scholarly journals Moderate Hypothermia Has the Potential to Reveal the Dominant/Submissive Relationship in a Co-Culture System Consisting of Osteoblasts and Endothelial Cells

Micro ◽  
2021 ◽  
Vol 1 (2) ◽  
pp. 181-193
Author(s):  
Kouki Inomata ◽  
Michiyo Honda
Keyword(s):  
Tissue Engineering ◽  
Endothelial Cells ◽  
Culture System ◽  
Spatial Organization ◽  
Cellular Proliferation ◽  
Cell Types ◽  
Culture Conditions ◽  
Moderate Hypothermia ◽  
Bone Homeostasis ◽  
And Migration

Microvessels in bone are indispensable for maintaining bone homeostasis based on a dynamic remodeling system. In cell-based tissue engineering, vascularization into the regenerative bone is a key strategy to avoid hypoxia and necrosis around re-implanted tissues. Previous studies have shown that direct contact between osteoblasts and endothelial cells stimulates differentiation of both cell types. However, no studies have revealed the dominant/submissive relationship. In the present study, we examined the effect of hypothermia on monoculture and co-culture to assess which cells tightly coordinated osteogenesis and angiogenesis in the co-culture system. As for osteoblasts, exposure to hypothermia suppressed cellular proliferation, migration, and differentiation. Evaluation of the behavior of endothelial cells showed that hypothermia should not affect basic functions such as proliferation and migration. Under co-culture conditions, both osteogenic differentiation and the formation of vessel-like angiogenic structures were suppressed by hypothermia, but the spatial organization of alkaline phosphatase-positive cell clusters, which tend to localize around microvascular lumens, was not altered. These data suggest that hypothermia attenuates heterotypic intercellular crosstalk which robustly depends on osteoblasts to inhibit both osteogenesis and angiogenesis in the co-culture system. Taken together, this approach will provide new insights into the relationship between osteoblasts and endothelial cells in tissue engineering.

scholarly journals BIM deficiency differentially impacts the function of kidney endothelial and epithelial cells through modulation of their local microenvironment

2012 ◽  
Vol 302 (7) ◽  
pp. F809-F819 ◽  
Author(s):  
Nader Sheibani ◽  
Margaret E. Morrison ◽  
Zafer Gurel ◽  
SunYoung Park ◽  
Christine M. Sorenson
Keyword(s):  
Nitric Oxide ◽  
Endothelial Cells ◽  
Epithelial Cells ◽  
Cell Types ◽  
Proper Function ◽  
Causative Role ◽  
Structural Support ◽  
Cell Adhesion And Migration ◽  
And Migration ◽  
Endothelial Nitric Oxide

The extracellular matrix (ECM) acts as a scaffold for kidney cellular organization. Local secretion of the ECM allows kidney cells to readily adapt to changes occurring within the kidney. In addition to providing structural support for cells, the ECM also modulates cell survival, migration, proliferation, and differentiation. Although aberrant regulation of ECM proteins can play a causative role in many diseases, it is not known whether ECM production, cell adhesion, and migration are regulated in a similar manner in kidney epithelial and endothelial cells. Here, we demonstrate that lack of BIM expression differentially impacts kidney endothelial and epithelial cell ECM production, migration, and adhesion, further emphasizing the specialized role of these cell types in kidney function . Bim −/− kidney epithelial cells demonstrated decreased migration, increased adhesion, and sustained expression of osteopontin and thrombospondin-1 (TSP1). In contrast, bim −/− kidney endothelial cells demonstrated increased cell migration, and decreased expression of osteopontin and TSP1. We also observed a fivefold increase in VEGF expression in bim −/− kidney endothelial cells consistent with their increased migration and capillary morphogenesis. These cells also had decreased endothelial nitric oxide synthase activity and nitric oxide bioavailability. Thus kidney endothelial and epithelial cells make unique contributions to the regulation of their ECM composition, with specific impact on adhesive and migratory properties that are essential for their proper function.

scholarly journals Capillary-Like Formations of Endothelial Cells in Defined Patterns Generated by Laser Bioprinting

Micromachines ◽  
2021 ◽  
Vol 12 (12) ◽  
pp. 1538
Author(s):  
Lothar Koch ◽  
Andrea Deiwick ◽  
Boris Chichkov
Keyword(s):  
Tissue Engineering ◽  
Endothelial Cells ◽  
High Cell Density ◽  
Cell Types ◽  
High Cell ◽  
Tubular Structures ◽  
Promising Technique ◽  
External Stimulation ◽  
Random Patterns ◽  
Printed Patterns

Bioprinting is seen as a promising technique for tissue engineering, with hopes of one day being able to produce whole organs. However, thick tissue requires a functional vascular network, which naturally contains vessels of various sizes, down to capillaries of ~10 µm in diameter, often spaced less than 200 µm apart. If such thick tissues are to be printed, the vasculature would likely need to be printed at the same time, including the capillaries. While there are many approaches in tissue engineering to produce larger vessels in a defined manner, the small capillaries usually arise only in random patterns by sprouting from the larger vessels or from randomly distributed endothelial cells. Here, we investigated whether the small capillaries could also be printed in predefined patterns. For this purpose, we used a laser-based bioprinting technique that allows for the combination of high resolution and high cell density. Our aim was to achieve the formation of closed tubular structures with lumina by laser-printed endothelial cells along the printed patterns on a surface and in bioprinted tissue. This study shows that such capillaries are directly printable; however, persistence of the printed tubular structures was achieved only in tissue with external stimulation by other cell types.

scholarly journals Chitlac-coated Thermosets Enhance Osteogenesis and Angiogenesis in a Co-culture of Dental Pulp Stem Cells and Endothelial Cells

Nanomaterials ◽  
2019 ◽  
Vol 9 (7) ◽  
pp. 928 ◽  
Author(s):  
Rapino ◽  
Di Valerio ◽  
Zara ◽  
Gallorini ◽  
Marconi ◽  
...  
Keyword(s):  
Stem Cells ◽  
Endothelial Cells ◽  
Dental Pulp ◽  
Cellular Response ◽  
Cell Types ◽  
Culture Conditions ◽  
Dental Pulp Stem Cells ◽  
Alizarin Red ◽  
Dental Restorations ◽  
Diphenyl Tetrazolium Bromide

Dental pulp stem cells (DPSCs) represent a population of stem cells which could be useful in oral and maxillofacial reconstruction. They are part of the periendothelial niche, where their crosstalk with endothelial cells is crucial in the cellular response to biomaterials used for dental restorations. DPSCs and the endothelial cell line EA.hy926 were co-cultured in the presence of Chitlac-coated thermosets in culture conditions inducing, in turn, osteogenic or angiogenic differentiation. Cell proliferation was evaluated by 3–[4,5–dimethyl–thiazol–2–yl–]–2,5–diphenyl tetrazolium bromide (MTT) assay. DPSC differentiation was assessed by measuring Alkaline Phosphtase (ALP) activity and Alizarin Red S staining, while the formation of new vessels was monitored by optical microscopy. The IL-6 and PGE2 production was evaluated as well. When cultured together, the proliferation is increased, as is the DPSC osteogenic differentiation and EA.hy926 vessel formation. The presence of thermosets appears either not to disturb the system balance or even to improve the osteogenic and angiogenic differentiation. Chitlac-coated thermosets confirm their biocompatibility in the present co-culture model, being capable of improving the differentiation of both cell types. Furthermore, the assessed co-culture appears to be a useful tool to investigate cell response toward newly synthesized or commercially available biomaterials, as well as to evaluate their engraftment potential in restorative dentistry.

The regulation of E3 ubiquitin ligases Cbl and its cross talking in bone homeostasis

2020 ◽  
Vol 15 ◽  
Author(s):  
Xiaobin Yang ◽  
Dingjun Hao ◽  
Baorong He
Keyword(s):  
Cellular Proliferation ◽  
Bone Matrix ◽  
Osteoclast Differentiation ◽  
Ubiquitin Ligases ◽  
E3 Ubiquitin Ligases ◽  
Bone Homeostasis ◽  
And Migration ◽  
First Time ◽  
Definite Role

: The E3 ubiquitin ligases Cbl has been found play an important role in regulating cellular proliferation and migration. Whereas the excessive differentiation of osteoclast and/or its over expressing of resorptive functions could lead the pathological bone homeostasis by overly bone matrix degradation. Since the first time of the important role of Cbl in the regulating osteoclast differentiation (also named osteoclastogenesis) has been reported in decades ago. The extensively studies have been conducted for in-depth exploring the Cbl’s definite role during osteoclastogenesis, as well as its cross talking with other signaling pathways (such as: Src and PI3K signaling) in bone homeostasis. Herein, our current study aim to briefly conclude the current studies of osteoclastogenesis and the regulatory role of Cbl, as well as its cross-talking in bone homeostasis.

Evidence of synergistic/additive effects of sildenafil and erythropoietin in enhancing survival and migration of hypoxic endothelial cells

2013 ◽  
Vol 304 (4) ◽  
pp. L230-L239 ◽  
Author(s):  
Elena Gammella ◽  
Caroline Leuenberger ◽  
Max Gassmann ◽  
Louise Ostergaard
Keyword(s):  
Endothelial Cells ◽  
Combination Treatment ◽  
Synergistic Effects ◽  
Annexin V ◽  
Cell Types ◽  
Blue Staining ◽  
Protective Effects ◽  
Hypoxic Conditions ◽  
Phosphodiesterase Type 5 Inhibitor ◽  
And Migration

Endothelial cell dysfunction is a common event to several pathologies including pulmonary hypertension, which is often associated with hypoxia. As the endothelium plays an essential role in regulating the dynamic interaction between pulmonary vasodilatation and vasoconstriction, this cell type is fundamental in the development of vascular remodeling and increased vascular resistance. We investigated the protective effects of sildenafil, a phosphodiesterase type 5 inhibitor, given in combination with erythropoietin (Epo), as it has been demonstrated that both drugs have antiapoptotic effects on several cell types. Specifically, we examined the viability and angiogenic properties of rat pulmonary artery endothelial cells upon exposure to either 21% or 1% oxygen, in presence of sildenafil (1 and 100 nM) and Epo (5 and 20 U/ml) alone or in combination (1 nM and 20 U/ml). Cell proliferation and viability were analyzed by Trypan blue staining, MTT assay, and Annexin V/propidium iodide stainings. In all assays, the ability of the combination treatment in improving cell viability was superior to that of either drug alone. The angiogenic properties were studied using a migration and a 3D collagen assay, and the results revealed increases in the migration potential of endothelial cells as well as the ability to form tube-like structures in response to sildenafil and the combination treatment. We therefore conclude that both drugs exert protective effects on endothelial cells on hypoxia and that sildenafil enhances the migratory and angiogenic properties, especially in hypoxic conditions. Furthermore, we present evidence of possible additive or synergistic effects of both drugs.

Effects of culture conditions on the proliferation, morphology and migration of bovine aortic endothelial cells

1983 ◽  
Vol 62 (1) ◽  
pp. 267-285
Author(s):  
A.M. Schor ◽  
S.L. Schor ◽  
T.D. Allen
Keyword(s):  
Endothelial Cells ◽  
Three Dimensional ◽  
Collagen Gel ◽  
Culture Conditions ◽  
Apical Surface ◽  
Collagen Gels ◽  
Aortic Endothelial Cells ◽  
Plating Density ◽  
Adhesive Interactions ◽  
And Migration

Various culture conditions, such as the presence of ascorbic acid, initial plating density and the nature of the substratum (plastic, gelatin or native collagen gels), influenced the growth, morphology and migration of three cloned populations of adult bovine aorta endothelial cells. Aorta endothelial cells showed two distinctive and reversible morphological phenotypes. Cells presenting a free apical surface were polygonal and formed sheets of overlapping or non-overlapping cells, depending on the culture conditions. When the cells were able to establish adhesive interactions over their entire cell surface they adopted an elongated shape and formed meshworks of interconnected ‘sprouting’ cells. The cells were capable of migrating into a collagen gel from both their basal and apical surfaces. Once in the gel, they formed characteristic, compact, three-dimensional meshworks.

Adenosine and hypoxia stimulate proliferation and migration of endothelial cells

1988 ◽  
Vol 255 (3) ◽  
pp. H554-H562 ◽  
Author(s):  
C. J. Meininger ◽  
M. E. Schelling ◽  
H. J. Granger
Keyword(s):  
Cell Proliferation ◽  
Endothelial Cells ◽  
Culture Medium ◽  
Culture Conditions ◽  
Hypoxic Conditions ◽  
Cell Counts ◽  
And Migration ◽  
Stimulate Cell Proliferation ◽  
Proliferation And Migration

The proliferation of bovine aortic or coronary venular endothelial cells (EC) in vitro was stimulated by the addition of adenosine (0.5 or 5.0 microM) to the culture medium. Cell counts of adenosine-treated aortic EC were 23–76% and coronary venular EC 19–52% greater than nontreated controls. Because adenosine is known to be released by hypoxic tissues, cell proliferation was quantitated when aortic EC were grown at 2% O2. Cell counts were 41–102% greater under hypoxic conditions than when cells were grown at standard tissue culture conditions (approximately 20% O2). When culture medium conditioned by coronary EC grown at 2% O2 was added to EC growing at standard conditions, cell counts were 24–69% greater than controls with medium conditioned by coronary EC grown at 20% O2. This suggests that hypoxia causes endothelial cells to release a factor(s) into the medium that can stimulate cell proliferation. The addition of the adenosine receptor blocker 8-phenyltheophylline (10(-5) M) prevented the stimulation of proliferation caused by hypoxia-conditioned medium, 2% O2 or 5.0 microM adenosine, suggesting that adenosine mediates its effect via an external membrane receptor. Adenosine also stimulated EC chemotaxis. Taken together, these results suggest that adenosine, released as a result of tissue hypoxia, may act as an angiogenic stimulus for the growth of new blood vessels.

scholarly journals Small organelle, big responsibility: the role of centrosomes in development and disease

2014 ◽  
Vol 369 (1650) ◽  
pp. 20130468 ◽  
Author(s):  
Pavithra L. Chavali ◽  
Monika Pütz ◽  
Fanni Gergely
Keyword(s):  
Developmental Disorders ◽  
Primary Cilia ◽  
Spatial Organization ◽  
Growth Failure ◽  
Cell Types ◽  
Model Systems ◽  
And Migration ◽  
Division Cell

The centrosome, a key microtubule organizing centre, is composed of centrioles, embedded in a protein-rich matrix. Centrosomes control the internal spatial organization of somatic cells, and as such contribute to cell division, cell polarity and migration. Upon exiting the cell cycle, most cell types in the human body convert their centrioles into basal bodies, which drive the assembly of primary cilia, involved in sensing and signal transduction at the cell surface. Centrosomal genes are targeted by mutations in numerous human developmental disorders, ranging from diseases exclusively affecting brain development, through global growth failure syndromes to diverse pathologies associated with ciliary malfunction. Despite our much-improved understanding of centrosome function in cellular processes, we know remarkably little of its role in the organismal context, especially in mammals. In this review, we examine how centrosome dysfunction impacts on complex physiological processes and speculate on the challenges we face when applying knowledge generated from in vitro and in vivo model systems to human development.

Tissue Engineering: Controlling Spatial Organization of Multiple Cell Types in Defined 3D Geometries (Adv. Mater. 41/2012)

2012 ◽  
Vol 24 (41) ◽  
pp. 5542-5542
Author(s):  
Halil Tekin ◽  
Jefferson G. Sanchez ◽  
Christian Landeros ◽  
Karen Dubbin ◽  
Robert Langer ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Spatial Organization ◽  
Cell Types ◽  
Multiple Cell

scholarly journals Bioprinting with human stem cells-laden alginate-gelatin bioink and bioactive glass for tissue engineering

2019 ◽  
Vol 5 (2.2) ◽  
pp. 3 ◽  
Author(s):  
Krishna C. R. Kolan ◽  
Julie A. Semon ◽  
Bradley Bromet ◽  
Delbert E. Day ◽  
Ming C. Leu
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Bioactive Glass ◽  
Three Dimensional ◽  
Cell Types ◽  
3D Models ◽  
Culture Conditions ◽  
3D Bioprinting ◽  
Human Stem Cells ◽  
Tissue Repair And Regeneration

Three-dimensional (3D) bioprinting technologies have shown great potential in the fabrication of 3D models for different human tissues. Stem cells are an attractive cell source in tissue engineering as they can be directed by material and environmental cues to differentiate into multiple cell types for tissue repair and regeneration. In this study, we investigate the viability of human adipose-derived mesenchymal stem cells (ASCs) in alginate-gelatin (Alg-Gel) hydrogel bioprinted with or without bioactive glass. Highly angiogenic borate bioactive glass (13-93B3) in 50 wt% is added to polycaprolactone (PCL) to fabricate scaffolds using a solvent-based extrusion 3D bioprinting technique. The fabricated scaffolds with 12 × 12 × 1 mm3 in overall dimensions are physically characterized, and the glass dissolution from PCL/glass composite over a period of 28 days is studied. Alg-Gel composite hydrogel is used as a bioink to suspend ASCs, and scaffolds are then bioprinted in different configurations: Bioink only, PCL+bioink, and PCL/glass+bioink, to investigate ASC viability. The results indicate the feasibility of the solvent-based bioprinting process to fabricate 3D cellularized scaffolds with more than 80% viability on day 0. The decrease in viability after 7 days due to glass concentration and static culture conditions is discussed. The feasibility of modifying Alg-Gel with 13-93B3 glass for bioprinting is also investigated, and the results are discussed.

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