Spheroidal Model of SKBR3 and U87MG Cancer Cells for Live Imaging of Caspase-3 during Apoptosis Induced by Singlet Oxygen in Photodynamic Therapy

Pevná, Viktória; Máčajová, Mariana; Hovan, Andrej; Bánó, Gregor; Meta, Majlinda; Bilčı́k, Boris; Palková, Júlia; Huntošová, Veronika

doi:10.3390/biomedicines10092141

Cited by 6 publications

(5 citation statements)

References 45 publications

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“…To further evaluate the capability of as-synthesized pMAD hydrogel for MRI, pMAD5 with the highest APT-weighted CEST effect in Z-spectrum (Figure b) was selected to prepare a tumor spheroid phantom. Briefly, U87 spheroids were prepared based on the literature . U87 spheroids (Figure S9) were placed in the glass vial for MRI measurement.…”

Section: Resultsmentioning

confidence: 99%

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Formation of Zwitterionic and Self-Healable Hydrogels via Amino-yne Click Chemistry for Development of Cellular Scaffold and Tumor Spheroid Phantom for MRI

Nguyen,

Chow,

Nguyen

et al. 2024

ACS Appl. Mater. Interfaces

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In situ-forming biocompatible hydrogels have great potential in various medical applications. Here, we introduce a pHresponsive, self-healable, and biocompatible hydrogel for cell scaffolds and the development of a tumor spheroid phantom for magnetic resonance imaging. The hydrogel (pMAD) was synthesized via amino-yne click chemistry between poly(2-methacryloyloxyethyl phosphorylcholine-co-2-aminoethylmethacrylamide) and dialkyne polyethylene glycol. Rheology analysis, compressive mechanical testing, and gravimetric analysis were employed to investigate the gelation time, mechanical properties, equilibrium swelling, and degradability of pMAD hydrogels. The reversible enamine and imine bond mechanisms leading to the sol-to-gel transition in acidic conditions (pH ≤ 5) were observed. The pMAD hydrogel demonstrated potential as a cellular scaffold, exhibiting high viability and NIH-3T3 fibroblast cell encapsulation under mild conditions (37 °C, pH 7.4). Additionally, the pMAD hydrogel also demonstrated the capability for in vitro magnetic resonance imaging of glioblastoma tumor spheroids based on the chemical exchange saturation transfer effect. Given its advantages, the pMAD hydrogel emerges as a promising material for diverse biomedical applications, including cell carriers, bioimaging, and therapeutic agent delivery.

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Section: Resultsmentioning

confidence: 99%

“…Briefly, U87 spheroids were prepared based on the literature. 59 U87 spheroids ( Figure S9 ) were placed in the glass vial for MRI measurement. This heightened sensitivity to proton transfer enables pMAD5 for the CEST MRI experiment, emphasizing its potential for enhanced molecular imaging.…”

Section: Resultsmentioning

confidence: 99%

Formation of Zwitterionic and Self-Healable Hydrogels via Amino-yne Click Chemistry for Development of Cellular Scaffold and Tumor Spheroid Phantom for MRI

Nguyen,

Chow,

Nguyen

et al. 2024

ACS Appl. Mater. Interfaces

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“…These regimens were chosen to modulate the metabolic activity of mitochondria stressed with the known protein kinase C inhibitor hypericin in the dark and the activator after light application [ 21 , 33 , 34 ]. Hypericin has been used in PDT as a photosensitizer with promising properties for diagnosing and curing cancer with PDT [ 45 , 46 , 47 , 48 , 49 , 50 , 51 ]. The main advantages of PDT are its high cancer specificity and minimal toxicity compared with healthy cells.…”

Section: Resultsmentioning

confidence: 99%

The Remodulation of Actin Bundles during the Stimulation of Mitochondria in Adult Human Fibroblasts in Response to Light

Olejárová,

Horváth,

Huntošová

2023

Pharmaceutics

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β-actin belongs to cytoskeletal structures that change dynamically in cells according to various stimuli. Human skin can be considered as an organ that is very frequently exposed to various stress factors, of which light plays an important role. The present study focuses on adult human fibroblasts exposed to two types of light stress. Orange light with a wavelength of 590 nm was used here to stimulate the photosensitizer localized in the cells as a residual dose of photodynamic therapy (PDT). On the other hand, near-infrared light with a wavelength of 808 nm was considered for photobiomodulation (PBM), which is often used in healing processes. Confocal fluorescence microscopy was used to observe changes in intercellular communication, mitochondrial structures, and cytoskeletal dynamics defined by the remodulation of β-actin of fibroblasts. The number of β-actin bundles forming spherical structures was detected after light exposure. These structures as β-actin oligomers were confirmed with super-resolution microscopy. While PDT led to the disintegration of actin oligomers, PBM increased their number. The interaction of β-actin with mitochondria was observed. The combination of PDT and PBM treatments is important to minimize the side effects of cancer treatment with PDT on healthy cells, as shown by the cell metabolism assay in this work. In this work, β-actin is presented as an important parameter that changes and is involved in the response of cells to PDT and PBM.

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“…We investigate singlet oxygen generation in U87MG glioblastoma cells using hypericin as a photosensitizer, which we have extensively studied over the last two decades. [22][23][24][25] Hypericin is not water soluble and aggregates in a water environment. 26 Aggregated hypericin molecules can no longer generate singlet oxygen.…”

Section: Introductionmentioning

confidence: 99%

“…The present work extends the singlet oxygen lifetime studies to another cell/photosensitizer system. We investigate singlet oxygen generation in U87MG glioblastoma cells using hypericin as a photosensitizer, which we have extensively studied over the last two decades 22–25 . Hypericin is not water soluble and aggregates in a water environment 26 .…”

Section: Introductionmentioning

confidence: 99%

Singlet oxygen lifetime changes in dying glioblastoma cells

Hovan

Pevná

Huntošová

et al. 2023

Photochem & Photobiology

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Time‐resolved phosphorescence detection was employed to determine the lifetime of singlet oxygen in live cells. Using hypericin as a photosensitizer, singlet oxygen was generated in U87MG glioblastoma cells. The phosphorescence of singlet oxygen was detected in aqueous cell suspensions following pulsed laser excitation. Our goal was to eliminate or reduce the problems associated with lifetime measurements in water‐based cell suspensions. The apparatus enabled simultaneous singlet oxygen phosphorescence and transient absorption measurements, reducing uncertainty in lifetime estimation. The changes in singlet oxygen lifetime were observed during early and late apoptosis induced by photodynamic action. Our findings show that the effective lifetime of singlet oxygen in the intracellular space of the studied glioblastoma cells is 0.4 μs and increases to 1.5 μs as apoptosis progresses. Another group of hypericin, presumably located in the membrane blebs and the plasma membrane of apoptotic cells, generates singlet oxygen with a lifetime of 1.9 μs.

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Spheroidal Model of SKBR3 and U87MG Cancer Cells for Live Imaging of Caspase-3 during Apoptosis Induced by Singlet Oxygen in Photodynamic Therapy

Cited by 6 publications

References 45 publications

Formation of Zwitterionic and Self-Healable Hydrogels via Amino-yne Click Chemistry for Development of Cellular Scaffold and Tumor Spheroid Phantom for MRI

Formation of Zwitterionic and Self-Healable Hydrogels via Amino-yne Click Chemistry for Development of Cellular Scaffold and Tumor Spheroid Phantom for MRI

The Remodulation of Actin Bundles during the Stimulation of Mitochondria in Adult Human Fibroblasts in Response to Light

Singlet oxygen lifetime changes in dying glioblastoma cells

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