scholarly journals Photocrosslinkable polysaccharide hydrogel composites based on dextran or pullulan–amylose blends with cytokines for a human co-culture model of human osteoblasts and endothelial cells

2016 ◽  
Vol 4 (40) ◽  
pp. 6552-6564 ◽  
Author(s):  
Ulrike Ritz ◽  
Peter Kögler ◽  
Isabel Höfer ◽  
Petra Frank ◽  
Sven Klees ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Bone Regeneration ◽  
Human Osteoblasts ◽  
Culture Model ◽  
Hydrogel Composites ◽  
Polysaccharide Hydrogel

Polysaccharide hyrogel composites demonstrate fundamental potential as biomaterials for bone regeneration in vitro.

scholarly journals Modulation of Wnt/β-catenin signaling promotes blood-brain barrier phenotype in cultured brain endothelial cells

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Marlyn D. Laksitorini ◽  
Vinith Yathindranath ◽  
Wei Xiong ◽  
Sabine Hombach-Klonisch ◽  
Donald W. Miller
Keyword(s):  
Endothelial Cells ◽  
Wnt Signaling ◽  
Blood Brain Barrier ◽  
Brain Barrier ◽  
Brain Endothelial Cells ◽  
Cancer Resistance ◽  
Blood Brain ◽  
Culture Model ◽  
External Activation

AbstractWnt/β-catenin signaling is important for blood-brain barrier (BBB) development and is implicated in BBB breakdown under various pathophysiological conditions. In the present study, a comprehensive characterization of the relevant genes, transport and permeability processes influenced by both the autocrine and external activation of Wnt signaling in human brain endothelial cells was examined using hCMEC/D3 culture model. The hCMEC/D3 expressed a full complement of Wnt ligands and receptors. Preventing Wnt ligand release from hCMEC/D3 produced minimal changes in brain endothelial function, while inhibition of intrinsic/autocrine Wnt/β-catenin activity through blocking β-catenin binding to Wnt transcription factor caused more modest changes. In contrast, activation of Wnt signaling using exogenous Wnt ligand (Wnt3a) or LiCl (GSK3 inhibitor) improved the BBB phenotypes of the hCMEC/D3 culture model, resulting in reduced paracellular permeability, and increased P-glycoprotein (P-gp) and breast cancer resistance associated protein (BCRP) efflux transporter activity. Further, Wnt3a reduced plasmalemma vesicle associated protein (PLVAP) and vesicular transport activity in hCMEC/D3. Our data suggest that this in vitro model of the BBB has a more robust response to exogenous activation of Wnt/β-catenin signaling compared to autocrine activation, suggesting that BBB regulation may be more dependent on external activation of Wnt signaling within the brain microvasculature.

A Novel Bone Scaffolds Based on Hyperbranched Polyglycerol Fibers Filled with Hydroxyapatite Nanoparticles: In Vitro Cell Response

2008 ◽  
Vol 396-398 ◽  
pp. 633-636 ◽  
Author(s):  
Alvaro Antonio Alencar de Queiroz ◽  
José Carlos Bressiani ◽  
Ana Helena A. Bressiani ◽  
Olga Z. Higa ◽  
Gustavo Abel Abraham
Keyword(s):  
Alkaline Phosphatase ◽  
Bone Regeneration ◽  
Cell Response ◽  
Hydroxyapatite Nanoparticles ◽  
Human Osteoblasts ◽  
Bone Scaffolds ◽  
Solution Cast ◽  
Hyperbranched Polyglycerol ◽  
Potential Use

A novel bone scaffolding material was successfully fabricated by electrospinning from hyperbranched polyglycerol (HPGL) solutions containing nanoparticles of hydroxyapatite (HA). The potential use of the electrospun fibrous HPGL-HA scaffolds for bone regeneration was evaluated in vitro with human osteoblasts in terms of alkaline phosphatase (ALP) activity of the cells that were cultured directly on the scaffolds. The results were compared with those on corresponding HPLG-HA solution-cast film scaffolds. It was found that all of the fibrous scaffolds promoted much better adhesion and proliferation of cells than the corresponding film scaffolds.

Fabrication and in vitro biological evaluation of photopolymerisable hydroxyapatite hydrogel composites for bone regeneration

2013 ◽  
Vol 28 (8) ◽  
pp. 1274-1283 ◽  
Author(s):  
John A Killion ◽  
Luke M Geever ◽  
Declan M Devine ◽  
Clement L Higginbotham
Keyword(s):  
Bone Regeneration ◽  
Biological Evaluation ◽  
Hydrogel Composites

Observations on Biological Effects of Carbon-13, Based on the Effects of 13C-Testosterone on Growth of Human Osteoblasts, Human Aortic Endothelial Cells, and Human Umbilical Vein Endothelial Cells in Vitro

2021 ◽  
Author(s):  
Mengqi Zhang ◽  
Wenning Yang ◽  
Xinchen Wu ◽  
Tengfei Zhang
Keyword(s):  
Cell Proliferation ◽  
Endothelial Cells ◽  
Isotope Effect ◽  
Umbilical Vein ◽  
Biological Effects ◽  
Bioactive Compound ◽  
Human Osteoblasts ◽  
Aortic Endothelial Cells ◽  
Umbilical Vein Endothelial Cells

Abstract Despite the increasing knowledge of biological isotope effect, comprehensive understanding of heavy isotope effect in the biological contexts has remained far less than expectation. The present study investigated the carbon isotope effect of 13C enriched testosterone on human cells. It was among the rare studies on carbon isotope effect of bioactive compound. Human osteoblasts, human aortic endothelial cells, and human umbilical vein endothelial cells were cultured in vitro and treated with testosterone and 13C enriched testosterone (13C/12C:6.7%). The impacts of physiological to pharmacological concentrations (10-10-10-5mol/L) of the bioactive compound were taken into account. The cell proliferation activities were measured using MTS assay. The levels of alkaline phosphatase and osteocalcin in osteoblasts were tested. Our results established that 13C enriched testosterone exhibited different biological effects from testosterone. At the concentrations of 10-10mol/L and 10-5mol/L, there were significant differences in prompting cell proliferation between testosterone and 13C enriched testosterone. At physiological concentrations, testosterone prompted proliferations of the three kinds of cells; whereas, 13C enriched testosterone did not prompt the cell proliferation, and its effects were not concentration dependent. At supraphysiological concentration (10-5mol/L), testosterone had the trend of inhibiting cell growth; whereas, 13C enriched testosterone had the trend of prompting cell growth. 13C enriched testosterone significantly enhanced osteocalcin secretion in human osteoblasts at supraphysiological concentration. These findings challenged the common view of growth retardation effect of heavy isotope, which imply that biological isotope effects are worthy of further study. The potential applications of 13C enriched compound were discussed.

scholarly journals Establishment of murine in vitro blood-brain barrier models using immortalized cell lines: co-cultures of brain endothelial cells, astrocytes, and neurons

2018 ◽  
Author(s):  
Fakhriedzwan Idris ◽  
Siti Hanna Muharram ◽  
Zainun Zaini ◽  
Suwarni Diah
Keyword(s):  
Endothelial Cells ◽  
Blood Brain Barrier ◽  
Cell Lines ◽  
Brain Barrier ◽  
Brain Endothelial Cells ◽  
Immortalized Cell Lines ◽  
Culture Model ◽  
Immortalized Cell

AbstractBlood-brain barrier (BBB) is a selective barrier formed by the endothelial cells that line cerebral microvessels. It serves as a physical barrier due to the presence of complex tight junctions between adjacent endothelial cells which limits the paracellular movement of most molecules across the BBB. Many in vitro models of the BBB have been established to mimic these in vivo properties with limited success. In this study, we described the properties of a cell-based murine in vitro BBB model in five configurations constructed using immortalized cell lines in a 12-well format Transwell system: murine brain endothelial cells (bEnd.3) grown in a monoculture, or as co-culture in contact with astrocytes, or without contact with astrocytes or neurons, and triple co-culture combining the three cell lines. We found that only contact and triple co-culture model closely mimic the in vivo BBB tightness as evaluated by apparent permeability (Papp) of sucrose and albumin producing the lowest Papp values of 0.56 ± 0.16 × 10−6 cms−1 and 3.30 ± 0.51 × 10−6 cms−1, respectively, obtained in triple co-culture model. Co-culturing of bEnd.3 with astrocytes increased the expression of occludin as shown by western blot analysis, and immunohistochemistry showed an increase in peripheral localization of occludin and claudin-5. In addition, we found conditioned media were able to increase in vitro BBB model tightness through the modulation of tight junction proteins localization. We conclude that the presence of astrocytes and neurons in close proximity to brain endothelial cells is essential to produce a tight in vitro BBB model.

scholarly journals Triple Culture of Primary Human Osteoblasts, Osteoclasts and Osteocytes as an In Vitro Bone Model

2021 ◽  
Vol 22 (14) ◽  
pp. 7316
Author(s):  
Anne Bernhardt ◽  
Jasmin Skottke ◽  
Max von von Witzleben ◽  
Michael Gelinsky
Keyword(s):  
Final Analysis ◽  
Cell Types ◽  
Porous Membrane ◽  
Collagen Gels ◽  
Human Osteoblasts ◽  
Culture Model ◽  
Typical Morphology ◽  
Primary Human Osteoblasts ◽  
Different Cell Types

In vitro evaluation of bone graft materials is generally performed by analyzing the interaction with osteoblasts or osteoblast precursors. In vitro bone models comprising different cell species can give specific first information on the performance of those materials. In the present study, a 3D co-culture model was established comprising primary human osteoblasts, osteoclasts and osteocytes. Osteocytes were differentiated from osteoblasts embedded in collagen gels and were cultivated with osteoblast and osteoclasts seeded in patterns on a porous membrane. This experimental setup allowed paracrine signaling as well as separation of the different cell types for final analysis. After 7 days of co-culture, the three cell species showed their typical morphology and gene expression of typical markers like ALPL, BSPII, BLGAP, E11, PHEX, MEPE, RANKL, ACP5, CAII and CTSK. Furthermore, relevant enzyme activities for osteoblasts (ALP) and osteoclasts (TRAP, CTSK, CAII) were detected. Osteoclasts in triple culture showed downregulated TRAP (ACP5) and CAII expression and decreased TRAP activity. ALP and BSPII expression of osteoblasts in triple culture were upregulated. The expression of the osteocyte marker E11 (PDPN) was unchanged; however, osteocalcin (BGLAP) expression was considerably downregulated both in osteoblasts and osteocytes in triple cultures compared to the respective single cultures.

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