Highly Sensitive Ratiometric Fluorescent Fiber Matrixes for Oxygen Sensing with Micrometer-Spatial Resolution

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Main Authors: Grasso, Giuliana, Onesto, Valentina, Forciniti, Stefania, Amone, Eliana D, Colella, Francesco, Pierantoni, Lara, Fama, Valeria, Gigli, Giuseppe, Reis, Rui L., Oliveira, Joaquim M., del Mercato, Loretta L.
Format: Preprint
Published: 2024
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author Grasso, Giuliana
Onesto, Valentina
Forciniti, Stefania
Amone, Eliana D
Colella, Francesco
Pierantoni, Lara
Fama, Valeria
Gigli, Giuseppe
Reis, Rui L.
Oliveira, Joaquim M.
del Mercato, Loretta L.
author_facet Grasso, Giuliana
Onesto, Valentina
Forciniti, Stefania
Amone, Eliana D
Colella, Francesco
Pierantoni, Lara
Fama, Valeria
Gigli, Giuseppe
Reis, Rui L.
Oliveira, Joaquim M.
del Mercato, Loretta L.
contents Oxygen (O2)-sensing matrices are promising tools for the live monitoring of extracellular O2 consumption levels in long-term cell cultures. In this study, ratiometric O2-sensing membranes were prepared by electrospinning, an easy, low-cost, scalable, and robust method for fabricating nanofibers. Poly(ε-caprolactone) and poly(dimethyl)siloxane polymers were blended with tris(4,7-diphenyl-1,10-phenanthroline) ruthenium(II) dichloride, which was used as the O2-sensing probe, and rhodamine B isothiocyanate, which was used as the reference dye. The functionalized scaffolds were morphologically characterized by scanning electron microscopy, and their physicochemical profiles were obtained by Fourier transform infrared spectroscopy, thermogravimetric analysis, and water contact angle measurement. The sensing capabilities were investigated by confocal laser scanning microscopy, performing photobleaching, reversibility, and calibration curve studies toward different dissolved O2 (DO) concentrations. Electrospun sensing nanofibers showed a high response to changes in DO concentrations in the physiological-pathological range from 0.5 to 20% and good stability under ratiometric imaging. In addition, the sensing systems were highly biocompatible for cell growth promoting adhesiveness and growth of three cancer cell lines, namely metastatic melanoma cell line SK-MEL2, breast cancer cell line MCF-7, and pancreatic ductal adenocarcinoma cell line Panc-1, thus recreating a suitable biological environment in vitro. These O2-sensing biomaterials can potentially measure alterations in cell metabolism caused by changes in ambient O2 content during drug testing/validation and tissue regeneration processes.
format Preprint
id arxiv_https___arxiv_org_abs_2405_06550
institution arXiv
publishDate 2024
record_format arxiv
spellingShingle Highly Sensitive Ratiometric Fluorescent Fiber Matrixes for Oxygen Sensing with Micrometer-Spatial Resolution
Grasso, Giuliana
Onesto, Valentina
Forciniti, Stefania
Amone, Eliana D
Colella, Francesco
Pierantoni, Lara
Fama, Valeria
Gigli, Giuseppe
Reis, Rui L.
Oliveira, Joaquim M.
del Mercato, Loretta L.
Biological Physics
Oxygen (O2)-sensing matrices are promising tools for the live monitoring of extracellular O2 consumption levels in long-term cell cultures. In this study, ratiometric O2-sensing membranes were prepared by electrospinning, an easy, low-cost, scalable, and robust method for fabricating nanofibers. Poly(ε-caprolactone) and poly(dimethyl)siloxane polymers were blended with tris(4,7-diphenyl-1,10-phenanthroline) ruthenium(II) dichloride, which was used as the O2-sensing probe, and rhodamine B isothiocyanate, which was used as the reference dye. The functionalized scaffolds were morphologically characterized by scanning electron microscopy, and their physicochemical profiles were obtained by Fourier transform infrared spectroscopy, thermogravimetric analysis, and water contact angle measurement. The sensing capabilities were investigated by confocal laser scanning microscopy, performing photobleaching, reversibility, and calibration curve studies toward different dissolved O2 (DO) concentrations. Electrospun sensing nanofibers showed a high response to changes in DO concentrations in the physiological-pathological range from 0.5 to 20% and good stability under ratiometric imaging. In addition, the sensing systems were highly biocompatible for cell growth promoting adhesiveness and growth of three cancer cell lines, namely metastatic melanoma cell line SK-MEL2, breast cancer cell line MCF-7, and pancreatic ductal adenocarcinoma cell line Panc-1, thus recreating a suitable biological environment in vitro. These O2-sensing biomaterials can potentially measure alterations in cell metabolism caused by changes in ambient O2 content during drug testing/validation and tissue regeneration processes.
title Highly Sensitive Ratiometric Fluorescent Fiber Matrixes for Oxygen Sensing with Micrometer-Spatial Resolution
topic Biological Physics
url https://arxiv.org/abs/2405.06550