, respectively. On the third pasage chondrocytes, epiphyceal chondrocytes decreased to (53.27 ± 9. 50 ) μg/cm2, articular chondrocytes to (63.88 ± 11.92) μg/cm2 and rib chondrocytes to (58.94 ±8.21) μg/cm2, respectively. The change of TB in every passage展开更多
基金National Natural Science Foundation of China,No.U20A20403This study was conducted in accordance with the Animal Ethics Committee of the Institute of Antler Science and Product Technology,Changchun Sci-Tech University(AEC No:CKARI202309).
文摘BACKGROUND Cartilage defects are some of the most common causes of arthritis.Cartilage lesions caused by inflammation,trauma or degenerative disease normally result in osteochondral defects.Previous studies have shown that decellularized extracellular matrix(ECM)derived from autologous,allogenic,or xenogeneic mesenchymal stromal cells(MSCs)can effectively restore osteochondral integrity.AIM To determine whether the decellularized ECM of antler reserve mesenchymal cells(RMCs),a xenogeneic material from antler stem cells,is superior to the currently available treatments for osteochondral defects.METHODS We isolated the RMCs from a 60-d-old sika deer antler and cultured them in vitro to 70%confluence;50 mg/mL L-ascorbic acid was then added to the medium to stimulate ECM deposition.Decellularized sheets of adipocyte-derived MSCs(aMSCs)and antlerogenic periosteal cells(another type of antler stem cells)were used as the controls.Three weeks after ascorbic acid stimulation,the ECM sheets were harvested and applied to the osteochondral defects in rat knee joints.RESULTS The defects were successfully repaired by applying the ECM-sheets.The highest quality of repair was achieved in the RMC-ECM group both in vitro(including cell attachment and proliferation),and in vivo(including the simultaneous regeneration of well-vascularized subchondral bone and avascular articular hyaline cartilage integrated with surrounding native tissues).Notably,the antler-stem-cell-derived ECM(xenogeneic)performed better than the aMSC-ECM(allogenic),while the ECM of the active antler stem cells was superior to that of the quiescent antler stem cells.CONCLUSION Decellularized xenogeneic ECM derived from the antler stem cell,particularly the active form(RMC-ECM),can achieve high quality repair/reconstruction of osteochondral defects,suggesting that selection of decellularized ECM for such repair should be focused more on bioactivity rather than kinship.
基金Supported by the Key Program of the Natural Science Foundation of Ningbo,No.202003N4019Ningbo City COVID-19 Epidemic Prevention and Control Project,No.202002N7033+1 种基金Zhejiang Provincial Natural Science Foundation of China,No.Y23H190011Zhejiang Medical and Health Science and Technology Project,No.2019KY154。
文摘An outbreak of coronavirus disease 2019(COVID-19)has spread globally,with over 500 million cases and 6 million deaths to date.COVID-19 is associated with a systemic inflammatory response and abnormalities of the extracellular matrix(ECM),which is also involved in inflammatory storms.Upon viral infection,ECM proteins are involved in the recruitment of inflammatory cells and interference with target organ metabolism,including in the lungs.Additionally,serum biomarkers of ECM turnover are associated with the severity of COVID-19 and may serve as potential targets.Consequently,understanding the expression and function of ECM,particularly of the lung,during severe acute respiratory syndrome of the coronavirus 2 infection would provide valuable insights into the mechanisms of COVID-19 progression.In this review,we summarize the current findings on ECM,such as hyaluronic acid,matrix metalloproteinases,and collagen,which are linked to the severity and inflammation of COVID-19.Some drugs targeting the extracellular surface have been effective.In the future,these ECM findings could provide novel perspectives on the pathogenesis and treatment of COVID-19.
基金supported by continuation funds from the Turku Collegium for Science,Medicine and Technologythe Japan Society for the Promotion of Science (#23K08670)+1 种基金the Sigrid Jusélius Foundation (#230131)MF-R internship at the University of Turku was funded by the Erasmus+program。
文摘The bone extracellular matrix(ECM) contains minerals deposited on highly crosslinked collagen fibrils and hundreds of noncollagenous proteins. Some of these proteins are key to the regulation of bone formation and regeneration via signaling pathways,and play important regulatory and structural roles. However, the complete list of bone extracellular matrix proteins, their roles, and the extent of individual and cross-species variations have not been fully captured in both humans and model organisms. Here, we introduce the most comprehensive resource of bone extracellular matrix(ECM) proteins that can be used in research fields such as bone regeneration, osteoporosis, and mechanobiology. The Phylobone database(available at https://phylobone.com) includes 255proteins potentially expressed in the bone extracellular matrix(ECM) of humans and 30 species of vertebrates. A bioinformatics pipeline was used to identify the evolutionary relationships of bone ECM proteins. The analysis facilitated the identification of potential model organisms to study the molecular mechanisms of bone regeneration. A network analysis showed high connectivity of bone ECM proteins. A total of 214 functional protein domains were identified, including collagen and the domains involved in bone formation and resorption. Information from public drug repositories was used to identify potential repurposing of existing drugs. The Phylobone database provides a platform to study bone regeneration and osteoporosis in light of(biological) evolution,and will substantially contribute to the identification of molecular mechanisms and drug targets.
文摘Carious lesions are bacteria-caused destructions of the mineralised dental tissues,marked by the simultaneous activation of immune responses and regenerative events within the soft dental pulp tissue.While major molecular players in tooth decay have been uncovered during the past years,a detailed map of the molecular and cellular landscape of the diseased pulp is still missing.In this study we used single-cell RNA sequencing analysis,supplemented with immunostaining,to generate a comprehensive single-cell atlas of the pulp of carious human teeth.Our data demonstrated modifications in the various cell clusters within the pulp of carious teeth,such as immune cells,mesenchymal stem cells(MSC)and fibroblasts,when compared to the pulp of healthy human teeth.Active immune response in the carious pulp tissue is accompanied by specific changes in the fibroblast and MSC clusters.These changes include the upregulation of genes encoding extracellular matrix(ECM)components,including COL1A1 and Fibronectin(FN1),and the enrichment of the fibroblast cluster with myofibroblasts.The incremental changes in the ECM composition of carious pulp tissues were further confirmed by immunostaining analyses.Assessment of the Fibronectin fibres under mechanical strain conditions showed a significant tension reduction in carious pulp tissues,compared to the healthy ones.The present data demonstrate molecular,cellular and biomechanical alterations in the pulp of human carious teeth,indicative of extensive ECM remodelling,reminiscent of fibrosis observed in other organs.This comprehensive atlas of carious human teeth can facilitate future studies of dental pathologies and enable comparative analyses across diseased organs.
基金by National Natural Science Foundation of China,No.81673944.
文摘BACKGROUND Extracellular matrix(ECM)remodeling and stiffening,which are correlated with tumor malignancy,drives tumor development.However,the relationship between ECM remodeling and rat experimental model of 1,2-dimethylhyrazine(DMH)-induced colorectal cancer(CRC)imposed by cold and capsaicin exposure remains unclear.AIM To explore the effects of cold exposure and capsaicin on ECM remodeling and ECM enzymes in DMH-induced CRC.METHODS For histopathological analysis,the sections of colon tissues were stained with hematoxylin and eosin,Masson’s trichrome,Picrosirius red,and Weigert’s Resorcin-Fuchsin to observe the remodeling of collagen and elastin.Additionally,the protein expression level of type I collagen(COL I),type 3 collagen(COL III0,elastin,matrix metalloproteinase(MMP)1,MMP2,MMP9,and tissue-specific matrix metalloproteinase 1(TIMP1)was assessed by immunohistochemistry.The messenger RNA(mRNA)levels of COL I,COL III,elastin,and lysyl oxidase-like-2(LOXL2)in the colon tissues of rats was measured by reverse-transcriptase quantitative polymerase chain reaction.RESULTS Although no differences were observed in the proportion of adenomas,a trend towards the increase of invasive tumors was observed in the cold and capsaicin group.The cold exposure group had a metastasis rate compared with the other groups.Additionally,abnormal accumulation of both collagen and elastin was observed in the cold exposure and capsaicin group.Specifically,collagen quantitative analysis showed increased length,width,angle,and straightness compared with the DMH group.Collagen deposition and straightness were significantly increased in the cold exposure group compared with the capsaicin group.Cold exposure and capsaicin significantly increased the protein levels of COL I,elastin,and LOXL2 along with increases in their mRNA levels in the colon tissues compared with the DMH group,while COL III did not show a significant difference.Furthermore,in immunohistochemical evaluations,MMP1,MMP2,MMP9,and TIMP1 staining increased in the cold exposure and capsaicin group compared with the DMH group.CONCLUSION These results suggest that chronic cold and capsaicin exposure further increased the deposition of collagen and elastin in the colonic tissue.Increased COL I and elastin mRNA and protein levels expression may account for the enhanced ECM remodel and stiffness variations of colon tissue.The upregulated expression of the LOXL2 and physiological imbalance between MMP/TIMP activation and deactivation could contribute to the progression of the CRC resulting from cold and capsaicin exposure.
文摘BACKGROUND The extracellular matrix is the main component of the tumor microenvironment.Extracellular matrix remodels with the oncogenesis and development of tumors.Previous studies usually focused on the changes of proteins in normal colorectal tissues and colorectal cancers.Little is known about the changes in the extracellular matrix in different stages of colorectal cancer and the effects of these changes on the development of this cancer.AIM To test the changes of type I collagen,type IV collagen,matrix metalloproteinase-2(MMP-2),matrix metalloproteinase-9(MMP-9),and tissue inhibitor of metalloproteinase-3(TIMP-3)in different stages of colorectal cancer and the effects of these changes on the proliferation of cancer cells.METHODS The extracellular matrix from various stages of colorectal cancer and normal colon tissue was obtained by using acellular technology.We used proteomics to detect the differential expression of proteins between normal colon tissues and colorectal cancer tissues,and then we used Western blot to observe their expression in each stage of colorectal cancer and in normal colon tissue.By coculturing the extracellular matrix and HT29 colon cancer cells in vivo and in vitro,we tested the cancer cell proliferation rate in vitro by methyl thiazolyl tetrazolium(MTT)assay and in vivo by measuring the tumor volume.RESULTS The expression of type I collagen and MMP-2 increased with increased tumor stage.The expression of MMP-9 was higher in colorectal cancer tissues and was highest in stage III cancer.The expression of type IV collagen and TIMP-3 decreased with increased tumor stage.The proliferation rate of cancer cells in the extracellular matrix of colorectal cancer was higher than that in the extracellular matrix of the normal colon.CONCLUSION These data suggest that the extracellular matrix structure and composition become disorganized during the development of tumors,which is more conducive for the growth of cancer cells.
文摘Karacoline is a compound found in the plant Aconitum kusnezoffii Reichb.Although Aconitum kusnezoffii Reichb is widely used for the treatment of pain,very few studies have been carried out on the use of karacoline due to its potential toxicity.In this study,we selected key matrix metalloproteinases(MMPs),collagen II,and aggrecan as targets due to their association with intervertebral disc degeneration(IDD).Using these targets,we then used network pharmacology to predict a series of molecules that might exert therapeutic effects on IDD.Of these molecules,karacoline was predicted to have the best effect.Tumor necrosis factor(TNF)-a is known to promote the degeneration of the extracellular matrix in IDD.We therefore applied different concentrations of karacoline(0,1.25,or 12.88 mM)along with 100 ng/mL TNF-a to rat nucleus pulposus cells and found that karacoline reduced the expression of MMP-14 in IDD by inhibiting the nuclear factor(NF)-κB pathway,while collagen II and aggrecan expression was increased.This suggested that extracellular matrix degradation was inhibited by karacoline(P<0.05).Our data therefore reveal a new clinical application of karacoline and provide support for the use of network pharmacology in predicting novel drugs.
文摘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.
基金Shanghai Municipal Natural Science Foundation,China(No.15ZR1400500)the Fundamental Research Funds for the Central Universities,China(Nos.16D110520,EG2017011)
文摘Extracellular matrix( ECM) plays a prominent role in establishing and maintaining an appropriate microenvironment for tissue regeneration. The aims of this study were to construct a tissue engineered scaffold by reconstituting osteoblast cell-derived ECM( O-ECM) on the electrospun nanofibrous scaffold,and further to evaluate its subsequent application for promoting the proliferation of bone marrow mesenchymal stem cells( BMSCs). To engineer a biomimetic scaffold, calvarial osteoblasts and electrospun poly-llactic acid( PLLA) nanofibers were prepared and subjected to decellularize for O-ECM deposition. To evaluate and characterize the O-ECM/PLLA scaffold, the morphology was examined and several specific mark proteins of osteoblasts matrix were evaluated.Furthermore,the cell counting kit-8( CCK-8) assay was used to detect the proliferation of the BMSCs cultivated on the O-ECM/PLLA scaffold. The results indicated O-ECM/PLLA scaffold was loaded with Collagen I, Fibronectin, and Laminin, as the composition of the marrow ECM. After decellularization,O-ECM deposition was observed in O-ECM/PLLA scaffold. Moreover,the O-ECM/PLLA scaffold could significantly enhance the proliferation of BMSCs,suggesting better cytocompatibility compared to the other groups tested. Taken together,a biomimetic scaffold based on the joint use of O-ECM and PLLA biomaterials,which represents a promising approach to bone tissue engineering, facilitates the expansion of BMSCs in vitro.
文摘The extracellular matrix(ECM)comprises of many structural molecules that constitute the extracellular environment.ECM molecules are characterized by specific features like diversity,complexity and signaling,which are also results of improvement or development of disease mediated by some physiological changes.Several drugs have also been used to manage diseases and they have been reported to modulate ECM assembly,including physiological changes,beyond their primary targets and ECM metabolism.This review highlights the alteration of ECM environment for diseases and effect of different classes of drugs like nonsteroidal anti-inflammatory drugs,immune suppressant drug,steroids on ECM or its components.Thus,it is summarized from previously conducted researches that diseases can be managed by targeting specific components of ECM which are involved in the pathophysiology of diseases.Moreover,the drug delivery focused on targeting the ECM components also has the potential for the discovery of targeted and site specific release of drugs.Therefore,ECM or its components could be future targets for the development of new drugs for controlling various disease conditions including neurodegenerative diseases and cancers.
基金Supported by JSPS KAKENHI (No.15K10857No.17H00695No.19K18865)。
文摘AIM: To investigate changes in extracellular matrix(ECM) gene expression in human trabecular meshwork(HTM) cells in response to mechanical fluid flow stimulation.METHODS: HTM cells were grown on a glass plate coated with 0.02% type Ⅰ collagen(COL) and exposed to shear stress(0, 0.2, 1.0 dyne/cm;) for 12 h.Changes in genes related to the ECM were evaluated by real-time reverse transcriptase-polymerase chain reaction.Phosphorylation of Smad2 protein was investigated by Western blotting.RESULTS: After mechanical stimulation, COL type 4 alpha 2, COL type 6 alpha 1, and fibronectin-1 mRNA were significantly higher than the static control(P<0.05, <0.05, and <0.01, respectively).The metalloproteinase-2 and plasminogen activator inhibitor-1 mRNA were significantly higher than the static control(P<0.05 and <0.01, respectively), while the differences in the tissue inhibitors of metalloproteinases-2 mRNA were not significant.The phosphorylation of Smad2 levels was significantly higher compared to the static control cells.CONCLUSION: Changes in the expressions of genes associated ECM metabolism result in HTM cells after mechanical stimulation.The mechanical stimulation of the aqueous humor to the trabecular meshwork may promote ECM turnover and contribute to intraocular pressure homeostasis.
文摘In the glomeruli,mesangial cells produce mesangial matrix while podocytes wrap glomerular capillaries with cellular extensions named foot processes and tether the glomerular basement membrane(GBM).The turnover of the mature GBM and the ability of adult podocytes to repair injured GBM are unclear.The actin cytoskeleton is a major cytoplasmic component of podocyte foot processes and links the cell to the GBM.Predominant components of the normal glomerular extracellular matrix(ECM)include glycosaminoglycans,proteoglycans,laminins,fibronectin-1,and several types of collagen.In patients with diabetes,multiorgan composition of extracellular tissues is anomalous,including the kidney,so that the constitution and arrangement of glomerular ECM is profoundly altered.In patients with diabetic kidney disease(DKD),the global quantity of glomerular ECM is increased.The level of sulfated proteoglycans is reduced while hyaluronic acid is augmented,compared to control subjects.The concentration of mesangial fibronectin-1 varies depending on the stage of DKD.Mesangial type III collagen is abundant in patients with DKD,unlike normal kidneys.The amount of type V and type VI collagens is higher in DKD and increases with the progression of the disease.The GBM contains lower amount of type IV collagen in DKD compared to normal tissue.Further,genetic variants in theα3 chain of type IV collagen may modulate susceptibility to DKD and end-stage kidney disease.Human cellular models of glomerular cells,analyses of human glomerular proteome,and improved microscopy procedures have been developed to investigate the molecular composition and organization of the human glomerular ECM.
基金Supported by The Royal College of Surgeons of Edinburgh
文摘The importance of the extracellular matrix(ECM) in contributing to structural,mechanical,functional and tissue-specific features in the body is well appreciated. While the ECM was previously considered to be a passive bystander,it is now evident that it plays active,dynamic and flexible roles in shaping cell survival,differentiation,migration and death to varying extents depending on the specific site in the body. Dendritic cells(DCs) are recognized as potent antigen presenting cells present in many tissues and in blood,continuously scrutinizing the microenvironment for antigens and mounting local and systemic host responses against harmful agents. DCs also play pivotal roles in maintaining homeostasis to harmless self-antigens,critical for preventing autoimmunity. What is less understood are the complex interactions between DCs and the ECM in maintaining this balance between steady-state tissue residence and DC activation during inflammation. DCs are finely tuned to inflammationinduced variations in fragment length,accessible epitopes and post-translational modifications of individual ECM components and correspondingly interpret these changes appropriately by adjusting their profiles of cognate binding receptors and downstream immune activation. The successful design and composition of novel ECMbased mimetics in regenerative medicine and other applications rely on our improved understanding of DCECM interplay in homeostasis and the challenges involved in maintaining it.
文摘Regeneration in the central nervous system(CNS)is limited,and CNS damage often leads to cognitive impairment or permanent functional motor and sensory loss.Impaired regenerative capacity is multifactorial and includes inflammation,loss of the bloodbrain barrier,and alteration in the extracellular matrix(ECM).One of the main problems is the formation of a glial scar and the production of inhibitory ECM,such as proteoglycans。
文摘Estrogen is essential for the skin to maintain its physiological function. The binding of estrogen to the estrogen receptor (ER) activates gene transcription, which has biological effects on the target tissue. Estrogen levels and ER expression are known to decrease with aging and exposure to ultraviolet light (UV);therefore, increased estrogen levels and ER expression may improve age-related changes in the skin. <em>Rehmannia</em> root has been reported to have blood circulation-promoting and anti-inflammatory effects;however, few studies have reported the effects of <em>Rehmannia</em> root on skin. In this study, we examined the effects of <em>Rehmannia glutinosa</em> Libosch. var. purpurea Makino root extract (RE) on ER expression, and estrogen, RE, or their related ingredients increased ER expression in human epidermal keratinocytes, human dermal fibroblasts, and skin models. Moreover, RE increased the production of basic fibroblast growth factor, transforming growth factor <em>β</em>1, and epidermal growth factor. The mixture of estrogen and RE improved extracellular matrix (ECM) production to a greater degree than estrogen and RE independently. Although high population doubling levels (PDL) and UV irradiation downregulated ER expression, RE upregulated ER expression in high PDL cells and UV irradiated cells. In addition, RE increased the expression of epidermal differentiation marker proteins compared to their expression levels in the absence of RE. The collective findings suggest that RE aids in the prevention of skin aging by upregulating the ER expression that has been decreased by aging and UV and promoting estrogen activity, ECM production, and epidermal differentiation.
文摘AIM: To investigate a new technique of the anorectal fistula treatment with acellular extracellular matrix (AEM). METHODS: Thirty patients with anorectal fistula were treated with AEM. All fistula tracts and primary openings were identified using conventional fistula probe. All tracts were curetted with curet and irrigated with hydrogen peroxide and metronidazole. The AEM was pulled into the fistula tract from secondary to primary opening. The material was secured at the level of the primary opening. The excess AEM was trimmed at skin level at the secondary opening. RESULTS: All of the 30 patients had successful closure of their fistula after a 7-14 d follow-up. The healing rate of anal fistula in treatment group was 100%. The ache time, healing time and anal deformation of treatment group were obviously superior to traditional surgical methods. CONCLUSION: Using AEM anal fistula plug in treatment that causes the anorectal fistula is safe and successful in 100% of patients. It can reduce pain, shorten disease course and protect anal function.
基金Project (No. 99ZB14018) supported by the Natural Science Foun-dation of Shanghai, China
文摘Background: Endothelial and smooth muscle cells were used as seeding cells and heterogeneous acellularized matrix was used as scaffold to construct the tissue-engineered graft. Methods: A 2 weeks piglet was selected as a donor of seeding cells. Two-centimetre length of common carotid artery was dissected. Endothelial cells and smooth muscle cells were harvested by trypsin and collagenase digestion respectively. The isolated cells were cultured and expanded using routine cell culture technique. An adult sheep was used as a donor of acellularized matrix. The thoracic aorta was harvested and processed by a multi-step de-cellularizing technique to remove the original cells and preserve the elastic and collagen fibers. The cultured smooth muscle cells and endothelial cells were then seeded to the acellularized matrix and incubated in vitro for another 2 weeks. The cell seeded graft was then transplanted to the cell-donated piglet to substitute part of the native pulmonary artery. Results: The cultured cells from piglet were characterized as endothelial cells by the presence of specific antigens vWF and CD31, and smooth muscle cells by the presence of specific antigen α-actin on the cell surface respectively with immunohistochemical technique. After decellularizing processing for the thoracic aorta from sheep, all the cellular components were extracted and elastic and collagen fibers kept their original morphology and structure. The maximal load of acellular matrix was decreased and 20% lower than that of untreated thoracic aorta, but the maximal tensions between them were not different statistically and they had similar load-tension curves. Three months after transplantation, the animal was sacrificed and the graft was removed for observation. The results showed that the inner surfaces of the graft were smooth, without thrombosis and calcification. Under microscopy, a great number of growing cells could be seen and elastic and collagen fibers were abundant. Conclusion: Cultured self-derived endothelial and smooth muscle cells could be used as seeding cells and heterogeneous acellularized matrix could be used as scaffold in constructing tissue-engineered graft.
基金The authors are thankful to the Foundation for Research Support from the State of Sao Paulo(Fundacao de Amparoà Pesquisa do Estado de Sao Paulo-FAPESP)for the financial support needed for the execution of this research project(grant number:07/52027-9).
文摘Arterial embolization of myomas (AEM) is an established option for the conservative treatment of uterine leio-myomas;it treats all present uterine nodules at once, is less invasive than other procedures and effective in controlling symptoms, and does not require long term hospitalizations. Nevertheless, the potential impact on endometrial morphological and functional outcomes after the procedure is still controversial based on reports of endometritis or eventual transient ischemia. This study evaluated endometrial reorganization in uterine leiomyoma patients, before and after AEM, through gene expression analyses of extracellular matrix and cytokines genes in theendometrial tissue. Eight patients with leiomyomas were evaluated before AEM and 6 months after. The examinations included transvaginal pelvic ultrasonography, dosing of the follicle-stimulating hormone, and endometrial biopsy during the second phase of the menstrual cycle. RNA was extracted from endometrial samples, cDNA was synthesized, and applied on PCR arrayTM plates to evaluate the expression of extracellular matrix (ECM) genes and cytokines and their receptors’ genes (CYT). The ECM overexpressed genes were MMP (1, 3, 10, 11, and 14), CTGF1, ICAM1, TBHS1, ITGA2, ITGA3, ITGB3, COL7A1, COL12A, SPP1, and TNC;ADAMTS8 was underexpressed. The CYT overexpressed genes were SPP1, BCL6, CXCL12, IL-8, and CEBPB;CXCL13 and CCL21 were underexpressed. The ECM results showed overexpression of proteases that are responsible for dysfunctions in the ECM, and of genes responsible for adhesion and membrane components. The CYT results showed overexpression of chemokines responsible for endometrial repair, and underexpression of cytokines involved in inflammatory processes in the endometrial tissue. AEM treatment did not negatively affect the endometrial function at 6 months after embolization. This study broadens the knowledge about using a procedure that is relevant to the treatment of leiomyomas and contributes to the establishment of future guidelines for the decision making process for physicians and patients.
文摘Song et al have reported a 100% success rate of acellular extracellular matrix (AEM) anal fistula plug in low fistula-in-ano. The results with this product in high fistula-in-ano are keenly awaited.
文摘Objective To establish a method for quantitative detection of the sulfate glycosaminoglycans ( GAG) content in extracellular matrix of in vitro cultured chondrocytes so as to evaluate the biological characteristics of epiphyseal, articular and rib chondrocytes. Methods Sulfate GAG content in extracellular matrix of three chondrocytes was measured by the modified dimethylmethylene blue (DMB) method. The changes of the toluidine blue (TB) stain of chondrocytes were observed by light microscope. Results Primary chondrocytes had the highest content of sulfate GAG in the extracellular matrix, ie, epiphyseal chondrocytes reached ( 70. 12 ± 7. 72 )μg/cm2, articular chondrocytes (92.00 ± 10.15) μg/cm2 and rib chondrocytes (80.61 ± 11. 40) μg/cm2, respectively. On the third pasage chondrocytes, epiphyceal chondrocytes decreased to (53.27 ± 9. 50 ) μg/cm2, articular chondrocytes to (63.88 ± 11.92) μg/cm2 and rib chondrocytes to (58.94 ±8.21) μg/cm2, respectively. The change of TB in every passage
