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PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
We also successfully visualized the inhibitory effect of AxN on Aβ aggregation and deposition on SH-SY5Y human neuroblastoma cells using QDAβ nanoprobes (Figure 2).
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
QDAβ was specifically used to visualize Aβ aggregation with high sensitivity.
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
It also emits intense photostable signals, which allow the clear detection of Aβ aggregation over time, providing a precise and reliable method to monitor Aβ aggregation dynamics in live cells.
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
Similarly, Aβ fibril formation on SH-SY5Y cells was monitored using the ThT assay (Figure 3).
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
ThT is a specific dye that binds to the β-sheet of amyloid fibrils, increasing fluorescence intensity upon binding and assisting in the identification of fibril deposition .
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
Recent studies showed that AxN with 50 µM was able to inhibit 80% of Aβ fibril formation in rat pheochromocytoma (PC 12) cells and reduced ThT fluorescence, indicating the inhibitory ability of Aβ deposition .
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
These findings suggest that AxN inhibits Aβ aggregation and deposition by impeding fibril formation.
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
This confirms that AxN can attenuate the formation of Aβ fibrilization on neurons, which is a key feature of AD.
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
Several studies demonstrated the neuroprotective potential of AxN by mitigating Aβ-induced cytotoxicity in several cell lines.
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
The pre-treatment with 50 µM of AxN reduced cytotoxicity induced by 1 µM Aβ .
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
Similarly, another study demonstrated that 100 µM AxN had an anti-cytotoxic effect on olfactory ensheathing cells (OECs) against 10 µM Aβ-induced cytotoxicity .
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
Other studies also emphasized the ability of AxN to attenuate critical cellular pathways and inhibit programmed cell death .
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
In the current study, AxN’s protective effect was assessed using apoptosis markers, which showed a strong protective effect against early apoptosis but not against late necrosis (Figure 4).
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
Therefore, the results suggest that the neuroprotective effect of AxN may partially rescue programmed cell death rather than overall protection against cell death.
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
The accumulation and fibrillation of Aβ promoted cell death and neuronal damage in SH-SY5Y cells.
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
Neurotoxicity induced by Aβ aggregation influenced oxidative stress and mitochondrial dysfunction, leading to cell death and neuronal damage, which was inhibited by AxN in SH-SY5Y cells by regulating the apoptotic signaling pathway .
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
Previously, it was also shown that AxN notably reduced the percentage of apoptosis, but it could not reduce necrosis in PC12 and rat retinal ganglion cells (RGCs) .
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
This finding suggests that AxN may play a crucial role in mitigating neuronal damage and enhancing cell viability.
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
The ability to suppress cell apoptosis is considered a way to prevent AD.
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
A study reported that AxN is safe for human consumption without causing any harmful effects even at a high supplementation dose .
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
Considering these studies, it can be stated that AxN is a safe material that has a significant neuroprotective effect against cytotoxicity induced by Aβ fibril formation in several cell lines.
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
We also revealed that the cell motility inhibited by Aβ aggregation was reduced by AxN treatment (Figure 5 and Figure 6).
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
The WHA is a collective assay that involves cell–cell interactions and cell migration in a two-dimensional confluent cell monolayer .
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
WHA was previously performed on a monolayer of human brain microvascular endothelial cells (hBMECs) to evaluate the inductive effect of Aβ deposition, showing that Aβ deposition inhibited cell speed and distance covered during migration .
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
Previous studies showed that AxN enhanced migration in human keratinocyte (HaCaTs) cells.
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
It was also reported that AxN enhanced migration in the human keratinocyte cell line HaCaT, restored motility, improved effective cellular organization , and reduced migration in breast cancer cells, indicating its potential to limit metastasis .
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
These results suggest that AxN is not only an anti-inflammatory agent but also attenuates the reduced cell motility caused by Aβ aggregation in AD while acting as an anti-cancer compound.
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
Some limitations of this study need to be acknowledged.
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
First, as we mentioned, SH-SY5Y human neuroblastoma cells were treated with 25 µM for 24 h as an AD cell model.
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
This is the same Aβ concentration and aggregation time used previously to screen various inhibitors using the MSHTS system and has the advantage of shortening screening times by promoting aggregation with a high Aβ concentration.
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
However, the Aβ concentration of 25 µM is higher than physiological conditions, so this may not be a suitable AD cell model.
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
A future challenge will be to explore experimental conditions that more closely mimic physiological conditions.
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
In addition, although this study indicates an inhibition of the effect of AxN on necrosis and cell migration dynamics, there is a gap in understanding the exact mechanism, as findings are limited.
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
Further comprehensive studies are required to address the role and mechanism of the neuroprotective effects of AxN. Even though AxN showed promising results in cell models, it is challenging to study in vivo efficacy and its clinical application, as AxN has poor aqueous solubility and low oral bioavailability.
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
To overcome these barriers, formulation strategies like nano-capsulation and lipid-based cyclodextrin complexes can be considered to improve solubility and absorption.
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
However, it also requires limiting the formulation stability and safety concerns in the biological environment to ensure AxN’s clinical potential.
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
Human neuroblastoma SH-SY5Y cells were obtained from KAC (Kyoto, Japan).
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
Dulbecco’s Modified Eagle’s Medium (DMEM), penicillin-streptomycin, dimethyl sulfoxide (DMSO) and 1,1,1,3,3,3-hexafluoro-2-propanal (HFIP) (Fujifilm, Wako, Osaka, Japan), AxN (010-2761, Fujifilm, Wako, Osaka, Japan), DMEM/F12, fetal bovine serum (FBS) and fibronectin (Gibco/Life Technologies, New York, NY, USA), human Aβ42 (4349-v; Peptide Institute Inc., Osaka, Japan), Hoechst 33342 (H1399, Invitrogen, Carlsbad, CA, USA), pSIVA-IABND (N,N′-dimethyl-N-(iodoacetyl)-N′-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)ethylenediamine), and propidium iodide (PI) (APO004, Bio-Rad Laboratories, Inc., Tokyo, Japan) were purchased.
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
Quantum dot—Aβ40 (QDAβ) was prepared according to our previous reports .
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
SH-SY5Y cells were cultured in DMEM supplemented with 10% FBS and 1% penicillin-streptomycin.
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
The confluent culture was washed with 1xPBS, the adherent cells were detached using trypsin at 37 °C, and centrifuged at 2000 rpm for 5 min at 37 °C.
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
The required amount of cell solution was then dispensed to culture dishes and made up with DMEM solution.
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
5 mg of Aβ42 was dissolved using 5 mL of HFIP in a vial.
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
The vial was covered and left to stand for 1 h at room temperature, gently stirring every 10 min.
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
The solution was sonicated at 25 °C for 10 min to monomerize the peptide.
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
HFIP was allowed to evaporate on a clean bench.
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
After evaporation, the peptide was dissolved in DMSO.
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
The prepared 1 mM or 2.5 mM Aβ42 was dispensed into microtubes and stored at −80 °C.
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
The samples were used immediately after thawing and were not subjected to refreezing.
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
The MSHTS system was established according to our previous report .
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
Aβ was co-incubated with five different concentrations (final 0.32, 1.6, 8.0, 40, and 200 µM) of AxN in 2% DMSO and stained with QDAβ in two different MSHTS solvents (PBS and DMEM/F12), then incubated at 37 °C for 24 h in a 1536-well plate.
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
The QDAβ-Aβ42 aggregates that formed in each well were observed by an inverted fluorescence microscope (TE2000, Nikon, Tokyo, Japan) equipped with a color CCD camera (DP72, Olympus, Tokyo, Japan) and an objective lens (Plan Fluor 4x/0.13 PhL DL, Nikon).
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
SD values of fluorescence intensities of 40,000 pixels (200 × 200 pixels) around the central region of each well were measured by Image J 1.53e software (NIH, Bethesda, MD, USA).
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
The EC50 graph was drawn using Prism 9 GraphPad LLC (San Diego, CA, USA) software.
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
A 96-well plate was coated with 1 mg/mL fibronectin and incubated overnight at 37 °C and 5% CO2.
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
Using a micropipette, 10 × 10 SH-SY5Y cells were then seeded in the 96-well plate and incubated overnight at 37 °C and 5% CO2.
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
25 nM QDAβ was dissolved in DMEM/F12 and mixed with 50 µM Aβ.
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
AxN-Aβ solution was prepared to achieve concentrations (final 0.032, 0.16, 0.8, 4.0, and 20 µM of AxN with 25 µM of Aβ).
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
The solutions were dispensed to respective wells containing cultured cells and incubated at 37 °C and 5% CO2 for 24 h. Aggregation was observed using a Ti-E inverted fluorescence microscope equipped with a color CMOS camera (DS-Ri2; Nikon) and an objective lens (20x, Nikon).
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
The Mean Gray value of images was measured using ImageJ 1.53e software (NIH).
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
SH-SY5Y cells were seeded and incubated in a 1 mg/mL fibronectin-coated 96-well plate for 24 h at 37 °C and 5% CO2.
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
The confluent cell layer was treated with 25 µM Aβ alone and with 20 µM ThT-0.032–20 µM AxN and incubated 24 h at 37 °C, 5% CO2.
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
The solutions were removed and fresh DMEM/F12 was dispensed.
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
Readings were taken using a fluorescence microplate reader (SH-9000, Yamato, Tokyo, Japan) at 490 nm.
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
Images were captured using a Ti-E inverted fluorescence microscope combined with NIS-Elements C 4.51 software (Nikon) and analyzed using ImageJ 1.53e software.
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
SH-SY5Y cells were seeded in the 1 mg/mL fibronectin-coated 96-well plate and incubated overnight at 37 °C and 5% CO2.
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
AxN dilutions (0.032–20 µM in DMEM/F12) were prepared and mixed with Aβ containing pSIVA-IANBD (100x) and PI (200x), then incubated for 24 h at 37 °C and 5% CO2.
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
Using a Ti-E inverted fluorescence microscope equipped with a DS-Ri2 and color CMOS camera, observations were made, and images were captured and analyzed using ImageJ 1.53e software.
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
A 96-well plate was coated with 1 mg/mL fibronectin, then incubated overnight at 37 °C and 5% CO2.
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
SH-SY5Y cells were seeded in the 96-well plate using DMEM solution and incubated overnight at 37 °C and 5% CO2 until confluence was reached.
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
The cell layer was scraped using a sharp tip to make a wound, and 0.032–20 µM AxN containing 25 µM Aβ was added.
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
Using a Ti-E inverted fluorescence microscope equipped with a DS-Ri2 color CMOS camera, four randomly selected scraped areas were captured every 15 min during a 24 h observation period and analyzed using ImageJ 1.53e software.
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
A 96-well plate was coated with 1 mg/mL fibronectin, then incubated overnight at 37 °C and 5% CO2.
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
SH-SY5Y cells were seeded in the wells and incubated at 37 °C and 5% CO2 for 24 h. SH-SY5Y cells were stained using 0.2% Hoechst and incubated for 1 h. 0.032–20 µM AxN dilutions with 25 µM Aβ were dispensed into respective wells.
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
Using a Ti-E inverted fluorescence microscope equipped with a DS-Ri2 color CMOS camera (Nikon, Tokyo, Japan), four randomly selected scraped areas were captured every 15 min during a 24 h observation period.
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
Images were analyzed using Fiji ImageJ 1.54f software (NIH).
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
The directional persistence was measured by migration pass/total length, and cell speed was determined by total length/migration time.
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
Graphs were plotted using Microsoft Excel software.
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
The statistical data were analyzed using Microsoft Excel (version 2401).
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
All values are expressed as the mean ± SD.
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
Two-tailed independent Student’s t-tests were used to compare the two groups.
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
A p-value of less than 0.05 was considered statistically significant.
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
Our study demonstrated that co-incubation of AxN with 25 µM Aβ was able to attenuate the effect of Aβ aggregation in SH-SY5Y cells, while also reducing the effect of Aβ-induced impairments in cell motility.
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
Although AxN did not restore late necrosis, early apoptosis was significantly inhibited, demonstrating its potent neuroprotective ability.
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
This study used human neuroblastoma SH-SY5Y cells as a cell model.
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
Our results suggest that AxN could be a promising neuroprotective therapeutic agent in treating neurodegenerative diseases, such as AD.
PMC12608137
Elucidation of the Neuroprotective Effects of Astaxanthin Against Amyloid β Toxicity in the SH-SY5Y Human Neuroblastoma Cell Line
In the future, AxN may be one agent used to cure neurodegenerative diseases.
PMC12705714
An open-source screening platform accelerates discovery of drug combinations
Drug combinations are essential to modern medicine, but their discovery remains slow and inefficient as experimental complexity expands rapidly with each additional drug tested.
PMC12705714
An open-source screening platform accelerates discovery of drug combinations
Although modern liquid handling systems enable complex and highly customizable experimental designs, a lack of strategies integrating these technologies with combination-specific analytical methods has limited throughput.
PMC12705714
An open-source screening platform accelerates discovery of drug combinations
Here we introduce Combocat, an open-source and streamlined framework that combines acoustic liquid handling protocols with machine learning-based inference to achieve ultrahigh-throughput drug combination screening.
PMC12705714
An open-source screening platform accelerates discovery of drug combinations
Using Combocat, we generate a reference dataset of over 800 unique combinations in a dense 10 × 10 matrix format across multiple cell types, and use this to train a predictive model that accurately infers drug combination effects from sparse data, drastically reducing the number of experimental measurements required.
PMC12705714
An open-source screening platform accelerates discovery of drug combinations
As proof of concept, we screened 9,045 combinations in a neuroblastoma cell line—the largest number of combinations tested in a single cell line to date—achieved using minimal resources.
PMC12705714
An open-source screening platform accelerates discovery of drug combinations
By integrating advanced drug dispensing technologies with predictive computational modeling, Combocat provides a scalable solution to accelerate the discovery of novel drug combinations.
PMC12705714
An open-source screening platform accelerates discovery of drug combinations
The development of effective drug combinations is fundamental to modern therapeutic strategies, particularly for cancers and infectious diseases where single-agent therapies often fail to achieve sustained efficacy.
PMC12705714
An open-source screening platform accelerates discovery of drug combinations
Combining drugs can have a synergistic effect on drug efficacy, overcoming resistance mechanisms and reducing the toxicity of treatment by requiring lower individual doses of each drug.
PMC12705714
An open-source screening platform accelerates discovery of drug combinations
Synergy, in this context, is defined as a combination effect that exceeds expectations based on individual drug activities under established models like Bliss independence (Fig. 1a).Fig.
PMC12705714
An open-source screening platform accelerates discovery of drug combinations
1The Combocat platform for dense drug combination screens.a Practical representation of drug synergy, where the combined effect of two drugs exceeds the expected effect (“E”) based on models like additivity or Bliss independence.
PMC12705714
An open-source screening platform accelerates discovery of drug combinations
b Examples of traditional high-throughput screens employing small (e.g., 3 × 3), sparse, or asymmetric (e.g., 2 × 7) matrix formats, which limit dose density.
PMC12705714
An open-source screening platform accelerates discovery of drug combinations
c Combocat’s streamlined approach for dense and reproducible drug combination screening.
PMC12705714
An open-source screening platform accelerates discovery of drug combinations
The 384-well plate format contains Drugs 1 and 2 (represented by upper and lower triangles, respectively), with concentrations shown ranging from low (blue) to high (red).