Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 5 de 5
Filter
Add more filters










Database
Language
Publication year range
1.
Cells ; 10(11)2021 11 11.
Article in English | MEDLINE | ID: mdl-34831345

ABSTRACT

Adipose-derived stem cells (ASCs) are an abundant and easily accessible multipotent stem cell source with potential application in smooth muscle regeneration strategies. In 3D collagen hydrogels, we investigated whether sustained release of growth factors (GF) PDGF-AB and TGF-ß1 from GF-loaded microspheres could induce a smooth muscle cell (SMC) phenotype in ASCs, and if the addition of uniaxial cyclic stretch could enhance the differentiation level. This study demonstrated that the combination of cyclic stretch and GF release over time from loaded microspheres potentiated the differentiation of ASCs, as quantified by protein expression of early to late SMC differentiation markers (SMA, TGLN and smooth muscle MHC). The delivery of GFs via microspheres produced large ASCs with a spindle-shaped, elongated SMC-like morphology. Cyclic strain produced the largest, longest, and most spindle-shaped cells regardless of the presence or absence of growth factors or the growth factor delivery method. Protein expression and cell morphology data confirmed that the sustained release of GFs from GF-loaded microspheres can be used to promote the differentiation of ASCs into SMCs and that the addition of uniaxial cyclic stretch significantly enhances the differentiation level, as quantified by intermediate and late SMC markers and a SMC-like elongated cell morphology.


Subject(s)
Adipose Tissue/cytology , Intercellular Signaling Peptides and Proteins/pharmacology , Myocytes, Smooth Muscle/cytology , Stem Cells/cytology , Stress, Mechanical , Adult , Biomarkers/metabolism , Bioreactors , Cell Differentiation/drug effects , Cell Shape/drug effects , Cells, Cultured , Cross-Linking Reagents/chemistry , Gelatin/chemistry , Gels/chemistry , Humans , Iridoids/chemistry , Microspheres , Myocytes, Smooth Muscle/drug effects , Myocytes, Smooth Muscle/metabolism , Phenotype , Stem Cells/drug effects , Stem Cells/metabolism
2.
Sci Rep ; 7(1): 6640, 2017 07 26.
Article in English | MEDLINE | ID: mdl-28747783

ABSTRACT

Stem cells have been predicted to improve disease outcomes and patient lives. Steering stem cell fate - through controlling cell shape - may substantially accelerate progress towards this goal. As mesenchymal stromal cells (MSCs) are continuously exposed in vivo to a dynamically changing biomechanical environment, we hypothesized that exogenous forces can be applied for engineering a variety of significantly different MSC shapes. We applied specific cyclic stretch regimens to human MSCs and quantitatively measured the resulting cell shape, alignment, and expression of smooth muscle (SMC) differentiation markers, as those have been associated with elongated morphology. As proof of principle, a range of different shapes, alignments, and correlating SMC marker levels were generated by varying strain, length, and repetition of stretch. However, the major determinant of biomechanically engineering cellular shape was the repetition of a chosen stretch regimen, indicating that the engineered shape and associated differentiation were complex non-linear processes relying on sustained biomechanical stimulation. Thus, forces are key regulators of stem cell shape and the targeted engineering of specific MSC shapes through biomechanical forces represents a novel mechanobiology concept that could exploit naturally occurring in vivo forces for improving stem cell fate in clinical regenerative therapies.


Subject(s)
Cell Culture Techniques/methods , Cell Shape , Cytological Techniques/methods , Mesenchymal Stem Cells/cytology , Metabolic Engineering/methods , Aged , Aged, 80 and over , Cells, Cultured , Female , Humans , Male , Middle Aged , Stress, Mechanical
3.
J Tissue Eng Regen Med ; 11(12): 3508-3522, 2017 12.
Article in English | MEDLINE | ID: mdl-28371409

ABSTRACT

Controlling mesenchymal stromal cell (MSC) shape is a novel method for investigating and directing MSC behaviour in vitro. it was hypothesized that specifigc MSC shapes can be generated by using stiffness-defined biomaterial surfaces and by applying cyclic tensile forces. Biomaterials used were thin and thick silicone sheets, fibronectin coating, and compacted collagen type I sheets. The MSC morphology was quantified by shape descriptors describing dimensions and membrane protrusions. Nanoscale stiffness was measured by atomic force microscopy and the expression of smooth muscle cell (SMC) marker genes (ACTA2, TAGLN, CNN1) by quantitative reverse-transcription polymerase chain reaction. Cyclic stretch was applied with 2.5% or 5% amplitudes. Attachment to biomaterials with a higher stiffness yielded more elongated MSCs with fewer membrane protrusions compared with biomaterials with a lower stiffness. For cyclic stretch, compacted collagen sheets were selected, which were associated with the most elongated MSC shape across all investigated biomaterials. As expected, cyclic stretch elongated MSCs during stretch. One hour after cessation of stretch, however, MSC shape was rounder again, suggesting loss of stretch-induced shape. Different shape descriptor values obtained by different stretch regimes correlated significantly with the expression levels of SMC marker genes. Values of approximately 0.4 for roundness and 3.4 for aspect ratio were critical for the highest expression levels of ACTA2 and CNN1. Thus, specific shape descriptor values, which can be generated using biomaterial-associated stiffness and tensile forces, can serve as a template for the induction of specific gene expression levels in MSC. Copyright © 2017 John Wiley & Sons, Ltd.


Subject(s)
Biocompatible Materials/pharmacology , Cell Shape , Mesenchymal Stem Cells/cytology , Tensile Strength , Animals , Biomarkers/metabolism , Biomechanical Phenomena , Cell Adhesion/drug effects , Cell Shape/drug effects , Gene Expression Regulation/drug effects , Humans , Mesenchymal Stem Cells/drug effects , Myocytes, Smooth Muscle/cytology , Myocytes, Smooth Muscle/drug effects , Myocytes, Smooth Muscle/metabolism , Rats , Time Factors
4.
Sci Rep ; 6: 35840, 2016 10 24.
Article in English | MEDLINE | ID: mdl-27775041

ABSTRACT

Using matrix elasticity and cyclic stretch have been investigated for inducing mesenchymal stromal cell (MSC) differentiation towards the smooth muscle cell (SMC) lineage but not in combination. We hypothesized that combining lineage-specific stiffness with cyclic stretch would result in a significantly increased expression of SMC markers, compared to non-stretched controls. First, we generated dense collagen type I sheets by mechanically compressing collagen hydrogels. Atomic force microscopy revealed a nanoscale stiffness range known to support myogenic differentiation. Further characterization revealed viscoelasticity and stable biomechanical properties under cyclic stretch with >99% viable adherent human MSC. MSCs on collagen sheets demonstrated a significantly increased mRNA but not protein expression of SMC markers, compared to on culture flasks. However, cyclic stretch of MSCs on collagen sheets significantly increased both mRNA and protein expression of α-smooth muscle actin, transgelin, and calponin versus plastic and non-stretched sheets. Thus, lineage-specific stiffness and cyclic stretch can be applied together for inducing MSC differentiation towards SMCs without the addition of recombinant growth factors or other soluble factors. This represents a novel stimulation method for modulating the phenotype of MSCs towards SMCs that could easily be incorporated into currently available methodologies to obtain a more targeted control of MSC phenotype.


Subject(s)
Cell Culture Techniques/methods , Collagen Type I/chemistry , Mesenchymal Stem Cells/cytology , Muscle, Smooth/cytology , Actins/genetics , Biomarkers/metabolism , Bone Marrow Cells/cytology , Calcium-Binding Proteins/genetics , Cell Differentiation/physiology , Cells, Cultured , Gene Expression Regulation , Humans , Mesenchymal Stem Cells/physiology , Microfilament Proteins/genetics , Microscopy, Atomic Force , Muscle Proteins/genetics , Phenotype , Tubulin/metabolism , Calponins
5.
Food Chem Toxicol ; 71: 68-73, 2014 Sep.
Article in English | MEDLINE | ID: mdl-24928766

ABSTRACT

The toxic effects of acrylamide on cytoskeletal integrity and ion channel balance is well-established in many cell types, but there has been little examination regarding the effects of acrylamide on primary cardiomyocytes, despite the importance of such components in their function. Furthermore, acrylamide toxicity is generally examined using concentrations higher than those found in vivo under starch-rich diets. Accordingly, we sought to characterize the dose-dependent effects of acrylamide on various properties, including cell morphology, contraction patterns, and junctional connexin 43 staining, in primary cardiomyocytes. We show that several days exposure to 1-100 µM acrylamide resulted in altered morphology, irregular contraction patterns, and an increase in the amount of immunoreactive signal for connexin 43 at cell junctions. We conclude that dietary levels of acrylamide may alter cellular function with prolonged exposure, in primary cardiomyocytes.


Subject(s)
Acrylamide/toxicity , Diet , Myocytes, Cardiac/drug effects , Animals , Connexin 43/metabolism , Dose-Response Relationship, Drug , Rats , Rats, Wistar
SELECTION OF CITATIONS
SEARCH DETAIL
...