Mesenchymal stem cells from osteoporotic patients produce a type I collagen-deficient extracellular matrix favoring adipogenic differentiation

Interactions between stem cells and extracellular matrix (ECM) are requisite for inducing lineage-specific differentiation and maintaining biological functions of mesenchymal stem cells by providing a composite set of chemical and structural signals. Here we investigated if cell-deposited ECM mimickedin vivoliver's stem cell microenvironment and facilitated hepatogenic maturation. Decellularization process preserved the fibrillar microstructure and a mix of matrix proteins in cell-deposited ECM, such as type I collagen, type III collagen, fibronectin, and laminin that were identical to those found in native liver. Compared with the cells on tissue culture polystyrene (TCPS), bone marrow mesenchymal stem cells (BM-MSCs) cultured on cell-deposited ECM showed a spindle-like shape, a robust proliferative capacity, and a suppressed level of intracellular reactive oxygen species, accompanied with upregulation of two superoxide dismutases. Hepatocyte-like cells differentiated from BM-MSCs on ECM were determined with a more intensive staining of glycogen storage, an elevated level of urea biosynthesis, and higher expressions of hepatocyte-specific genes in contrast to those on TCPS. These results demonstrate that cell-deposited ECM can be an effective method to facilitate hepatic maturation of BM-MSCs and promote stem-cell-based liver regenerative medicine.

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Basement membrane proteins modulate cell migration on bovine pericardium extracellular matrix scaffold

Scientific Reports ◽

10.1038/s41598-021-84161-5 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Qi Xing ◽

Mojtaba Parvizi ◽

Manuela Lopera Higuita ◽

Leigh G. Griffiths

Keyword(s):

Stem Cells ◽

Monoclonal Antibody ◽

Extracellular Matrix ◽

Mesenchymal Stem Cells ◽

Cell Migration ◽

Basement Membrane ◽

Type I Collagen ◽

Human Mesenchymal Stem Cells ◽

Bovine Pericardium ◽

Type I

AbstractNative bovine pericardium (BP) exhibits anisotropy of its surface ECM niches, with the serous surface (i.e., parietal pericardium) containing basement membrane components (e.g., Laminin, Col IV) and the fibrous surface (i.e., mediastinal side) being composed primarily of type I collagen (Col I). Native BP surface ECM niche anisotropy is preserved in antigen removed BP (AR-BP) extracellular matrix (ECM) scaffolds. By exploiting sideness (serous or fibrous surface) of AR-BP scaffolds, this study aims to determine the mechanism by which ECM niche influences human mesenchymal stem cells (hMSCs) migration. Human mesenchymal stem cells (hMSC) seeding on serous surface promoted more rapid cell migration than fibrous surface seeding. Gene analysis revealed that expression of integrin α3 and α11 were increased in cells cultured on serous surface compared to those on the fibrous side. Monoclonal antibody blockade of α3β1 (i.e., laminin binding) inhibited early (i.e. ≤ 6 h) hMSC migration following serous seeding, while having no effect on migration of cells on the fibrous side. Blockade of α3β1 resulted in decreased expression of integrin α3 by cells on serous surface. Monoclonal antibody blockade of α11β1 (i.e., Col IV binding) inhibited serous side migration at later time points (i.e., 6–24 h). These results confirmed the role of integrin α3β1 binding to laminin in mediating early rapid hMSCs migration and α11β1 binding to Col IV in mediating later hMSCs migration on the serous side of AR-BP, which has critical implications for rate of cellular monolayer formation and use of AR-BP as blood contacting material for clinical applications.

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APPLICATION OF DENDRIMER/PLASMID hBMP-2 COMPLEXES LOADED INTO β-TCP/COLLAGEN SCAFFOLD IN THE TREATMENT OF FEMORAL DEFECTS IN RATS

Biomedical Engineering Applications Basis and Communications ◽

10.4015/s1016237214500057 ◽

2014 ◽

Vol 26 (01) ◽

pp. 1450005 ◽

Cited By ~ 1

Author(s):

Tingwei Bao ◽

Huiming Wang ◽

Wentao Zhang ◽

Xuefeng Xia ◽

Jiabei Zhou ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Type I Collagen ◽

Cell Attachment ◽

Collagen Scaffold ◽

Bone Defects ◽

Bone Morphogenetic Protein 2 ◽

Porous Scaffolds ◽

Type I

Purpose: Plasmid loading into scaffolds to enhance sustained release of growth factors is an important focus of regenerative medicine. The aim of this study was to build gene-activated matrices (GAMs) and examine the bone augmentation properties. Methods: Generation 5 polyamidoamine dendrimers (G5 dPAMAM)/plasmid recombinant human bone morphogenetic protein-2 (rhBMP-2) complexes were immobilized into beta-tricalcium phosphate (β-TCP)/type I collagen porous scaffolds. After cultured with rat mesenchymal stem cells (rMSCs), transfection efficiencies were examined. The secretion of rhBMP-2 and alkaline phosphatase (ALP) were detected to evaluate the osteogenic properties. Scanning electron microscopy (SEM) was used to observe attachment and proliferation. Moreover, we applied these GAMs directly into freshly created segmental bone defects in rat femurs, and their osteogenic efficiencies were evaluated. Results: Released plasmid complexes were transfected into stem cells and were expressed, which caused osteogenic differentiations of rat mesenchymal stem cells (rMSCs). SEM analysis showed excellent cell attachment. Bioactivity of plasmid rhBMP-2 was maintained in vivo, and the X-ray observation, histological analysis and immunohistochemistry (IHC) of bone tissue demonstrated that the bone healing in segmental femoral defects was enhanced by implantation of GAMs. Conclusions: Such biomaterials offer therapeutic opportunities in critical-sized bone defects.

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Exogenous type I collagen facilitates osteogenic differentiation and acts as a substrate for mineralization of rat marrow mesenchymal stem cells in vitro

Biochemical and Biophysical Research Communications ◽

10.1016/j.bbrc.2006.01.059 ◽

2006 ◽

Vol 341 (4) ◽

pp. 1029-1035 ◽

Cited By ~ 47

Author(s):

Takanori Kihara ◽

Motohiro Hirose ◽

Akira Oshima ◽

Hajime Ohgushi

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Osteogenic Differentiation ◽

Type I Collagen ◽

Type I ◽

Marrow Mesenchymal Stem Cells

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Type I collagen promotes proliferation and osteogenesis of human mesenchymal stem cells via activation of ERK and Akt pathways

Journal of Biomedical Materials Research Part A ◽

10.1002/jbm.a.32693 ◽

2010 ◽

Vol 9999A ◽

pp. NA-NA ◽

Cited By ~ 11

Author(s):

Kuo-Shu Tsai ◽

Shou-Yen Kao ◽

Chien-Yuan Wang ◽

Yng-Jiin Wang ◽

Jung-Pan Wang ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Type I Collagen ◽

Human Mesenchymal Stem Cells ◽

Type I

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TGF-β Enhances the Integrin α2β1-Mediated Attachment of Mesenchymal Stem Cells to Type I Collagen

Stem Cells and Development ◽

10.1089/scd.2009.0208 ◽

2010 ◽

Vol 19 (5) ◽

pp. 645-656 ◽

Cited By ~ 26

Author(s):

Katrin Warstat ◽

Diana Meckbach ◽

Michaela Weis-Klemm ◽

Anita Hack ◽

Gerd Klein ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Type I Collagen ◽

Type I

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3D Biomimetic Scaffold for Growth Factor Controlled Delivery: An In-Vitro Study of Tenogenic Events on Wharton’s Jelly Mesenchymal Stem Cells

Pharmaceutics ◽

10.3390/pharmaceutics13091448 ◽

2021 ◽

Vol 13 (9) ◽

pp. 1448

Author(s):

Maria Camilla Ciardulli ◽

Joseph Lovecchio ◽

Pasqualina Scala ◽

Erwin Pavel Lamparelli ◽

Tina Patricia Dale ◽

...

Keyword(s):

Gene Expression ◽

Stem Cells ◽

Mesenchymal Stem Cells ◽

Type I Collagen ◽

In Vitro Study ◽

Wharton’S Jelly ◽

Controlled Delivery ◽

Type I ◽

Wharton's Jelly

The present work described a bio-functionalized 3D fibrous construct, as an interactive teno-inductive graft model to study tenogenic potential events of human mesenchymal stem cells collected from Wharton’s Jelly (hWJ-MSCs). The 3D-biomimetic and bioresorbable scaffold was functionalized with nanocarriers for the local controlled delivery of a teno-inductive factor, i.e., the human Growth Differentiation factor 5 (hGDF-5). Significant results in terms of gene expression were obtained. Namely, the up-regulation of Scleraxis (350-fold, p ≤ 0.05), type I Collagen (8-fold), Decorin (2.5-fold), and Tenascin-C (1.3-fold) was detected at day 14; on the other hand, when hGDF-5 was supplemented in the external medium only (in absence of nanocarriers), a limited effect on gene expression was evident. Teno-inductive environment also induced pro-inflammatory, (IL-6 (1.6-fold), TNF (45-fold, p ≤ 0.001), and IL-12A (1.4-fold)), and anti-inflammatory (IL-10 (120-fold) and TGF-β1 (1.8-fold)) cytokine expression upregulation at day 14. The presented 3D construct opens perspectives for the study of drug controlled delivery devices to promote teno-regenerative events.

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Presence of ROS in Inflammatory Environment of Peri-Implantitis Tissue: In Vitro and In Vivo Human Evidence

Journal of Clinical Medicine ◽

10.3390/jcm9010038 ◽

2019 ◽

Vol 9 (1) ◽

pp. 38

Author(s):

Eitan Mijiritsky ◽

Letizia Ferroni ◽

Chiara Gardin ◽

Oren Peleg ◽

Alper Gultekin ◽

...

Keyword(s):

Stem Cells ◽

Extracellular Matrix ◽

Mesenchymal Stem Cells ◽

Human Model ◽

Inflammatory Cell Infiltrate ◽

Mitochondrial Activity ◽

Type I ◽

Biological Organization ◽

Extracellular Matrix Components ◽

Matrix Components

Analyses of composition, distribution of cellular and extracellular matrix components, and molecular analysis of mitochondria related genes of bone loss in the presence of inflammatory environment in humans was the aim of the present project. As a human model we chose peri-implantitis. Morphological analyses were performed by means classical histological, immunohistochemical, and SEM (scanning electron miscroscopy) test. Gene expression analysis was performed to evaluate epithelium maturation, collagen fiber production, and genes related to mitochondrial activity. It was found that a well-defined keratinocyte epithelium was present on the top of all specimens; a distinct basal lamina was present, as well as desmosomes and autophagic processes related to the maturation of keratinocytes. Under this epithelium, a full inflammatory cell infiltrate was present for about 60% of the represented by plasma cells. Collagen type I fibers were present mainly in the form of fragmented cord tissue without cells. A different distribution of blood vessels was also present from the apical to the most coronal portion of the specimens. High levels of genes related to oxidative stress were present, as well as the activation of genes related to the loss of ability of osteogenic commitment of Mesenchymal stem cells into osteoblasts. Our study suggests that peri-implantitis lesions exhibit a well defined biological organization not only in terms of inflammatory cells but also on vessel and extracellular matrix components even if no difference in the epithelium is evident, and that the presence of reactive oxygen species (ROS) related to the inflammatory environment influences the correct commitment of Mesenchymal stem cells.

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Abstract 129: Human Mesenchymal Stem Cells Augment Contractile Function of Human Engineered Cardiac Tissues

Circulation Research ◽

10.1161/res.113.suppl_1.a129 ◽

2013 ◽

Vol 113 (suppl_1) ◽

Author(s):

Timothy Cashman ◽

Irene C Turnbull ◽

Ioannis Karakikes ◽

Jose Da Silva ◽

Joshua M Hare ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Type I Collagen ◽

High Efficiency ◽

Contractile Function ◽

Embryonic Stem ◽

Type I ◽

Cross Sectional ◽

Cardiac Tissues ◽

Twitch Force

Mesenchymal stem cells (MSC) have demonstrated efficacy for improving cardiomyocyte (CM) function in vitro, in vivo and in clinical trials, but the mechanism of this enhancement remains elusive. The objective of this study was to test the hypothesis that human engineered cardiac tissues (hECT) offer a viable model system to investigate the effects of human MSC on CM contractile function. Human CM (hCM) were produced from embryonic stem cells (hESC, H7 line) using a small-molecule based differentiation approach. Blebbistatin and BMP4 were added to hESC suspended in StemPro34 differentiation media for 24 h, followed by BMP4 and Activin A to day 4.5, followed by addition of IWR-1 Wnt inhibitor for at least 4 days. To create hECT, approximately 1 million hCM were mixed with 2.0 mg/ml bovine type I collagen and 0.9 mg/ml Matrigel, and pipetted into a mold fabricated from polydimethylsiloxane with integrated cantilever end-posts. To model hMSC cell therapy, two types of hECT were created: hCM-only control hECT, and hMSC-CM hybrid hECT containing hCM mixed with 5-10% of human bone marrow-derived MSC. Over several days in culture, the hECT self-assembled and started beating; end-post deflection was tracked in real time to compute twitch force using beam theory. Human CMs were produced with high efficiency (>70% cTnT+) with a predominantly ventricular phenotype (MLC2v+). Resulting hECTs exhibited spontaneous beating (1.3±0.4 Hz), cellular alignment, registered sarcomeres, and expression of cardiac specific genes cTnT, α-MHC, β-MHC and SERCA2a. After 11±2 days in culture, developed stress (force/area) was over 10-fold higher in hMSC-CM hybrid tissues (0.27±0.048 mN/mm 2 ) compared to hCM-only controls (0.02±0.006 mN/mm 2 ; p=0.04, n=5 per group). This reflected significantly greater twitch force (0.11±0.004 mN vs 0.033±0.016 mN, p=0.016) and smaller cross-sectional area (0.19±0.12 mm 2 vs 0.49±0.10 mm 2 ; p=0.003) in hMSC-CM hybrid vs hCM-only hECT. In conclusion, human ECT offer a novel system to study MSC-CM interactions. The findings suggest hMSC supplementation improves contractility compared to CM-only hECT. Investigating the mechanisms of hMSC-mediated enhancement of hECT function may yield insights into MSC-based therapies for cardiac regeneration.

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