The Effects of Hyperosmolarity on the Viability and Function of Endothelial Cells

Luh, E.H.; Shackford, Steven R.; Shatos, Marie A.; Pietropaoli, John A.

doi:10.1006/jsre.1996.0020

Cited by 19 publications

(10 citation statements)

References 11 publications

(14 reference statements)

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“…To avoid unspecific cytotoxic effects, all polymer solutions were adjusted to physiological pH and tolerable osmolarity. Although an osmolarity above 300 mosmol · kg −1 is considered as cytotoxic, our preliminary experiments showed that osmolarities up to 400 mosmol · kg −1 were well tolerated by L929 cells, comparable with literature data on the investigation of bovine aortic endothelium cells 27…”

Section: Discussionsupporting

confidence: 90%

Poly(2‐ethyl‐2‐oxazoline) as Alternative for the Stealth Polymer Poly(ethylene glycol): Comparison of in vitro Cytotoxicity and Hemocompatibility

Bauer

Lautenschlaeger

Kempe

et al. 2012

Macromolecular Bioscience

259

199

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Limitations of PEG in drug delivery have been reported from clinical trials. PEtOx (0.4-40 kDa) as alternative is synthesized by a living, microwave-assisted polymerization, and is directly compared to PEG of similar molar mass regarding cytotoxicity and hemocompatibility. In short-term treatments, both types of polymers are well tolerated even at high concentrations. Moderate concentration and molar mass dependent cytotoxic effects occurred only after long-term incubation at concentrations higher than therapeutic doses. PEtOx possesses not only an easy route of synthesis and beneficial physicochemical characteristics such as low viscosity and high stability, which are advantageous over PEG, but additionally in vitro toxicology comparable to PEG.

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Section: Discussionsupporting

confidence: 90%

Poly(2‐ethyl‐2‐oxazoline) as Alternative for the Stealth Polymer Poly(ethylene glycol): Comparison of in vitro Cytotoxicity and Hemocompatibility

Bauer

Lautenschlaeger

Kempe

et al. 2012

Macromolecular Bioscience

259

199

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“…But in combination with more sensitive and reliable methods such as measurement of LDH release, the trypan blue exclusion test is generally ac cepted. Several authors reported that both methods equal ly well define endothelial cell viability [21,36], H2O2 con centrations above 1 mM showed contraction and round ing of endothelial cells [7]. Using such high concentrations of H2O2 has many times been reported to be related to subsequent toxicity [37], These findings can be inter preted as early morphological phenomena of reactive oxy gen species-induced cell death.…”

Section: Discussionmentioning

confidence: 83%

“…Both exposure to 0% Oi and 3% CT failed to diminish cell viability and to induce endothelial cell swelling. Accord ingly, we believe that in vitro exposure to hypoxia at the levels used led to alterations in the metabolic properties of endothelial cells, in such a way that their structure and vital functions were maintained [21,25], The mecha nisms, however, by which endothelial cells preserve their cellular function and maintain their structure during de creases in ambient 03 tension are not known.…”

Section: Discussionmentioning

confidence: 99%

“…Thus exposure to hypertonic media may lead to no further decrease in cell volume as seen in our experiments using flow cytometry. It was demonstrated by others that vascular endothelial cells tolerate media osmolarities of up to 460 mosm/1 without apparent decrement in viability even in the adverse condition of anoxia in vitro [21]. The aim of the cited study was to evaluate the hyperosmotic tolerance of endothelial cells, because hypertonic solutions have been demonstrated to be efficacious in the treatment of hypo volemic shock [22], The maintenance of functional and morphological integrity of the vascular endothelium is one of the essential aspects of stabilization of microhemo dynamics after trauma or hypovolemic shock.…”

Section: Discussionmentioning

confidence: 99%

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The Influence of Cellular Hypoxia and Reactive Oxygen Species on the Development of Endothelial Cell Edema

Hensel

Volk²,

Kox³

1997

Int J Microcirc

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We investigated in vitro whether endothelial cell edema is induced by cellular hypoxia or oxygen radical formation. Measurements of relative cell volume (RCV) were made using microweight analysis, liquid scintillation spectrometry and analysis of cellular protein content. To validate this method of determining cell volume, endothelial cells were incubated in media of different osmolarities. Vascular endothelial cells reacted to osmotic stress with a volume increase or decrease. The addition of xanthine oxidase (XOD; 3 mU/ml) and hypoxanthine (1 mM) for the enzymatic production of O^-₂ caused a reproducible and significant increase in RCV by 29 ± 8% (from 5.5 to 7.1 µl/10⁶ cells; p < 0.001) after an incubation time of 60 min. Nonenzymatically produced H₂O₂ (100 µM) caused a similar increase in RCV by 35 ± 5% (from 5.5 to 7.6 µl/10⁶ cells; p < 0.001) over the same incubation period. The addition of catalase (50 U/ml) diminished the increasing effect of XOD as well as that of H₂O₂ on cell volume. As assessed by the uptake of the vital dye trypan blue and the release of lactate dehydrogenase into the medium, there was no significant loss of viability during the incubation time. Lower concentrations of H₂O₂ as well as lower activities of XOD did not induce a significant increase in RCV. Higher H₂O₂ concentrations and increased XOD activities caused a considerable time- and concentration-dependent injury of endothelial cells. RCV was unchanged even after long exposure (5 h) to two different hypoxic gas mixtures (3% O₂:5% CO₂:92% N₂;0% O₂:5% CO₂:95%N₂). Cell viability was not impaired under hypoxic conditions. The results suggest that reactive oxygen species play a more important role in the development of endothelial cell edema than cellular hypoxia.

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“…In general, the longevity and functional availability of cell cultures are mainly limited by contamination with pathogens, non-optimal medium composition, mechanical stress as well as fl uctuations in temperature, pH or osmolarity leading to dedifferentiation, senescence, or apoptosis (Kallos et al 1999;Bruner 2003). While hyperosmolarity alters cell physiology and gene expression (Chen et al 2007;Morita 2007;Wuertz et al 2007), it does not seem to limit cell viability per se (Arimochi and Morita 2005;Olsen et al 2005), and does not affect every cell type to the same extent (Luh et al 1996). However, studies have shown that stabilization of osmolarity has a benefi cial effect on the longevity of neurons in cell culture (Maher and McKinney 1995;Potter and Demarse 2001).…”

Section: Introductionmentioning

confidence: 96%

Replica-moulded polydimethylsiloxane culture vessel lids attenuate osmotic drift in long-term cell cultures

et al. 2009

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An imbalance in medium osmolarity is a determinant that affects cell culture longevity. Even in humidified incubators, evaporation of water leads to a gradual increase in osmolarity over time. We present a simple replica-moulding strategy for producing self-sealing lids adaptable to standard, small-size cell-culture vessels. They are made of polydimethylsiloxane (PDMS), a flexible, transparent and biocompatible material, which is gas-permeable but largely impermeable to water. Keeping cell cultures in a humidified 5% CO2 incubator at 37 degrees C, medium osmolarity increased by +6.86 mosmol/kg/day in standard 35 mm Petri dishes, while PDMS lids attenuated its rise by a factor of four to changes of +1.72 mosmol/kg/ day. Depending on the lid membrane thickness,pH drifts at ambient CO2 levels were attenuated by a factor of 4 to 9. Comparative evaporation studies at temperatures below 60 degrees C yielded a 10-fold reduced water vapour flux of 1.75 g/day/ dm 2 through PDMS lids as compared with 18.69 g/day/dm 2 with conventional Petri dishes. Using such PDMS lids,about 2/3 of the cell cultures grew longer than 30 days in vitro. Among these,the average survival time was 69 days with the longest survival being 284 days under otherwise conventional cell culture conditions.

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The Effects of Hyperosmolarity on the Viability and Function of Endothelial Cells

Cited by 19 publications

References 11 publications

Poly(2‐ethyl‐2‐oxazoline) as Alternative for the Stealth Polymer Poly(ethylene glycol): Comparison of in vitro Cytotoxicity and Hemocompatibility

Poly(2‐ethyl‐2‐oxazoline) as Alternative for the Stealth Polymer Poly(ethylene glycol): Comparison of in vitro Cytotoxicity and Hemocompatibility

The Influence of Cellular Hypoxia and Reactive Oxygen Species on the Development of Endothelial Cell Edema

Replica-moulded polydimethylsiloxane culture vessel lids attenuate osmotic drift in long-term cell cultures

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