Scientific Report: The Effect of Mannitol Concentration on Cell Viability
Abstract
The objective of this study was to investigate the effects of varying concentrations of mannitol (0%, 5%, and 10%) on cell viability. Mannitol, a sugar alcohol, is known for its osmotic properties and is commonly utilized in clinical settings to reduce intracranial pressure. This experiment evaluated the cytotoxic effects of mannitol on cultured cells through a standard viability assay. The outcomes indicated a concentration-dependent effect on cell viability, showing significant reductions in viability at higher concentrations.
Introduction
Mannitol is a six-carbon sugar alcohol that is widely used in medicine, especially in renal protection and as an osmotic diuretic. Its role in cell biology extends to influencing cell metabolism, proliferation, and viability through osmotic effects. This study aims to elucidate the cytotoxic effects of mannitol at three different concentrations: 0% (control), 5%, and 10%. We hypothesized that increasing concentrations of mannitol would correspond with decreased cell viability due to osmotic stress.
Materials and Methods
Materials
- Cell line: Human epithelial kidney cells (HEK293)
- Mannitol (purity ≥ 99%)
- Phosphate-buffered saline (PBS)
- Cell viability assay kit (MTT assay)
- Dimethyl sulfoxide (DMSO)
- Incubator (37°C, 5% CO2)
Experimental Design
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Preparation of Mannitol Solutions: Mannitol solutions were prepared to provide 0%, 5%, and 10% concentrations in PBS.
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Cell Culture: HEK293 cells were cultured in Dulbecco's Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and antibiotics.
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Treatment Groups: Cells were seeded into 96-well plates at a density of 1 × 10^4 cells/well. After 24 hours of attachment, cells were treated with 200 µL of mannitol solutions (0%, 5%, or 10%) for 24 hours.
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Viability Assessment: After incubation, cell viability was assessed using the MTT assay. Cells were incubated with MTT solution for 4 hours, followed by the addition of DMSO to dissolve formazan crystals. Absorbance was measured at 570 nm using a microplate reader.
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Statistical Analysis: Data were analyzed using ANOVA followed by Bonferroni post-hoc tests to determine statistical significance.
Results
Cell Viability
The mean absorbance values for each treatment group were as follows:
- 0% Mannitol: 1.2 ± 0.05 (control)
- 5% Mannitol: 0.85 ± 0.07 (p < 0.01 vs. 0%)
- 10% Mannitol: 0.45 ± 0.06 (p < 0.001 vs. 0% and 5%)
A significant decrease in cell viability was observed with increasing concentrations of mannitol. Control cells (0% mannitol) exhibited the highest viability, while cells treated with 10% mannitol demonstrated drastic reductions in viability.
Discussion
The results support the hypothesis that mannitol concentrations significantly impact cell viability. The 0% mannitol group served as the control and showed optimal cell survival. With 5% mannitol, there was a notable reduction in viability, indicating the onset of osmotic stress. The most severe effect was noted at 10% mannitol, where cell survival was drastically compromised.
These findings align with previous studies suggesting that high concentrations of mannitol lead to osmotic-induced cell shrinkage and potential apoptosis. Mannitol's mechanism likely involves changes in intracellular fluid balance, leading to cell injury or death.
Conclusion
This study demonstrates that increasing concentrations of mannitol are associated with reduced cell viability in HEK293 cells. While 0% and 5% concentrations had moderate effects, the 10% concentration resulted in significant cytotoxicity. Future studies may explore the underlying mechanisms of mannitol-induced cytotoxicity and its potential therapeutic applications in clinical settings.
References
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