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https://www.readbyqxmd.com/read/28343003/essential-design-considerations-for-the-resazurin-reduction-assay-to-noninvasively-quantify-cell-expansion-within-perfused-extracellular-matrix-scaffolds
#1
Joseph S Uzarski, Michael D DiVito, Jason A Wertheim, William M Miller
Precise measurement of cellularity within bioartificial tissues and extracellular matrix (ECM) scaffolds is necessary to augment rigorous characterization of cellular behavior, as accurate benchmarking of tissue function to cell number allows for comparison of data across experiments and between laboratories. Resazurin, a soluble dye that is reduced to highly fluorescent resorufin in proportion to the metabolic activity of a cell population, is a valuable, noninvasive tool to measure cell number. We investigated experimental conditions in which resazurin reduction is a reliable indicator of cellularity within three-dimensional (3D) ECM scaffolds...
February 16, 2017: Biomaterials
https://www.readbyqxmd.com/read/28342324/short-bursts-of-cyclic-mechanical-compression-modulate-tissue-formation-in-a-3d-hybrid-scaffold
#2
M Brunelli, C M Perrault, D Lacroix
Among the cues affecting cells behaviour, mechanical stimuli are known to have a key role in tissue formation and mineralization of bone cells. While soft scaffolds are better at mimicking the extracellular environment, they cannot withstand the high loads required to be efficient substitutes for bone in vivo. We propose a 3D hybrid scaffold combining the load-bearing capabilities of polycaprolactone (PCL) and the ECM-like chemistry of collagen gel to support the dynamic mechanical differentiation of human embryonic mesodermal progenitor cells (hES-MPs)...
March 18, 2017: Journal of the Mechanical Behavior of Biomedical Materials
https://www.readbyqxmd.com/read/28342321/improvement-of-paracellular-transport-in-the-caco-2-drug-screening-model-using-protein-engineered-substrates
#3
Rebecca L DiMarco, Daniel R Hunt, Ruby E Dewi, Sarah C Heilshorn
The Caco-2 assay has achieved wide popularity among pharmaceutical companies in the past two decades as an in vitro method for estimation of in vivo oral bioavailability of pharmaceutical compounds during preclinical characterization. Despite its popularity, this assay suffers from a severe underprediction of the transport of drugs which are absorbed paracellularly, that is, which pass through the cell-cell tight junctions of the absorptive cells of the small intestine. Here, we propose that simply replacing the collagen I matrix employed in the standard Caco-2 assay with an engineered matrix, we can control cell morphology and hence regulate the cell-cell junctions that dictate paracellular transport...
March 18, 2017: Biomaterials
https://www.readbyqxmd.com/read/28339802/alterations-in-urinary-collagen-peptides-in-lupus-nephritis-subjects-correlate-with-renal-dysfunction-and-renal-histopathology
#4
Ru Wei, Benbo Gao, Fei Shih, Ann Ranger, Andrea Dearth, Harald Mischak, Justyna Siwy, Nicolas Wisniacki, Michelle Petri, Linda C Burkly
Background.: The excessive accumulation of extracellular matrix (ECM) in the renal tubulointerstitium is a key component of chronic renal damage in lupus nephritis (LN) and a critical determinant of the disease progression to renal failure. Detection of fibrosis requires renal biopsy and is therefore limited by high risks associated with an invasive procedure. This study explores whether a unique LN urinary peptidome can be identified and whether LN-specific alteration reflects the underlying fibrogenic process of altered ECM turnover...
February 23, 2017: Nephrology, Dialysis, Transplantation
https://www.readbyqxmd.com/read/28339082/effects-of-autophagy-and-endocytosis-on-the-activity-of-matrix-metalloproteinase%C3%A2-2-in-human-renal-proximal-tubular-cells-under-hypoxia
#5
Wenmin Yu, Zhi Wang, Yiping Li, Lei Liu, Jing Liu, Fenggan Ding, Xiaoyi Zhang, Zhengyuan Cheng, Pingsheng Chen
Tubulointerstitial fibrosis is characterized by tubular atrophy with basement membrane thickening and accumulation of interstitial extracellular matrix (ECM). A decrease in the activity of matrix metalloproteinase‑2 (MMP‑2) may promote this process. Although proximal tubular cells are sensitive to oxygen deprivation, whether cellular autophagy or endocytosis induced by hypoxia can alter the activity of MMP‑2 remains to be elucidated. The aim of the present study was to investigate whether autophagy and endocytosis induced by hypoxia can have an effect on the activity of MMP‑2 in HK‑2 cells...
March 22, 2017: Molecular Medicine Reports
https://www.readbyqxmd.com/read/28339064/extracellular-matrix-metabolism-disorder-induced-by-mechanical-strain-on-human-parametrial-ligament-fibroblasts
#6
Jie Min, Bingshu Li, Cheng Liu, Wenjun Guo, Shasha Hong, Jianming Tang, Li Hong
Pelvic organ prolapse (POP) is a global health problem that may seriously impact the quality of life of the sufferer. The present study aimed to investigate the potential mechanisms underlying alterations in extracellular matrix (ECM) metabolism in the pathogenesis of POP, by investigating the expression of ECM components in human parametrial ligament fibroblasts (hPLFs) subject to various mechanical strain loads. Fibroblasts derived from parametrial ligaments were cultured from patients with POP and without malignant tumors, who underwent vaginal hysterectomy surgery...
March 24, 2017: Molecular Medicine Reports
https://www.readbyqxmd.com/read/28338443/fluorescent-labeling-of-collagen-production-by-cells-for-noninvasive-imaging-of-extracellular-matrix-deposition
#7
Katie Bardsley, Ying Yang, Alicia J El Haj
Extracellular matrix (ECM) is an essential component of tissues and provides both integrity and biological cues for cells. Collagen is one of the major proteins found within the ECM and therefore is an essential component of all engineered tissues. Therefore, in this article, we present a method for the online real-time monitoring of collagen deposition in three-dimensional engineered constructs. This method revolves around modification of collagen through the addition of azide-L-proline to cell culture media...
March 24, 2017: Tissue Engineering. Part C, Methods
https://www.readbyqxmd.com/read/28338421/harnessing-the-osteogenicity-of-in-vitro-stem-cell-derived-mineralized-extracellular-matrix-as-3d-biotemplate-to-guide-bone-regeneration
#8
Yoke Chin Chai, Johanna Bolander, Ioannis Papantoniou, Jennifer Patterson, Jef Vleugels, Jan Schrooten, Frank P Luyten
Advanced biomaterials that are capable of guiding robust bone regeneration are highly demanded for translational therapy of bone defects or bone augmentation in clinics. One of the strategic approaches is to produce tissue engineering (TE) constructs that mediate bone regeneration by recapitulating the natural bone formation or healing process. In this study, we aimed at producing devitalized mineralized carriers with augmented bone forming capacity via a modified culture protocol (i.e., culture conditions with high calcium and/or phosphate concentrations) that first promotes cell growth and, subsequently, mineralized extracellular matrix (ECM) deposition by human periosteum-derived osteoprogenitor cells (hPDCs) on additive manufactured three-dimensional (3D) porous titanium (Ti)-based scaffolds...
March 24, 2017: Tissue Engineering. Part A
https://www.readbyqxmd.com/read/28338091/intermediate-filament-reorganization-dynamically-influences-cancer-cell-alignment-and-migration
#9
Andrew W Holle, Melih Kalafat, Adria Sales Ramos, Thomas Seufferlein, Ralf Kemkemer, Joachim P Spatz
The interactions between a cancer cell and its extracellular matrix (ECM) have been the focus of an increasing amount of investigation. The role of the intermediate filament keratin in cancer has also been coming into focus of late, but more research is needed to understand how this piece fits in the puzzle of cytoskeleton-mediated invasion and metastasis. In Panc-1 invasive pancreatic cancer cells, keratin phosphorylation in conjunction with actin inhibition was found to be sufficient to reduce cell area below either treatment alone...
March 24, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28337850/high-throughput-screening-of-vascular-endothelium-destructive-or-protective-microenvironments-cooperative-actions-of-extracellular-matrix-composition-stiffness-and-structure
#10
Yonghui Ding, Michael Floren, Wei Tan
Pathological modification of the subendothelial extracellular matrix (ECM) has closely been associated with endothelial activation and subsequent cardiovascular disease progression. To understand regulatory mechanisms of these matrix modifications, the majority of previous efforts have focused on the modulation of either chemical composition or matrix stiffness on 2D smooth surfaces without simultaneously probing their cooperative effects on endothelium function on in vivo like 3D fibrous matrices. To this end, a high-throughput, combinatorial microarray platform on 2D and 3D hydrogel settings to resemble the compositions, stiffness, and structure of healthy and diseased subendothelial ECM has been established, and further their respective and combined effects on endothelial attachment, proliferation, inflammation, and junctional integrity have been investigated...
March 24, 2017: Advanced Healthcare Materials
https://www.readbyqxmd.com/read/28337222/a-conditioned-medium-of-umbilical-cord-mesenchymal-stem-cells-overexpressing-wnt7a-promotes-wound-repair-and-regeneration-of-hair-follicles-in-mice
#11
Liang Dong, Haojie Hao, Jiejie Liu, Dongdong Ti, Chuan Tong, Qian Hou, Meirong Li, Jingxi Zheng, Gang Liu, Xiaobing Fu, Weidong Han
Mesenchymal stem cells (MSCs) can affect the microenvironment of a wound and thereby accelerate wound healing. Wnt proteins act as key mediators of skin development and participate in the formation of skin appendages such as hair. The mechanisms of action of MSCs and Wnt proteins on skin wounds are largely unknown. Here, we prepared a Wnt7a-containing conditioned medium (Wnt-CM) from the supernatant of cultured human umbilical cord-MSCs (UC-MSCs) overexpressing Wnt7a in order to examine the effects of this CM on cutaneous healing...
2017: Stem Cells International
https://www.readbyqxmd.com/read/28336462/the-molecular-mechanisms-of-androgen-receptor-in-nephrolithiasis
#12
Ziyu Fang, Yonghan Peng, Ling Li, Min Liu, Zeyu Wang, Shaoxiong Ming, Wenhui Zhang, Xiaofeng Gao
OBJECTIVES: This study aimed to investigate the molecular mechanisms of androgen receptor (AR) in nephrolithiasis. METHODS: Human kidney 2(HK-2) cells were transfected with Lentiviruses expressing AR (DEC-AR), shRNA targeting AR (sh-AR) or the empty vector control using the pLEX lentiviral vector system. The expression levels of AR were measured by qRT-PCR at 72h postinfection, and cells under different treatments were collected for microarray analysis. Differentially expressed genes (DEGs) were identified using Student's t-test...
March 21, 2017: Gene
https://www.readbyqxmd.com/read/28336453/a-bioinformatic-analysis-of-micrornas-role-in-osteoarthritis
#13
Lin Cong, Yue Zhu, Guanjun Tu
OBJECTIVE: To evaluate the underlying function of miRNAs in osteoarthritis (OA). DESIGN: A bioinformatic analysis of miRNAs-OA studies was completed in multiple databases. All identified articles were assessed using specific inclusion and exclusion criteria (Eligible case-control studies for the present study included those which investigated miRNAs differential expression in cartilage tissues and cells of OA and controls. Abstracts, case reports, conference presentations, editorials, and expert opinions were excluded...
March 21, 2017: Osteoarthritis and Cartilage
https://www.readbyqxmd.com/read/28335694/construction-of-three-dimensional-dermo-epidermal-skin-equivalents-using-cell-coating-technology-and-their-utilization-as-alternative-skin-for-permeation-studies-and-skin-irritation-tests
#14
Takami Akagi, Mayuka Nagura, Ayami Hiura, Hajime Kojima, Mitsuru Akashi
In vitro generated human skin equivalents are generating interest as promising tools in basic research, as alternatives to animal testing and for clinical applications in regenerative medicine. For prediction of skin irritation and corrosion, three-dimensional (3D) human skin equivalents consisting of differentiated human keratinocytes have been developed and some models have been internationally accepted. However, more delicate assessments using full-thickness skin models, such as skin sensitization tests cannot be performed because of the lack of a dermis containing fibroblasts or appendages...
March 23, 2017: Tissue Engineering. Part A
https://www.readbyqxmd.com/read/28335679/puerarin-attenuates-renal-fibrosis-by-reducing-oxidative-stress-induced-epithelial-cell-apoptosis-via-mapk-signal-pathways-in-vivo-and-in-vitro
#15
Xiangjun Zhou, Chen Bai, Xinbo Sun, Xiaoxin Gong, Yong Yang, Congbo Chen, Guang Shan, Qisheng Yao
Puerarin (PR) is an isoflavonoid isolated from the root of the plant Pueraria lobata and has been widely used in traditional Chinese herbal medicine for the treatment of various diseases. Oxidative stress and epithelial cell apoptosis play important roles in the renal fibrotic process. The present study aimed to determine whether or not PR inhibits renal fibrosis by reducing oxidative stress induced-epithelial cell apoptosis. In vivo, unilateral ureteral obstruction (UUO) induced renal fibrosis, and epithelial cell apoptosis...
November 2017: Renal Failure
https://www.readbyqxmd.com/read/28334523/transformable-nanomaterials-as-artificial-extracellular-matrix-for-inhibiting-tumor-invasion-and-metastasis
#16
Xiao-Xue Hu, Ping-Ping He, Guo-Bin Qi, Yu-Juan Gao, Yao-Xin Lin, Chao Yang, Pei-Pei Yang, Hongxun Hao, Lei Wang, Hao Wang
Tumor metastasis is one of the big challenges in cancer treatment and is often associated with high patient mortality. Until now, there is an agreement that tumor invasion and metastasis are related to degradation of extracellular matrix (ECM) by enzymes. Inspired by the formation of natural ECM and the in situ self-assembly strategy developed in our group, herein, we in situ constructed an artificial extracellular matrix (AECM) based on transformable Laminin (LN)-mimic peptide 1 (BP-KLVFFK-GGDGR-YIGSR) for inhibition of tumor invasion and metastasis...
March 23, 2017: ACS Nano
https://www.readbyqxmd.com/read/28333953/involvement-of-%C3%AE-and-%C3%AE-actin-isoforms-in-actin-cytoskeleton-organization-and-migration-abilities-of-bleb-forming-human-colon-cancer-cells
#17
Aleksandra Simiczyjew, Antonina Joanna Mazur, Ewelina Dratkiewicz, Dorota Nowak
Amoeboid movement is characteristic for rounded cells, which do not form strong adhesion contacts with the ECM and use blebs as migratory protrusions. It is well known that actin is the main component of mature forms of these structures, but the exact role fulfilled by non-muscle actin isoforms β- and γ- in bleb formation and migration of these cells is still not fully understood. The aim of this study was to establish the role of β- and γ-actin in migration of bleb-forming cancer cells using isoform-specific antibodies and expression of fluorescently tagged actin isoforms...
2017: PloS One
https://www.readbyqxmd.com/read/28333845/changes-in-the-extracellular-matrix-are-associated-with-the-development-of-serous-tubal-intraepithelial-carcinoma-into-high-grade-serous-carcinoma
#18
Sophieke C H A van der Steen, Johan Bulten, Koen K Van de Vijver, Toin H van Kuppevelt, Leon F A G Massuger
OBJECTIVE: The identification of a marker for early progression of preinvasive lesions into invasive pelvic high-grade serous carcinoma (HGSC) may provide novel handles for innovative screening and prevention strategies. The interplay between cancer cells and the extracellular matrix (ECM) is one of the main principles in cancer development and growth, but has been largely neglected in preinvasive lesions. This is the first study addressing the involvement of the ECM in the "step-by-step" transition of normal fallopian tube epithelium into preinvasive lesions, and eventually the progression of preinvasive lesions into invasive HGSC...
March 23, 2017: International Journal of Gynecological Cancer
https://www.readbyqxmd.com/read/28333195/genome-wide-genetic-analyses-highlight-mitogen-activated-protein-kinase-mapk-signaling-in-the-pathogenesis-of-endometriosis
#19
Outi Uimari, Nilufer Rahmioglu, Dale R Nyholt, Katy Vincent, Stacey A Missmer, Christian Becker, Andrew P Morris, Grant W Montgomery, Krina T Zondervan
STUDY QUESTION: Do genome-wide association study (GWAS) data for endometriosis provide insight into novel biological pathways associated with its pathogenesis? SUMMARY ANSWER: GWAS analysis uncovered multiple pathways that are statistically enriched for genetic association signals, analysis of Stage A disease highlighted a novel variant in MAP3K4, while top pathways significantly associated with all endometriosis and Stage A disease included several mitogen-activated protein kinase (MAPK)-related pathways...
February 9, 2017: Human Reproduction
https://www.readbyqxmd.com/read/28332281/collagen-type-xv-and-the-osteogenic-status
#20
Gina Lisignoli, Elisabetta Lambertini, Cristina Manferdini, Elena Gabusi, Letizia Penolazzi, Francesca Paolella, Marco Angelozzi, Veronica Casagranda, Roberta Piva
We have previously demonstrated that collagen type XV (ColXV) is a novel bone extracellular matrix (ECM) protein. It is well known that the complex mixture of multiple components present in ECM can help both to maintain stemness or to promote differentiation of stromal cells following change in qualitative characteristics or concentrations. We investigated the possible correlation between ColXV expression and mineral matrix deposition by human mesenchymal stromal cells (hMSCs) with different osteogenic potential and by osteoblasts (hOBs) that are able to grow in culture medium with or without calcium...
March 22, 2017: Journal of Cellular and Molecular Medicine
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