scholarly journals Human Amniotic Membrane Enriched with Urinary Bladder Fibroblasts Promote the Re-Epithelization of Urothelial Injury

2020 ◽  
Vol 29 ◽  
pp. 096368972094666
Author(s):  
Urška Dragin Jerman ◽  
Peter Veranič ◽  
Tina Cirman ◽  
Mateja Erdani Kreft
Keyword(s):  
Extracellular Matrix ◽  
Amniotic Membrane ◽  
Wound Dressing ◽  
Ex Vivo ◽  
Cell Junctions ◽  
Human Amniotic Membrane ◽  
Urothelial Cells ◽  
Ex Vivo Model ◽  
Bladder Tissue ◽  
Extracellular Matrix Components

Culturing cells in three-dimensional systems that include extracellular matrix components and different cell types mimic the native tissue and as such provide much more representative results than conventional two-dimensional cell cultures. In order to develop biomimetic bladder tissue in vitro, we used human amniotic membrane (AM) extracellular matrix as a scaffold for bladder fibroblasts (BFs) and urothelial cells. Our aims were to evaluate the integration of BFs into the AM stroma, to assess the differentiation of the urothelium on BFs-enriched AM scaffolds, and to evaluate the AM as a urothelial wound dressing. First, to achieve the optimal integration of BFs into AM stroma, different intact and de- epithelialized AM (dAM) scaffolds were tested. BFs secreted matrix metalloproteinase (MMP)-1 and MMP-2 and integrated into the stroma of all types of AM scaffolds. Second, to establish urothelial tissue equivalent, urothelial cells were seeded on dAM scaffolds enriched with BFs. The BFs in the stroma of the AM scaffolds promoted (1) the proliferation of urothelial cells, (2) the attachment of urothelial cells on AM basal lamina with hemidesmosomes, and (3) development of multilayered urothelium with expressed uroplakins and well-developed cell junctions. Third, we established an ex vivo model of the injured bladder to evaluate the dAM as a wound dressing for urothelial full-thickness injury. dAM acted as a promising wound dressing since it enabled rapid re-epithelization of urothelial injury and integrated into the bladder tissue. Herein, the developed urothelial tissue equivalents enable further mechanistic studies of bladder epithelial–mesenchymal interactions, and they could be applied as biomimetic models for preclinical testing of newly developed drugs. Moreover, we could hypothesize that AM may be suitable as a dressing of the wound that occurs during transurethral resection of bladder tumor, since it could diminish the possibility of tumor recurrence, by promoting the rapid re-epithelization of the urothelium.

scholarly journals Precision-Cut Kidney Slices as a Tool to Understand the Dynamics of Extracellular Matrix Remodeling in Renal Fibrosis

2016 ◽  
Vol 11 ◽  
pp. BMI.S38439 ◽  
Author(s):  
Federica Genovese ◽  
Zsolt S. Kàrpàti ◽  
Signe H. Nielsen ◽  
Morten A. Karsdal
Keyword(s):  
Extracellular Matrix ◽  
Collagen Type ◽  
Interstitial Fibrosis ◽  
Ex Vivo ◽  
Collagen Type I ◽  
Extracellular Matrix Remodeling ◽  
Type I ◽  
Matrix Remodeling ◽  
Renal Interstitial Fibrosis ◽  
Ex Vivo Model

The aim of this study was to set up an ex vivo model for renal interstitial fibrosis in order to investigate the extracellular matrix (ECM) turnover profile in the fibrotic kidney. We induced kidney fibrosis in fourteen 12-week-old male Sprague Dawley rats by unilateral ureteral obstruction (UUO) surgery of the right ureter. The left kidney (contralateral) was used as internal control. Six rats were sham operated and used as the control group. Rats were terminated two weeks after the surgery; the kidneys were excised and precision-cut kidney slices (PCKSs) were cultured for five days in serum-free medium. Markers of collagen type I formation (P1NP), collagen type I and III degradation (C1M and C3M), and α-smooth muscle actin (αSMA) were measured in the PCKS supernatants by enzyme-linked immunosorbent assay. P1NP, C1M, C3M, and α-SMA were increased up to 2- to 13-fold in supernatants of tissue slices from the UUO-ligated kidneys compared with the contralateral kidneys ( P < 0.001) and with the kidneys of sham-operated animals ( P < 0.0001). The markers could also reflect the level of fibrosis in different animals. The UUO PCKS ex vivo model provides a valuable translational tool for investigating the extracellular matrix remodeling associated with renal interstitial fibrosis.

scholarly journals MT1-MMP–dependent neovessel formation within the confines of the three-dimensional extracellular matrix

2004 ◽  
Vol 167 (4) ◽  
pp. 757-767 ◽  
Author(s):  
Tae-Hwa Chun ◽  
Farideh Sabeh ◽  
Ichiro Ota ◽  
Hedwig Murphy ◽  
Kevin T. McDonagh ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Endothelial Cells ◽  
Cell Surface ◽  
Type I Collagen ◽  
Ex Vivo ◽  
Three Dimensional ◽  
Collagenolytic Activity ◽  
Type I ◽  
Ex Vivo Model

During angiogenesis, endothelial cells initiate a tissue-invasive program within an interstitial matrix comprised largely of type I collagen. Extracellular matrix–degradative enzymes, including the matrix metalloproteinases (MMPs) MMP-2 and MMP-9, are thought to play key roles in angiogenesis by binding to docking sites on the cell surface after activation by plasmin- and/or membrane-type (MT) 1-MMP–dependent processes. To identify proteinases critical to neovessel formation, an ex vivo model of angiogenesis has been established wherein tissue explants from gene-targeted mice are embedded within a three-dimensional, type I collagen matrix. Unexpectedly, neither MMP-2, MMP-9, their cognate cell-surface receptors (i.e., β3 integrin and CD44), nor plasminogen are essential for collagenolytic activity, endothelial cell invasion, or neovessel formation. Instead, the membrane-anchored MMP, MT1-MMP, confers endothelial cells with the ability to express invasive and tubulogenic activity in a collagen-rich milieu, in vitro or in vivo, where it plays an indispensable role in driving neovessel formation.

scholarly journals Adipose-Derived Stromal Cells and Mineralized Extracellular Matrix Delivery by a Human Decellularized Amniotic Membrane in Periodontal Tissue Engineering

Membranes ◽  
2021 ◽  
Vol 11 (8) ◽  
pp. 606
Author(s):  
Dilcele Silva Moreira Dziedzic ◽  
Bassam Felipe Mogharbel ◽  
Ana Carolina Irioda ◽  
Priscila Elias Ferreira Stricker ◽  
Maiara Carolina Perussolo ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Extracellular Matrix ◽  
Bone Tissue ◽  
Amniotic Membrane ◽  
Stromal Cells ◽  
Cell Attachment ◽  
Periodontal Regeneration ◽  
Human Amniotic Membrane ◽  
Micro Computed Tomography ◽  
Adipose Derived Stromal Cells

Periodontitis is a prevalent disease characterized by the loss of periodontal supporting tissues, bone, periodontal ligament, and cementum. The application of a bone tissue engineering strategy with Decellularized Human Amniotic Membrane (DAM) with adipose-derived stromal cells (ASCs) has shown to be convenient and valuable. This study aims to investigate the treatments of a rat periodontal furcation defect model with DAM, ASCs, and a mineralized extracellular matrix (ECM). Rat ASCs were expanded, cultivated on DAM, and with a bone differentiation medium for four weeks, deposited ECM on DAM. Periodontal healing for four weeks was evaluated by micro-computed tomography and histological analysis after treatments with DAM, ASCs, and ECM and compared to untreated defects on five consecutive horizontal levels, from gingival to apical. The results demonstrate that DAM preserves its structure during cultivation and healing periods, supporting cell attachment, permeation, bone deposition on DAM, and periodontal regeneration. DAM and DAM+ASCs enhance bone healing compared to the control on the gingival level. In conclusion, DAM with ASC or without cells and the ECM ensures bone tissue healing. The membrane supported neovascularization and promoted osteoconduction.

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