Non-invasive monitoring of the osteogenic differentiation of human mesenchymal stem cells on a polycaprolactone scaffold using Raman imaging

Gao, Yu; Xu, Chen; Wang, Lianhui

doi:10.1039/c6ra11636a

Cited by 6 publications

(4 citation statements)

References 26 publications

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“…18 The PC1 large band around 1440 cm −1 is related to the CH stretching modes of lipids. 27 This results confirm the wellestablished knowledge that PEF affect the lipids. 28 The PC1 score for the 0 and the 500 V/cm group were, respectively, −5.28 ± 7.35 and −4.62 ± 5.3 (Figure 2B).…”

Section: ■ Results and Discussionsupporting

confidence: 89%

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Comprehensive Characterization of the Interaction between Pulsed Electric Fields and Live Cells by Confocal Raman Microspectroscopy

et al. 2017

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This study reports a comprehensive analysis of the effect of 100 μs electric pulses on the biochemical composition of live cells using a label-free approach, confocal Raman microspectroscopy. We investigated different regions of interest around the nucleus of the cells and the dose-effect relationship related to different electric pulse parameters. We also extended the study to another cell type. Membrane resealing was monitored by pulsing the cells in reversible or irreversible electropermeabilization condition at different temperatures. Our results confirmed a previous publication showing that proteins and lipids were highly impacted by the delivery of electric pulses. These chemical changes were similar in different locations around the cell nucleus. By sweeping the field magnitude, the number of electric pulses, or their repetition rate, the Raman signatures of live cells appeared to be related to the electropermeabilization state, verified by Yo-Pro-1 uptake. We also demonstrated that the chemical changes in the Raman signatures were cell-dependent even if common features were noticed between the two cell types used.

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Section: ■ Results and Discussionsupporting

confidence: 89%

“…These Raman bands are known to be associated with the phenylalanine amino acid, like the 1003 cm –1 Raman peak . The PC1 large band around 1440 cm –1 is related to the CH stretching modes of lipids . This results confirm the well-established knowledge that PEF affect the lipids .…”

Section: Resultssupporting

confidence: 85%

Comprehensive Characterization of the Interaction between Pulsed Electric Fields and Live Cells by Confocal Raman Microspectroscopy

et al. 2017

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“…1c shows how the major Raman peaks contributing to the LV1 appeared clearly at 1003 and 1658 cm −1 . The 1448 cm −1 stretching mode of CH vibration, mainly attributed to lipids 40 , was also part of the LV1 which is consistent with the known effect of the µsPEF on the lipid cell bilayer 11 . Statistical analysis of the LV1 scores ( Fig.…”

Section: Resultssupporting

confidence: 85%

Monitoring the molecular composition of live cells exposed to electric pulses via label-free optical methods

Azan

Grognot

García-Sánchez

et al. 2020

Sci Rep

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the permeabilization of the live cells membrane by the delivery of electric pulses has fundamental interest in medicine, in particular in tumors treatment by electrochemotherapy. Since underlying mechanisms are still not fully understood, we studied the impact of electric pulses on the biochemical composition of live cells thanks to label-free optical methods: confocal Raman microspectroscopy and terahertz microscopy. A dose effect was observed after cells exposure to different field intensities and a major impact on cell peptide/protein content was found. Raman measurements reveal that protein structure and/or environment are modified by the electric pulses while terahertz measurements suggest a leakage of proteins and other intracellular compounds. We show that Raman and terahertz modalities are a particularly attractive complement to fluorescence microscopy which is the reference optical technique in the case of electropermeabilization. finally, we propose an analytical model for the influx and efflux of non-permeant molecules through transiently (electro)permeabilized cell membranes. The main consequence of the delivery of high intensity pulsed electric fields (PEF) of very short duration on biological samples is the permeabilization of the plasma membrane 1. This interaction between PEF and biological samples, termed electropermeabilization or electroporation, has led to many applications in industry or medicine, in particular using 100 microsecond pulses (µsPEF). For instance, electrochemotherapy consists in the combination of tumor cells electropermeabilization and a chemotherapeutic agent such as bleomycin or cisplatin drugs 2,3. Electrochemotherapy is nowadays commonly used for the treatment of many cancer types: skin cancer 4 , breast cancer 5 , head and neck cancer 6 , pancreatic cancer 7 , etc. Although electropermeabilization has been studied for decades, the underlying mechanisms are still not fully understood. It is well established that the electric field induces an additive transmembrane potential to the resting transmembrane potential of cells 8. Molecular dynamics simulations 9,10 have demonstrated that the creation of pores into the membrane is a stochastic phenomenon: the increase of the external field amplitude increases the probability of formation of the pores. Therefore, to ensure that pores occur within the duration of the applied pulses, appropriated field amplitudes must be used, a situation that led to the definition of functional threshold amplitudes by the experimentalists. The destabilization of the cell membrane is also associated with long-term effects on the membrane, such as membrane disorder and a decrease of membrane elasticity, 11,12 that occur minutes after the PEF delivery. Recently, we considered biochemical modifications of the membrane as the major contributor to the electropermeabilization process. Mass spectrometry analysis of the chemical composition of a simple membrane model, Giant Unilamellar Vesicles (GUV), exposed to PEF showed the peroxidation of phospholipids ...

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“…PCL has been widely used in biodegradable scaffolds and drug containing implants for tissue engineering and cancer therapy. [22][23][24][25] As PCL is a hydrophobic polymer, dispersing hydrophilic Ag nanoparticles within the hydrophobic matrix is challenging. Fortunately, we have developed a method to fabricate inorganic nanoparticle (e.g.…”

Section: Synthesis Characterization and Antibacterial Effect Of Ag Omentioning

confidence: 99%

Fabrication of a silver octahedral nanoparticle-containing polycaprolactone nanocomposite for antibacterial bone scaffolds

et al. 2017

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Non-invasive monitoring of the osteogenic differentiation of human mesenchymal stem cells on a polycaprolactone scaffold using Raman imaging

Abstract: Raman imaging allows the non-invasive and label-free monitoring of the preferred osteogenic differentiation of human mesenchymal stem cells on the polycaprolactone scaffolds.

Cited by 6 publications

References 26 publications

Comprehensive Characterization of the Interaction between Pulsed Electric Fields and Live Cells by Confocal Raman Microspectroscopy

Comprehensive Characterization of the Interaction between Pulsed Electric Fields and Live Cells by Confocal Raman Microspectroscopy

Monitoring the molecular composition of live cells exposed to electric pulses via label-free optical methods

Fabrication of a silver octahedral nanoparticle-containing polycaprolactone nanocomposite for antibacterial bone scaffolds

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