Luminescence imaging and toxicity assessment of graphene quantum dots using in vitro models
Graphene quantum dots (GQDs) have been of high interest due to their size and optical characteristics, which improves when functional groups are added to their borders and defects. In this work, the in vitro toxicity of aqueous dispersion of GQDs (w/wo amino-functionalization) was investigated in tw...
- Autores:
-
De Falco, Anna
SANTA-HELENA, EDUARDA
T. Toloza, Carlos A.
Almeida, Joseany
Larrude, Dunieskys G.
Pereira Meirelles, Fatima Ventura
Gioda, Carolina
Aucelio, Ricardo Q.
Gioda, Adriana
- Tipo de recurso:
- Article of investigation
- Fecha de publicación:
- 2022
- Institución:
- Corporación Universidad de la Costa
- Repositorio:
- REDICUC - Repositorio CUC
- Idioma:
- eng
- OAI Identifier:
- oai:repositorio.cuc.edu.co:11323/9925
- Acceso en línea:
- https://hdl.handle.net/11323/9925
https://repositorio.cuc.edu.co/
- Palabra clave:
- Graphene quantum dots
Cardiomyoblast
Cytotoxicity
Fluorescence imaging
Yeast
- Rights
- embargoedAccess
- License
- Atribución-NoComercial 4.0 Internacional (CC BY-NC 4.0)
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Luminescence imaging and toxicity assessment of graphene quantum dots using in vitro models |
| title |
Luminescence imaging and toxicity assessment of graphene quantum dots using in vitro models |
| spellingShingle |
Luminescence imaging and toxicity assessment of graphene quantum dots using in vitro models Graphene quantum dots Cardiomyoblast Cytotoxicity Fluorescence imaging Yeast |
| title_short |
Luminescence imaging and toxicity assessment of graphene quantum dots using in vitro models |
| title_full |
Luminescence imaging and toxicity assessment of graphene quantum dots using in vitro models |
| title_fullStr |
Luminescence imaging and toxicity assessment of graphene quantum dots using in vitro models |
| title_full_unstemmed |
Luminescence imaging and toxicity assessment of graphene quantum dots using in vitro models |
| title_sort |
Luminescence imaging and toxicity assessment of graphene quantum dots using in vitro models |
| dc.creator.fl_str_mv |
De Falco, Anna SANTA-HELENA, EDUARDA T. Toloza, Carlos A. Almeida, Joseany Larrude, Dunieskys G. Pereira Meirelles, Fatima Ventura Gioda, Carolina Aucelio, Ricardo Q. Gioda, Adriana |
| dc.contributor.author.none.fl_str_mv |
De Falco, Anna SANTA-HELENA, EDUARDA T. Toloza, Carlos A. Almeida, Joseany Larrude, Dunieskys G. Pereira Meirelles, Fatima Ventura Gioda, Carolina Aucelio, Ricardo Q. Gioda, Adriana |
| dc.subject.proposal.eng.fl_str_mv |
Graphene quantum dots Cardiomyoblast Cytotoxicity Fluorescence imaging Yeast |
| topic |
Graphene quantum dots Cardiomyoblast Cytotoxicity Fluorescence imaging Yeast |
| description |
Graphene quantum dots (GQDs) have been of high interest due to their size and optical characteristics, which improves when functional groups are added to their borders and defects. In this work, the in vitro toxicity of aqueous dispersion of GQDs (w/wo amino-functionalization) was investigated in two different cellular models (S. cerevisiae and H9c2 cell line). Results in yeast suggest that when at up to 25 % volume concentration, the effect of all tested GQDs was only inhibitory, and, in both cellular models, the toxic effect is rigorously dose-dependent. The comparison of IC50 values of all the tested GQDs reveals no significant variations among them, pointing to non-carbonized citric acid as the more toxic precursor. The obtained data suggest that functionalization makes GQDs less toxic, being the one functionalized with thioacetamide slightly more toxic, followed by the ones functionalized with thiourea and glutathione, respectively. Results confirm that their toxicity is characteristics as a whole, and not as the sum of the toxicity of the precursors. In both models, concentrations up to 2 % showed no significant toxicity. Finally, fluorescence microscopy images suggest that GQDs interact with the cellular membrane and enter in the cell, manifesting fluorescent properties. |
| publishDate |
2022 |
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2022 |
| dc.date.accessioned.none.fl_str_mv |
2023-02-24T13:57:10Z |
| dc.date.available.none.fl_str_mv |
2023 2023-02-24T13:57:10Z |
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Artículo de revista |
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http://purl.org/coar/resource_type/c_2df8fbb1 |
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Text |
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info:eu-repo/semantics/article |
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http://purl.org/redcol/resource_type/ART |
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info:eu-repo/semantics/publishedVersion |
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http://purl.org/coar/version/c_970fb48d4fbd8a85 |
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Anna De Falco, Eduarda Santa-Helena, Carlos A. T. Toloza, Joseany M. S. Almeida, Dunieskys G. Larrude, Fatima Ventura Pereira Meirelles, Carolina Rosa Gioda, Ricardo Q. Aucelio & Adriana Gioda (2022) Luminescence imaging and toxicity assessment of graphene quantum dots using in vitro models, Fullerenes, Nanotubes and Carbon Nanostructures, 30:6, 657-666, DOI: 10.1080/1536383X.2021.1995367 |
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1536-383X |
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https://hdl.handle.net/11323/9925 |
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10.1080/1536383X.2021.1995367 |
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1536-4046 |
| dc.identifier.instname.spa.fl_str_mv |
Corporación Universidad de la Costa |
| dc.identifier.reponame.spa.fl_str_mv |
REDICUC - Repositorio CUC |
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https://repositorio.cuc.edu.co/ |
| identifier_str_mv |
Anna De Falco, Eduarda Santa-Helena, Carlos A. T. Toloza, Joseany M. S. Almeida, Dunieskys G. Larrude, Fatima Ventura Pereira Meirelles, Carolina Rosa Gioda, Ricardo Q. Aucelio & Adriana Gioda (2022) Luminescence imaging and toxicity assessment of graphene quantum dots using in vitro models, Fullerenes, Nanotubes and Carbon Nanostructures, 30:6, 657-666, DOI: 10.1080/1536383X.2021.1995367 1536-383X 10.1080/1536383X.2021.1995367 1536-4046 Corporación Universidad de la Costa REDICUC - Repositorio CUC |
| url |
https://hdl.handle.net/11323/9925 https://repositorio.cuc.edu.co/ |
| dc.language.iso.spa.fl_str_mv |
eng |
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eng |
| dc.relation.ispartofjournal.spa.fl_str_mv |
Fullerenes Nanotubes and Carbon Nanostructures |
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[1] Novoselov, K. S.; Geim, A. K.; Morozov, S. V.; Jiang, D.; Zhang, Y.; Dubonos, S.; V; Grigorieva, I. V.; Firsov, A. A. Electric Field Effect in Atomically Thin Carbon Films. Science. 2004, 306, 666–669. DOI: 10.1126/science.1102896. [2] Geim, A. K.; Novoselov, K. S. The Rise of Graphene. Nat. Mater. 2007, 6, 183–191. DOI: 10.1038/nmat1849. [3] Ponomarenko, L.; Schedin, F.; Katsnelson, M.; Yang, R.; Hill, E.; Novoselov, K.; Geim, A. Chaotic Dirac Billiard in Graphene Quantum Dots. Science 2008, 320, 356–358. DOI: 10.1126/science.1154663. [4] Shen, J.; Zhu, Y.; Chen, C.; Yang, X.; Li, C. Facile Preparation and Upconversion Luminescence of Graphene Quantum Dots. Chem Commun (Camb). 2011, 47, 2580–2582. DOI: 10.1039/c0cc04812g. [5] Li, L.; Wu, G.; Yang, G.; Peng, J.; Zhao, J.; Zhu, J. Focusing on Luminescent Graphene Quantum Dots: Current Status and Future Perspectives. Nanoscale. 2013, 5, 4015–4039. DOI: 10.1039/c3nr33849e. [6] Samra, K. S.; Manpreet.; Singh, A. Facile Synthesis of Graphene Quantum Dots and Their Optical Characterization. Fulleren Nanotube Carbon Nanostruct. 2021, 29, 638–642. DOI: 10.1080/1536383X.2021.1878152. [7] Guo, Z.; Cai, B.; Cao, Q.; Su, Y.; Li, M.; Hu, J.; Yang, Z.; Zhang, Y. Facile Synthesis of Amine-Functionalized Graphene Quantum Dots with Highly PH-Sensitive Photoluminescence. Fulleren Nanotube Carbon Nanostruct. 2017, 25, 704–709. DOI: 10.1080/1536383X.2017.1381952. [8] Chen, N.; He, Y.; Su, Y.; Li, X.; Huang, Q.; Wang, H.; Zhang, X.; Tai, R.; Fan, C. The Cytotoxicity of Cadmium-Based Quantum Dots. Biomaterials. 2012, 33, 1238–1244. DOI: 10.1016/j.biomaterials.2011.10.070. [9] Migita, S.; Moquin, A.; Fujishiro, H.; Himeno, S.; Maysinger, D.; Winnik, F.; Taniguchi, A. Quantum Dots Induce Heat Shock-Related Cytotoxicity at Intracellular Environment. In Vitro Cell Dev. Biol. Anim. 2014, 50, 367–372. DOI: 10.1007/s11626-013-9693-2. [10] Wang, F.; Gu, Z.; Lei, W.; Wang, W.; Xia, X.; Hao, Q. Graphene Quantum Dots as a Fluorescent Sensing Platform for Highly Efficient Detection of Copper(II) Ions. Sens. Actuators B. 2014, 190, 516–522. DOI: 10.1016/j.snb.2013.09.009. [11] Li, J.; Liu, F.; Shao, Q.; Min, Y.; Costa, M.; Yeow, E.; Xing, B. Enzyme-Responsive Cell-Penetrating Peptide Conjugated Mesoporous Silica Quantum Dot Nanocarriers for Controlled Release of Nucleus-Targeted Drug Molecules and Real-Time Intracellular Fluorescence Imaging of Tumor Cells. Adv. Healthc. Mater. 2014, 3, 1230–1239. DOI: 10.1002/adhm.201300613. [12] Zhou, N.; Hao, Z.; Zhao, X.; Maharjan, S.; Zhu, S.; Song, Y.; Yang, B.; Lu, L. A Novel Fluorescent Retrograde Neural Tracer: Cholera Toxin B Conjugated Carbon Dots. Nanoscale. 2015, 7, 15635–15642. DOI: 10.1039/c5nr04361a. [13] Lu, H.; Li, W.; Dong, H.; Wei, M. Graphene Quantum Dots for Optical Bioimaging. Small (Weinheim an Der Bergstrasse, Germany) 2019, 15, 1902136. DOI: 10.1002/smll.201902136. [14] Luo, P.; Sahu, S.; Yang, S.; Sonkar, S.; Wang, J.; Wang, H.; LeCroy, G.; Cao, L.; Sun, Y. Carbon “Quantum” Dots for Optical bioimaging. J. Mater. Chem. B. 2013, 1, 2116–2127. DOI: 10.1039/c3tb00018d. [15] Pang, S. A PH Sensitive Fluorescent Carbon Dots for Urea and Urease Detection. Fulleren Nanotube Carbon Nanostruct. 2020, 28, 752–760. DOI: 10.1080/1536383X.2020.1759039. [16] Dong, Y.; Chen, C.; Zheng, X.; Gao, L.; Cui, Z.; Yang, H.; Guo, C.; Chi, Y.; Li, C. M. One-Step and High Yield Simultaneous Preparation of Single- and Multi-Layer Graphene Quantum Dots from CX-72 Carbon Black. J. Mater. Chem. 2012, 22, 8764–8766. DOI: 10.1039/c2jm30658a. [17] Fan, R. J.; Sun, Q.; Zhang, L.; Zhang, Y.; Lu, A. H. Photoluminescent Carbon Dots Directly Derived from Polyethylene Glycol and Their Application for Cellular Imaging. Carbon. 2014, 71, 87–93. DOI: 10.1016/j.carbon.2014.01.016. [18] Thakur, M.; Pandey, S.; Mewada, A.; Patil, V.; Khade, M.; Goshi, E.; Sharon, M. Antibiotic Conjugated Fluorescent Carbon Dots as a Theranostic Agent for Controlled Drug Release, Bioimaging, and Enhanced Antimicrobial Activity. J. Drug. Deliv. 2014, 2014, 282193–282199. DOI: 10.1155/2014/282193. [19] Wang, Y.; Anilkumar, P.; Cao, L.; Liu, J.; Luo, P.; Tackett, K.; Sahu, S.; Wang, P.; Wang, X.; Sun, Y. Carbon Dots of Different Composition and Surface Functionalization: Cytotoxicity Issues Relevant to Fluorescence Cell Imaging. Exp. Biol. Med. (Maywood). 2011, 236, 1231–1238. DOI: 10.1258/ebm.2011.011132. [20] Yang, S.; Wang, X.; Wang, H.; Lu, F.; Luo, P.; Cao, L.; Meziani, M.; Liu, J.; Liu, Y.; Chen, M.; et al. Carbon Dots as Nontoxic and High-Performance Fluorescence Imaging Agents. J. Phys. Chem. C Nanomater. Interfaces. 2009, 113, 18110–18114. DOI: 10.1021/jp9085969. [21] Ruan, S.; Qian, J.; Shen, S.; Zhu, J.; Jiang, X.; He, Q.; Gao, H. A Simple One-Step Method to Prepare Fluorescent Carbon Dots and Their Potential Application in Non-Invasive Glioma Imaging. Nanoscale. 2014, 6, 10040–10047. DOI: 10.1039/c4nr02657h. [22] Jin, X.; Sun, X.; Chen, G.; Ding, L.; Li, Y.; Liu, Z.; Wang, Z.; Pan, W.; Hu, C.; Wang, J. PH-Sensitive Carbon Dots for the Visualization of Regulation of Intracellular PH inside Living Pathogenic Fungal Cells. Carbon. 2015, 81, 388–395. DOI: 10. 1016/j.carbon.2014.09.071. [23] Bagheri, Z.; Ehtesabi, H.; Hallaji, Z.; Latifi, H.; Behroodi, E. Investigation the Cytotoxicity and Photo-Induced Toxicity of Carbon Dot on Yeast Cell. Ecotoxicol. Environ. Saf. 2018, 161, 245–250. DOI: 10.1016/j.ecoenv.2018.05.071. [24] Toloza, C. A. T.; Khan, S.; Silva, R. L. D.; Romani, E. C.; Larrude, D. G.; Louro, S. R. W.; Freire, F. L.; Aucelio, R. Q. Photoluminescence Suppression Effect Caused by Histamine on Amino-Functionalized Graphene Quantum Dots with the Mediation of Fe3þ, Cu2þ, Eu3þ: Application in the Analysis of Spoiled Tuna Fish. Microchem. J. 2017, 133C, 448–459. DOI: 10.1016/j.microc.2017.04.013. [25] Kasemets, K.; Suppi, S.; Kunnis-Beres, K.; Kahru, A. Toxicity of € CuO Nanoparticles to Yeast Saccharomyces Cerevisiae BY4741 Wild-Type and Its Nine Isogenic Single-Gene Deletion Mutants. Chem. Res. Toxicol. 2013, 26, 356–367. DOI: 10.1021/ tx300467d. [26] Trindade, G. S.; Capella, M. A. M.; Capella, L. S.; AffonsoMitidieri, O. R.; Rumjanek, V. M. Differences in Sensitivity to UVC, UVB and UVA Radiation of a Multidrug-Resistant Cell Line Overexpressing P-Glycoprotein. Photochem. Photobiol. 1999, 69, 694–699. DOI: 10.1111/j.1751-1097.1999.tb03348.x. [27] Fasbender, S.; Zimmermann, L.; Cadeddu, R.; Luysberg, M.; Moll, B.; Janiak, C.; Heinzel, T.; Haas, R. The Low Toxicity of Graphene Quantum Dots is Reflected by Marginal Gene Expression Changes of Primary Human Hematopoietic Stem Cells. Sci. Rep. 2019, 9, 1–13. DOI: 10.1038/s41598-019-48567-6. [28] Chang, Y.; Yang, S.; Liu, J.; Dong, E.; Wang, Y.; Cao, A.; Liu, Y.; Wang, H. In Vitro Toxicity Evaluation of Graphene Oxide on A549 Cells. Toxicol. Lett. 2011, 200, 201–210. DOI: 10.1016/j.toxlet.2010.11.016. [29] Stratford, M.; Nebe-von-Caron, G.; Steels, H.; Novodvorska, M.; Ueckert, J.; Archer, D. Weak-Acid Preservatives: PH and Proton Movements in the Yeast Saccharomyces Cerevisiae. Int. J. Food. Microbiol. 2013, 161, 164–171. DOI: 10.1016/j.ijfoodmicro.2012.12.013. [30] Nielsen, M.; Arneborg, N. The Effect of Citric Acid and PH on Growth and Metabolism of Anaerobic Saccharomyces Cerevisiae and Zygosaccharomyces Bailii Cultures. Food Microbiol. 2007, 24, 101–105. DOI: 10.1016/j.fm.2006.03.005. [31] Tabish, T.; Pranjol, M.; Karadag, I.; Horsell, D.; Whatmore, J.; Zhang, S. Influence of Luminescent Graphene Quantum Dots on Trypsin Activity. Int. J. Nanomedicine. 2018, 13, 1525–1538. DOI: 10.2147/IJN.S155021. [32] Lee, B.; Hasan, M.; Lichthardt, D.; Gonzalez-Rodriguez, R.; Naumov, A. Manganese-Nitrogen and Gadolinium-Nitrogen Co-Doped Graphene Quantum Dots as Bimodal Magnetic Resonance and Fluorescence Imaging Nanoprobes. Nanotechnol. 2021, 32, 095103. DOI: 10.1088/1361-6528/abc642. [33] Hadad, C.; Gonzalez-Domınguez, J. M.; Armelloni, S.; Mattinzoli, D.; Ikehata, M.; Istif, A.; Ostric, A.; Cellesi, F.; Alfieri, C. M.; Messa, P.; et al. Graphene Quantum Dots: From Efficient Preparation to Safe Renal Excretion. Nano Res. 2021, 14, 674–683. DOI: 10.1007/s12274-020-3096-y. [34] Toloza, C.; Almeida, J.; Khan, S.; Dos Santos, Y.; da Silva, A.; Romani, E.; Larrude, D.; Freire, F.; Aucelio, R. Gold Nanoparticles Coupled with Graphene Quantum Dots in Organized Medium to Quantify Aminoglycoside anti-Biotics in Yellow Fever Vaccine after Solid Phase Extraction Using a Selective Imprinted Polymer. J. Pharm. Biomed. Anal. 2018, 158, 480–493. DOI: 10.1016/j.jpba.2018.05.050. [35] Toloza, C. A. T.; Khan, S.; Silva, R. L. D.; Romani, E. C.; Freire, F. L.; Aucelio, R. Q. Different Approaches for Sensing Captopril Based on Functionalized Graphene Quantum Dots as Photoluminescent Probe. J. Lumin. 2016, 179, 83–92. DOI: 10. 1016/j.jlumin.2016.06.055. |
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Atribución-NoComercial 4.0 Internacional (CC BY-NC 4.0) |
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Atribución-NoComercial 4.0 Internacional (CC BY-NC 4.0)Copyright © 2023 Informa UK Limitedhttps://creativecommons.org/licenses/by-nc/4.0/info:eu-repo/semantics/embargoedAccesshttp://purl.org/coar/access_right/c_f1cfDe Falco, AnnaSANTA-HELENA, EDUARDAT. Toloza, Carlos A.Almeida, JoseanyLarrude, Dunieskys G.Pereira Meirelles, Fatima VenturaGioda, CarolinaAucelio, Ricardo Q.Gioda, Adriana2023-02-24T13:57:10Z20232023-02-24T13:57:10Z2022Anna De Falco, Eduarda Santa-Helena, Carlos A. T. Toloza, Joseany M. S. Almeida, Dunieskys G. Larrude, Fatima Ventura Pereira Meirelles, Carolina Rosa Gioda, Ricardo Q. Aucelio & Adriana Gioda (2022) Luminescence imaging and toxicity assessment of graphene quantum dots using in vitro models, Fullerenes, Nanotubes and Carbon Nanostructures, 30:6, 657-666, DOI: 10.1080/1536383X.2021.19953671536-383Xhttps://hdl.handle.net/11323/992510.1080/1536383X.2021.19953671536-4046Corporación Universidad de la CostaREDICUC - Repositorio CUChttps://repositorio.cuc.edu.co/Graphene quantum dots (GQDs) have been of high interest due to their size and optical characteristics, which improves when functional groups are added to their borders and defects. In this work, the in vitro toxicity of aqueous dispersion of GQDs (w/wo amino-functionalization) was investigated in two different cellular models (S. cerevisiae and H9c2 cell line). Results in yeast suggest that when at up to 25 % volume concentration, the effect of all tested GQDs was only inhibitory, and, in both cellular models, the toxic effect is rigorously dose-dependent. The comparison of IC50 values of all the tested GQDs reveals no significant variations among them, pointing to non-carbonized citric acid as the more toxic precursor. The obtained data suggest that functionalization makes GQDs less toxic, being the one functionalized with thioacetamide slightly more toxic, followed by the ones functionalized with thiourea and glutathione, respectively. Results confirm that their toxicity is characteristics as a whole, and not as the sum of the toxicity of the precursors. In both models, concentrations up to 2 % showed no significant toxicity. Finally, fluorescence microscopy images suggest that GQDs interact with the cellular membrane and enter in the cell, manifesting fluorescent properties.11 páginasapplication/pdfengTaylor and Francis Ltd.United Stateshttps://www.tandfonline.com/doi/full/10.1080/1536383X.2021.1995367Luminescence imaging and toxicity assessment of graphene quantum dots using in vitro modelsArtículo de revistahttp://purl.org/coar/resource_type/c_2df8fbb1Textinfo:eu-repo/semantics/articlehttp://purl.org/redcol/resource_type/ARTinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/version/c_970fb48d4fbd8a85Fullerenes Nanotubes and Carbon Nanostructures[1] Novoselov, K. S.; Geim, A. K.; Morozov, S. V.; Jiang, D.; Zhang, Y.; Dubonos, S.; V; Grigorieva, I. V.; Firsov, A. A. Electric Field Effect in Atomically Thin Carbon Films. Science. 2004, 306, 666–669. DOI: 10.1126/science.1102896.[2] Geim, A. K.; Novoselov, K. S. The Rise of Graphene. Nat. Mater. 2007, 6, 183–191. DOI: 10.1038/nmat1849.[3] Ponomarenko, L.; Schedin, F.; Katsnelson, M.; Yang, R.; Hill, E.; Novoselov, K.; Geim, A. Chaotic Dirac Billiard in Graphene Quantum Dots. Science 2008, 320, 356–358. DOI: 10.1126/science.1154663.[4] Shen, J.; Zhu, Y.; Chen, C.; Yang, X.; Li, C. Facile Preparation and Upconversion Luminescence of Graphene Quantum Dots. Chem Commun (Camb). 2011, 47, 2580–2582. DOI: 10.1039/c0cc04812g.[5] Li, L.; Wu, G.; Yang, G.; Peng, J.; Zhao, J.; Zhu, J. Focusing on Luminescent Graphene Quantum Dots: Current Status and Future Perspectives. Nanoscale. 2013, 5, 4015–4039. DOI: 10.1039/c3nr33849e.[6] Samra, K. S.; Manpreet.; Singh, A. Facile Synthesis of Graphene Quantum Dots and Their Optical Characterization. Fulleren Nanotube Carbon Nanostruct. 2021, 29, 638–642. DOI: 10.1080/1536383X.2021.1878152.[7] Guo, Z.; Cai, B.; Cao, Q.; Su, Y.; Li, M.; Hu, J.; Yang, Z.; Zhang, Y. 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DOI: 10. 1016/j.jlumin.2016.06.055.666657630Graphene quantum dotsCardiomyoblastCytotoxicityFluorescence imagingYeastPublicationORIGINALLuminescence imaging and toxicity assessment of graphene quantum dots using in vitro models.pdfLuminescence imaging and toxicity assessment of graphene quantum dots using in vitro models.pdfArtículoapplication/pdf2235889https://repositorio.cuc.edu.co/bitstreams/688f53e3-3cbd-4b9a-a352-808ce8e04b25/download70221ed15e818f0717a2db22a8fc0b93MD51LICENSElicense.txtlicense.txttext/plain; charset=utf-814828https://repositorio.cuc.edu.co/bitstreams/652a718b-2196-434e-b70b-532a3a88757c/download2f9959eaf5b71fae44bbf9ec84150c7aMD52TEXTLuminescence imaging and toxicity assessment of graphene quantum dots using in vitro models.pdf.txtLuminescence imaging and toxicity assessment of graphene quantum dots using in vitro models.pdf.txtExtracted texttext/plain47591https://repositorio.cuc.edu.co/bitstreams/94b7fbd1-129d-46f4-ac88-379f5afcbd7b/downloada0138a58a00ccd58bf1a14f769a49b1eMD53THUMBNAILLuminescence imaging and toxicity assessment of graphene quantum dots using in vitro models.pdf.jpgLuminescence imaging and toxicity assessment of graphene quantum dots using in vitro models.pdf.jpgGenerated Thumbnailimage/jpeg10535https://repositorio.cuc.edu.co/bitstreams/6dc70617-baa5-4206-9573-a1b4af908bbd/downloaddea4164de7c33a2d5006c7c085169f5aMD5411323/9925oai:repositorio.cuc.edu.co:11323/99252024-09-17 14:11:19.621https://creativecommons.org/licenses/by-nc/4.0/Copyright © 2023 Informa UK Limitedopen.accesshttps://repositorio.cuc.edu.coRepositorio de la Universidad de la Costa 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ada en las Obras Colectivas.

b.	Distribuir copias o fonogramas de las Obras, exhibirlas públicamente, ejecutarlas públicamente y/o ponerlas a disposición pública, incluyéndolas como incorporadas en Obras Colectivas, según corresponda.

c.	Distribuir copias de las Obras Derivadas que se generen, exhibirlas públicamente, ejecutarlas públicamente y/o ponerlas a disposición pública.
Los derechos mencionados anteriormente pueden ser ejercidos en todos los medios y formatos, actualmente conocidos o que se inventen en el futuro. Los derechos antes mencionados incluyen el derecho a realizar dichas modificaciones en la medida que sean técnicamente necesarias para ejercer los derechos en otro medio o formatos, pero de otra manera usted no está autorizado para realizar obras derivadas. Todos los derechos no otorgados expresamente por el Licenciante quedan por este medio reservados, incluyendo pero sin limitarse a aquellos que se mencionan en las secciones 4(d) y 4(e).

4. Restricciones.
La licencia otorgada en la anterior Sección 3 está expresamente sujeta y limitada por las siguientes restricciones:

a.	Usted puede distribuir, exhibir públicamente, ejecutar públicamente, o poner a disposición pública la Obra sólo bajo las condiciones de esta Licencia, y Usted debe incluir una copia de esta licencia o del Identificador Universal de Recursos de la misma con cada copia de la Obra que distribuya, exhiba públicamente, ejecute públicamente o ponga a disposición pública. No es posible ofrecer o imponer ninguna condición sobre la Obra que altere o limite las condiciones de esta Licencia o el ejercicio de los derechos de los destinatarios otorgados en este documento. No es posible sublicenciar la Obra. Usted debe mantener intactos todos los avisos que hagan referencia a esta Licencia y a la cláusula de limitación de garantías. Usted no puede distribuir, exhibir públicamente, ejecutar públicamente, o poner a disposición pública la Obra con alguna medida tecnológica que controle el acceso o la utilización de ella de una forma que sea inconsistente con las condiciones de esta Licencia. Lo anterior se aplica a la Obra incorporada a una Obra Colectiva, pero esto no exige que la Obra Colectiva aparte de la obra misma quede sujeta a las condiciones de esta Licencia. Si Usted crea una Obra Colectiva, previo aviso de cualquier Licenciante debe, en la medida de lo posible, eliminar de la Obra Colectiva cualquier referencia a dicho Licenciante o al Autor Original, según lo solicitado por el Licenciante y conforme lo exige la cláusula 4(c).

b.	Usted no puede ejercer ninguno de los derechos que le han sido otorgados en la Sección 3 precedente de modo que estén principalmente destinados o directamente dirigidos a conseguir un provecho comercial o una compensación monetaria privada. El intercambio de la Obra por otras obras protegidas por derechos de autor, ya sea a través de un sistema para compartir archivos digitales (digital file-sharing) o de cualquier otra manera no será considerado como estar destinado principalmente o dirigido directamente a conseguir un provecho comercial o una compensación monetaria privada, siempre que no se realice un pago mediante una compensación monetaria en relación con el intercambio de obras protegidas por el derecho de autor.

c.	Si usted distribuye, exhibe públicamente, ejecuta públicamente o ejecuta públicamente en forma digital la Obra o cualquier Obra Derivada u Obra Colectiva, Usted debe mantener intacta toda la información de derecho de autor de la Obra y proporcionar, de forma razonable según el medio o manera que Usted esté utilizando: (i) el nombre del Autor Original si está provisto (o seudónimo, si fuere aplicable), y/o (ii) el nombre de la parte o las partes que el Autor Original y/o el Licenciante hubieren designado para la atribución (v.g., un instituto patrocinador, editorial, publicación) en la información de los derechos de autor del Licenciante, términos de servicios o de otras formas razonables; el título de la Obra si está provisto; en la medida de lo razonablemente factible y, si está provisto, el Identificador Uniforme de Recursos (Uniform Resource Identifier) que el Licenciante especifica para ser asociado con la Obra, salvo que tal URI no se refiera a la nota sobre los derechos de autor o a la información sobre el licenciamiento de la Obra; y en el caso de una Obra Derivada, atribuir el crédito identificando el uso de la Obra en la Obra Derivada (v.g., "Traducción Francesa de la Obra del Autor Original," o "Guión Cinematográfico basado en la Obra original del Autor Original"). Tal crédito puede ser implementado de cualquier forma razonable; en el caso, sin embargo, de Obras Derivadas u Obras Colectivas, tal crédito aparecerá, como mínimo, donde aparece el crédito de cualquier otro autor comparable y de una manera, al menos, tan destacada como el crédito de otro autor comparable.

d.	Para evitar toda confusión, el Licenciante aclara que, cuando la obra es una composición musical:

i.	Regalías por interpretación y ejecución bajo licencias generales. El Licenciante se reserva el derecho exclusivo de autorizar la ejecución pública o la ejecución pública digital de la obra y de recolectar, sea individualmente o a través de una sociedad de gestión colectiva de derechos de autor y derechos conexos (por ejemplo, SAYCO), las regalías por la ejecución pública o por la ejecución pública digital de la obra (por ejemplo Webcast) licenciada bajo licencias generales, si la interpretación o ejecución de la obra está primordialmente orientada por o dirigida a la obtención de una ventaja comercial o una compensación monetaria privada.

ii.	Regalías por Fonogramas. El Licenciante se reserva el derecho exclusivo de recolectar, individualmente o a través de una sociedad de gestión colectiva de derechos de autor y derechos conexos (por ejemplo, los consagrados por la SAYCO), una agencia de derechos musicales o algún agente designado, las regalías por cualquier fonograma que Usted cree a partir de la obra (“versión cover”) y distribuya, en los términos del régimen de derechos de autor, si la creación o distribución de esa versión cover está primordialmente destinada o dirigida a obtener una ventaja comercial o una compensación monetaria privada.

e.	Gestión de Derechos de Autor sobre Interpretaciones y Ejecuciones Digitales (WebCasting). Para evitar toda confusión, el Licenciante aclara que, cuando la obra sea un fonograma, el Licenciante se reserva el derecho exclusivo de autorizar la ejecución pública digital de la obra (por ejemplo, webcast) y de recolectar, individualmente o a través de una sociedad de gestión colectiva de derechos de autor y derechos conexos (por ejemplo, ACINPRO), las regalías por la ejecución pública digital de la obra (por ejemplo, webcast), sujeta a las disposiciones aplicables del régimen de Derecho de Autor, si esta ejecución pública digital está primordialmente dirigida a obtener una ventaja comercial o una compensación monetaria privada.

5. Representaciones, Garantías y Limitaciones de Responsabilidad.
A MENOS QUE LAS PARTES LO ACORDARAN DE OTRA FORMA POR ESCRITO, EL LICENCIANTE OFRECE LA OBRA (EN EL ESTADO EN EL QUE SE ENCUENTRA) “TAL CUAL”, SIN BRINDAR GARANTÍAS DE CLASE ALGUNA RESPECTO DE LA OBRA, YA SEA EXPRESA, IMPLÍCITA, LEGAL O CUALQUIERA OTRA, INCLUYENDO, SIN LIMITARSE A ELLAS, GARANTÍAS DE TITULARIDAD, COMERCIABILIDAD, ADAPTABILIDAD O ADECUACIÓN A PROPÓSITO DETERMINADO, AUSENCIA DE INFRACCIÓN, DE AUSENCIA DE DEFECTOS LATENTES O DE OTRO TIPO, O LA PRESENCIA O AUSENCIA DE ERRORES, SEAN O NO DESCUBRIBLES (PUEDAN O NO SER ESTOS DESCUBIERTOS). ALGUNAS JURISDICCIONES NO PERMITEN LA EXCLUSIÓN DE GARANTÍAS IMPLÍCITAS, EN CUYO CASO ESTA EXCLUSIÓN PUEDE NO APLICARSE A USTED.

6. Limitación de responsabilidad.
A MENOS QUE LO EXIJA EXPRESAMENTE LA LEY APLICABLE, EL LICENCIANTE NO SERÁ RESPONSABLE ANTE USTED POR DAÑO ALGUNO, SEA POR RESPONSABILIDAD EXTRACONTRACTUAL, PRECONTRACTUAL O CONTRACTUAL, OBJETIVA O SUBJETIVA, SE TRATE DE DAÑOS MORALES O PATRIMONIALES, DIRECTOS O INDIRECTOS, PREVISTOS O IMPREVISTOS PRODUCIDOS POR EL USO DE ESTA LICENCIA O DE LA OBRA, AUN CUANDO EL LICENCIANTE HAYA SIDO ADVERTIDO DE LA POSIBILIDAD DE DICHOS DAÑOS. ALGUNAS LEYES NO PERMITEN LA EXCLUSIÓN DE CIERTA RESPONSABILIDAD, EN CUYO CASO ESTA EXCLUSIÓN PUEDE NO APLICARSE A USTED.

7. Término.

a.	Esta Licencia y los derechos otorgados en virtud de ella terminarán automáticamente si Usted infringe alguna condición establecida en ella. Sin embargo, los individuos o entidades que han recibido Obras Derivadas o Colectivas de Usted de conformidad con esta Licencia, no verán terminadas sus licencias, siempre que estos individuos o entidades sigan cumpliendo íntegramente las condiciones de estas licencias. Las Secciones 1, 2, 5, 6, 7, y 8 subsistirán a cualquier terminación de esta Licencia.

b.	Sujeta a las condiciones y términos anteriores, la licencia otorgada aquí es perpetua (durante el período de vigencia de los derechos de autor de la obra). No obstante lo anterior, el Licenciante se reserva el derecho a publicar y/o estrenar la Obra bajo condiciones de licencia diferentes o a dejar de distribuirla en los términos de esta Licencia en cualquier momento; en el entendido, sin embargo, que esa elección no servirá para revocar esta licencia o que deba ser otorgada , bajo los términos de esta licencia), y esta licencia continuará en pleno vigor y efecto a menos que sea terminada como se expresa atrás. La Licencia revocada continuará siendo plenamente vigente y efectiva si no se le da término en las condiciones indicadas anteriormente.

8. Varios.

a.	Cada vez que Usted distribuya o ponga a disposición pública la Obra o una Obra Colectiva, el Licenciante ofrecerá al destinatario una licencia en los mismos términos y condiciones que la licencia otorgada a Usted bajo esta Licencia.

b.	Si alguna disposición de esta Licencia resulta invalidada o no exigible, según la legislación vigente, esto no afectará ni la validez ni la aplicabilidad del resto de condiciones de esta Licencia y, sin acción adicional por parte de los sujetos de este acuerdo, aquélla se entenderá reformada lo mínimo necesario para hacer que dicha disposición sea válida y exigible.

c.	Ningún término o disposición de esta Licencia se estimará renunciada y ninguna violación de ella será consentida a menos que esa renuncia o consentimiento sea otorgado por escrito y firmado por la parte que renuncie o consienta.

d.	Esta Licencia refleja el acuerdo pleno entre las partes respecto a la Obra aquí licenciada. No hay arreglos, acuerdos o declaraciones respecto a la Obra que no estén especificados en este documento. El Licenciante no se verá limitado por ninguna disposición adicional que pueda surgir en alguna comunicación emanada de Usted. Esta Licencia no puede ser modificada sin el consentimiento mutuo por escrito del Licenciante y Usted.
 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