scholarly journals Pathological Process of Prompt Connection between Host and Donor Tissue Vasculature Causing Rapid Perfusion of the Engineered Donor Tissue after Transplantation

2018 ◽  
Vol 19 (12) ◽  
pp. 4102
Author(s):  
Sachiko Sekiya ◽  
Shunichi Morikawa ◽  
Taichi Ezaki ◽  
Tatsuya Shimizu
Keyword(s):  
Endothelial Cell ◽  
Cell Sheet ◽  
Three Dimensional ◽  
Blood Perfusion ◽  
Myocardial Cell ◽  
Donor Tissue ◽  
Cell Network ◽  
Engineered Tissue

The shortage of donors for transplantation therapy is a serious issue worldwide. Tissue engineering is considered a potential solution to this problem. Connection and perfusion in engineered tissues after transplantation is vital for the survival of the transplanted tissue, especially for tissues requiring blood perfusion to receive nutrients, such as the heart. A myocardial cell sheet containing an endothelial cell network structure was fabricated in vitro using cell sheet technology. Transplantation of the three-dimensional (3D) tissue by layering myocardial sheets could ameliorate ischemic heart disease in a rat model. The endothelial cell network in the 3D tissue was able to rapidly connect to host vasculature and begin perfusion within 24 h after transplantation. In this review, we compare and discuss the engineered tissue–host vasculature connection process between tissue engineered constructs with hydrogels and cell sheets by histological analysis. This review provides information that may be useful for further improvements of in vivo engineered tissue vascularization techniques.

Abstract 186: Hdac7-derived 7aa Peptide May Function as a Phosphorylation Carrier

2016 ◽  
Vol 36 (suppl_1) ◽  
Author(s):  
Junyao Yang ◽  
Wen Wang ◽  
Qian Wang ◽  
Lingfang Zeng
Keyword(s):  
Endothelial Cell ◽  
Vascular Endothelial Cell ◽  
Blood Perfusion ◽  
Open Reading Frames ◽  
In Vivo Studies ◽  
Buffer System ◽  
Vascular Progenitor Cells ◽  
Upstream Kinase

Background: Histone deacetylase 7 (HDAC7) belongs to class II HDAC family, playing a pivotal role in the maintenance of endothelium integrity. There are 8 splicing variants in mouse HDAC7 mRNAs. Within the 5’ terminal non-coding area of some variants, there exist some short open reading frames (sORFs). Whether these sORFs can be translated and their potential roles in cellular physiology remain unclear. Method and results: Our previous studies suggested that one mouse HDAC7 produced a 7aa peptide from the non-coding area. In this study, we demonstrated that one sORF encoding a 7 amino acids (aa)-peptide could be translated in response to vascular endothelial cell growth factor (VEGF) in vascular progenitor cells (VPCs). The 7aa-peptide (7A) could be phosphorylated at serine residue via MEKK1. Importantly, the phosphorylated 7aa-peptide (7Ap) could transfer the phosphorylation group to the Thr residue of the 14-3-3γ protein in a cell free in-gel buffer system. The in vitro functional analyses revealed that 7A enhanced VEGF-induced VPC migration and differentiation toward endothelial cell (EC) lineage, in which MEKK1 and 14-3-3γ served as upstream kinase and downstream effector respectively. Knockdown of either MEKK1 or 14-3-3γ attenuated VEGF-induced VPC migration and differentiation. Exogenous 7Ap could rescue VEGF effect in MEKK1 but not in 14-3-3γ knockdown cells. The in vivo studies showed that 7A especially 7Ap induced capillary vessel formation within matrigel plug assays, increased re-endothelialization and suppressed neointima formation in the femoral artery injury model, and promoted the foot blood perfusion recovery in the hindlimb ischemia model. Conclusion: These results indicate that the sORFs within the non-coding area can be translated under some circumstances and that the 7aa-peptide may play an important role in cellular processes like migration and differentiation via acting as a phosphorylation carrier. Significance: As a phosphorylation carrier, 7aa possesses therapeutic potentials in tackling angiogenesis related diseases.

Three-dimensional cell-dense constructs containing endothelial cell-networks are an effective tool for in vivo and in vitro vascular biology research

2010 ◽  
Vol 80 (3) ◽  
pp. 549-551 ◽  
Author(s):  
Sachiko Sekiya ◽  
Megumi Muraoka ◽  
Tadashi Sasagawa ◽  
Tatsuya Shimizu ◽  
Masayuki Yamato ◽  
...  
Keyword(s):  
Endothelial Cell ◽  
Three Dimensional ◽  
Vascular Biology ◽  
Cell Networks ◽  
Biology Research ◽  
Dimensional Cell

scholarly journals A Light Wand to Untangle the Myocardial Cell Network

2019 ◽  
Vol 2 (2) ◽  
pp. 34 ◽  
Author(s):  
Tania Zaglia ◽  
Anna Di Bona ◽  
Marco Mongillo
Keyword(s):  
Cardiac Arrhythmias ◽  
Myocardial Cell ◽  
Model Organisms ◽  
Cell Type ◽  
Cell Network ◽  
Selective Modulation ◽  
Targeted Expression ◽  
Myocardial Biology

The discovery of optogenetics has revolutionized research in neuroscience by providing the tools for noninvasive, cell-type selective modulation of membrane potential and cellular function in vitro and in vivo. Rhodopsin-based optogenetics has later been introduced in experimental cardiology studies and used as a tool to photoactivate cardiac contractions or to identify the sites, timing, and location most effective for defibrillating impulses to interrupt cardiac arrhythmias. The exploitation of cell-selectivity of optogenetics, and the generation of model organisms with myocardial cell type targeted expression of opsins has started to yield novel and sometimes unexpected notions on myocardial biology. This review summarizes the main results, the different uses, and the prospective developments of cardiac optogenetics.

Design and Fabrication of a Constant Shear Microfluidic Network for Tissue Engineering

2004 ◽  
Vol 820 ◽  
Author(s):  
E.J. Weinberg ◽  
J.T. Borenstein ◽  
M.R. Kaazempur-Mofrad ◽  
B. Orrick ◽  
J.P. Vacanti
Keyword(s):  
Tissue Engineering ◽  
Blood Flow ◽  
Shear Stress ◽  
Endothelial Cell ◽  
Dynamic Properties ◽  
Fluid Dynamic ◽  
Three Dimensional ◽  
Cell Behavior

AbstractRecent progress in microfabrication of biodegradable materials has resulted in the development of a three-dimensional construct suitable for use as a scaffold for engineering blood vessel networks. These networks are designed to replicate the critical fluid dynamic properties of physiological systems such as the microcirculation within a vital organ. Ultimately, these 3D microvascular constructs will serve as a framework for population with organ-specific cells for applications in organ assist and organ replacement. This approach for tissue engineering utilizes highly engineered designs and microfabrication technology to assemble cells in three-dimensional constructs which have physiological values for properties such as mechanical strength, oxygen, nutrient and waste transport, and fluidic parameters such as flow volume and pressure.Three-dimensional networks with appropriate values for blood flow velocity, pressure drop and hematocrit distribution have been designed and fabricated using replica molding techniques, and populated with endothelial cells for long-term microfluidic cell culture. One critical aspect of the fluid dynamics of these systems is the shear stress exerted by blood flow at the walls of the vessel; a key parameter because of well-known mechanotransduction phenomena from mechanical shear forces which govern endothelial cell behavior. In this work, we report the design and construction of three-dimensional microfluidic constructs for tissue engineering which have uniform wall shear stress throughout the network. This type of control over the shear stress offers several advantages over earlier approaches, including more uniform seeding, more rapid achievement of confluent coatings, and better control over endothelial cell behavior for in vitro and in vivo studies.

Intracellular diaphragmed fenestrae in cultured capillary endothelial cells

1988 ◽  
Vol 89 (3) ◽  
pp. 441-447 ◽  
Author(s):  
R. Montesano ◽  
L. Orci
Keyword(s):  
Endothelial Cells ◽  
Endothelial Cell ◽  
Three Dimensional ◽  
Collagen Matrix ◽  
Cell Cytoplasm ◽  
Experimental Conditions ◽  
Intracellular Location ◽  
Capillary Endothelial Cells

The endothelium of visceral capillaries is characterized by the occurrence of numerous fenestrae, which are usually bridged by a thin, single-layered diaphragm. Both in vivo and in vitro, diaphragmed fenestrae perforate the endothelial cell cytoplasm in the most attenuated regions of the cell. We report here that in capillary endothelial cells grown under experimental conditions promoting the development of intracellular lumina (for example, suspension within a three-dimensional collagen matrix), diaphragmed fenestrae can form in a unique, previously undescribed intracellular location - that is, within thin cytoplasmic septa separating contiguous luminal compartments.

scholarly journals Perfusion Flow Enhances Osteogenic Gene Expression and the Infiltration of Osteoblasts and Endothelial Cells into Three-Dimensional Calcium Phosphate Scaffolds

2012 ◽  
Vol 2012 ◽  
pp. 1-10 ◽  
Author(s):  
Matthew J. Barron ◽  
Jeremy Goldman ◽  
Chung-Jui Tsai ◽  
Seth W. Donahue
Keyword(s):  
Gene Expression ◽  
Endothelial Cells ◽  
Endothelial Cell ◽  
Three Dimensional ◽  
Mrna Levels ◽  
Cellular Viability ◽  
Vegf Mrna ◽  
Perfusion Flow

Maintaining cellular viabilityin vivoandin vitrois a critical issue in three-dimensional bone tissue engineering. While the use of osteoblast/endothelial cell cocultures on three-dimensional constructs has shown promise for increasingin vivovascularization,in vitromaintenance of cellular viability remains problematic. This study used perfusion flow to increase osteogenic and angiogenic gene expression, decrease hypoxic gene expression, and increase cell and matrix coverage in osteoblast/endothelial cell co-cultures. Mouse osteoblast-like cells (MC3T3-E1) were cultured alone and in co-culture with mouse microvascular endothelial cells (EOMA) on three-dimensional scaffolds for 1, 2, 7, and 14 days with or without perfusion flow. mRNA levels were determined for several osteogenic, angiogenic, and hypoxia-related genes, and histological analysis was performed. Perfusion flow downregulated hypoxia-related genes (HIF-1α, VEGF, and OPN) at early timepoints, upregulated osteogenic genes (ALP and OCN) at 7 days, and downregulated RUNX-2 and VEGF mRNA at 14 days in osteoblast monocultures. Perfusion flow increased cell number, coverage of the scaffold perimeter, and matrix area in the center of scaffolds at 14 days. Additionally, perfusion flow increased the length of endothelial cell aggregations within co-cultures. These suggest perfusion stimulated co-cultures provide a means of increasing osteogenic and angiogenic activity.

Abstract 565: LPA-PKD-1-HDAC7/NCoR1-FoxO1 Signaling Axis Regulates Endothelial Cell CD36 Transcription and Stimulates Arteriogenic Responses

2014 ◽  
Vol 34 (suppl_1) ◽  
Author(s):  
Bin Ren ◽  
Devi P Ramakrishnan ◽  
Brian Walcott ◽  
Yiliang Chen ◽  
Brad Best ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Endothelial Cell ◽  
Malignant Tumors ◽  
Three Dimensional ◽  
Boyden Chamber ◽  
Dependent Manner ◽  
Novel Approach ◽  
Signaling Axis

Lysophosphatidic acid (LPA), a bioactive signaling phospholipid, down-regulates CD36 expression in microvascular endothelial cells (MVECs) via protein kinase PKD-1 signaling, thereby abolishing endothelial cell responses to its antiangiogenic ligand thrombospondin-1. However, little is known regarding mechanisms by which MVEC-specific CD36 transcription is regulated. We describe that in MVECs LPA represses CD36 transcription by activating a PKD-1 signaling that induces formation of a HDAC7/NCoR1/FoxO1 complex in the nucleus. Promoter analysis first identified FoxO1 as a transcription factor responsible for the CD36 transcription, which was confirmed by a chromatin-immunoprecipitation assay. Using a combination of PKD-1 gene transduction with co-immmunoprecipitation assay, we showed an increased interaction of HDAC7/NCoR1 with FoxO1 in response to LPA. However, HDAC7 and FoxO1 interaction was attenuated with PKD-1 silencing. Furthermore, based on results from an angiogenesis profiling with real time qPCR, doxycycline inducible constitutively active PKD-1 plasmids were transduced into tumor associated endothelial cells using a Lentiviral system to induce the PKD-1 expression. The results showed that turning off CD36 transcription reprograms by PKD-1 signaling was accompanied by an induced expression of ephrin B2 and activation of MAPK/ERK1/2 signaling, which are two critical “molecular signatures” involved in arteriogenesis. Moreover, three dimensional spheroid assay, a modified Boyden Chamber assay and in vivo Matrigel assay revealed that turning off CD36 transcription promoted angiogenesis in vitro and in vivo in a PKD-1-dependent manner. Immunofluorescence microscopy also showed the presence of this signaling pathway in the vasculature of Lewis lung carcinomas grown in cd36 deficient mice. In summary, our data suggest that a LPA-PKD-1-HDAC7/NCoR1-FoxO1 signaling axis is critical for transcriptional regulation of CD36 and mediates silencing of this antiangiogenic switch. This subsequently results in MVEC reprogramming for proangiogenic and arteriogenic responses. Therefore, targeting this signaling cascade could be a novel approach for malignant tumors, cardiovascular ischemia and other thrombotic diseases.

Design and Fabrication of a Constant Shear Microfluidic Network for Tissue Engineering

2004 ◽  
Vol 823 ◽  
Author(s):  
E.J. Weinberg ◽  
J.T. Borenstein ◽  
M.R. Kaazempur-Mofrad ◽  
B. Orrick ◽  
J.P. Vacanti
Keyword(s):  
Tissue Engineering ◽  
Blood Flow ◽  
Shear Stress ◽  
Endothelial Cell ◽  
Dynamic Properties ◽  
Fluid Dynamic ◽  
Three Dimensional ◽  
Cell Behavior

AbstractRecent progress in microfabrication of biodegradable materials has resulted in the development of a three-dimensional construct suitable for use as a scaffold for engineering blood vessel networks. These networks are designed to replicate the critical fluid dynamic properties of physiological systems such as the microcirculation within a vital organ. Ultimately, these 3D microvascular constructs will serve as a framework for population with organ-specific cells for applications in organ assist and organ replacement. This approach for tissue engineering utilizes highly engineered designs and microfabrication technology to assemble cells in three-dimensional constructs which have physiological values for properties such as mechanical strength, oxygen, nutrient and waste transport, and fluidic parameters such as flow volume and pressure.Three-dimensional networks with appropriate values for blood flow velocity, pressure drop and hematocrit distribution have been designed and fabricated using replica molding techniques, and populated with endothelial cells for long-term microfluidic cell culture. One critical aspect of the fluid dynamics of these systems is the shear stress exerted by blood flow at the walls of the vessel; a key parameter because of well-known mechanotransduction phenomena from mechanical shear forces which govern endothelial cell behavior. In this work, we report the design and construction of three-dimensional microfluidic constructs for tissue engineering which have uniform wall shear stress throughout the network. This type of control over the shear stress offers several advantages over earlier approaches, including more uniform seeding, more rapid achievement of confluent coatings, and better control over endothelial cell behavior for in vitro and in vivo studies.

In vitro and in vivo polysaccharides assembly: ultrastructural and cytochemical study of growing plant cell wall components.

1978 ◽  
Vol 36 (2) ◽  
pp. 434-435
Author(s):  
D. Reis ◽  
B. Vian ◽  
J. C. Roland
Keyword(s):  
Cell Wall ◽  
Self Assembly ◽  
Plant Cell Wall ◽  
Higher Plants ◽  
Three Dimensional ◽  
Cytochemical Study ◽  
Wall Components

Wall morphogenesis in higher plants is a problem still open to controversy. Until now the possibility of a transmembrane control and the involvement of microtubules were mostly envisaged. Self-assembly processes have been observed in the case of walls of Chlamydomonas and bacteria. Spontaneous gelling interactions between xanthan and galactomannan from Ceratonia have been analyzed very recently. The present work provides indications that some processes of spontaneous aggregation could occur in higher plants during the formation and expansion of cell wall.Observations were performed on hypocotyl of mung bean (Phaseolus aureus) for which growth characteristics and wall composition have been previously defined.In situ, the walls of actively growing cells (primary walls) show an ordered three-dimensional organization (fig. 1). The wall is typically polylamellate with multifibrillar layers alternately transverse and longitudinal. Between these layers intermediate strata exist in which the orientation of microfibrils progressively rotates. Thus a progressive change in the morphogenetic activity occurs.

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