Quantifying Cytosolic Cytochrome c Concentration Using Carbon Quantum Dots as a Powerful Method for Apoptosis Detection.
apoptosis
carbon dots
cells
cytochrome c
fluorescence
Journal
Pharmaceutics
ISSN: 1999-4923
Titre abrégé: Pharmaceutics
Pays: Switzerland
ID NLM: 101534003
Informations de publication
Date de publication:
25 Sep 2021
25 Sep 2021
Historique:
received:
05
08
2021
revised:
08
09
2021
accepted:
20
09
2021
entrez:
23
10
2021
pubmed:
24
10
2021
medline:
24
10
2021
Statut:
epublish
Résumé
Cytochrome c (Cyt c) is a key biomarker for early apoptosis, and many methods were designed to detect its release from mitochondria. For a proper evaluation of these programed cell death mechanisms, fluorescent nanoparticles are excellent candidates due to their valuable optical properties. Among all classes of nanoparticles developed thus far, carbon-based quantum dots bring qualitative and efficient imaging strategies for biomedical applications as a consequence of their biocompatibility and low cytotoxicity. In this study, we synthesized carbon quantum dots smaller than 5 nm from sodium citrate and polyethylene imine. These nanoparticles were rigorously characterized, and their quenching capacity in apoptotic events was assessed in A549 cells treated with staurosporine and etoposide. For the evaluation of Cyt c release, a phenomenon directly correlated with apoptotic events, we ran a semiquantitative analysis using confocal laser scanning microscopy. Carbon quantum dots were synthesized and were successfully employed for Cyt c detection by means of fluorescence microscopy. Significant drops in fluorescence intensity were observed in the case of cells treated with apoptosis-inducing therapeutic compounds compared to untreated cells, confirming Cyt c release from mitochondria to cytosol. Considering these results, we strongly believe this method can contribute to an indirect in vitro evaluation of apoptosis.
Sections du résumé
BACKGROUND
BACKGROUND
Cytochrome c (Cyt c) is a key biomarker for early apoptosis, and many methods were designed to detect its release from mitochondria. For a proper evaluation of these programed cell death mechanisms, fluorescent nanoparticles are excellent candidates due to their valuable optical properties. Among all classes of nanoparticles developed thus far, carbon-based quantum dots bring qualitative and efficient imaging strategies for biomedical applications as a consequence of their biocompatibility and low cytotoxicity.
METHODS
METHODS
In this study, we synthesized carbon quantum dots smaller than 5 nm from sodium citrate and polyethylene imine. These nanoparticles were rigorously characterized, and their quenching capacity in apoptotic events was assessed in A549 cells treated with staurosporine and etoposide. For the evaluation of Cyt c release, a phenomenon directly correlated with apoptotic events, we ran a semiquantitative analysis using confocal laser scanning microscopy.
RESULTS
RESULTS
Carbon quantum dots were synthesized and were successfully employed for Cyt c detection by means of fluorescence microscopy. Significant drops in fluorescence intensity were observed in the case of cells treated with apoptosis-inducing therapeutic compounds compared to untreated cells, confirming Cyt c release from mitochondria to cytosol.
CONCLUSION
CONCLUSIONS
Considering these results, we strongly believe this method can contribute to an indirect in vitro evaluation of apoptosis.
Identifiants
pubmed: 34683849
pii: pharmaceutics13101556
doi: 10.3390/pharmaceutics13101556
pmc: PMC8537359
pii:
doi:
Types de publication
Journal Article
Langues
eng
Subventions
Organisme : Unitatea Executiva pentru Finantarea Invatamantului Superior, a Cercetarii, Dezvoltarii si Inovarii
ID : PN-III-P4-ID-PCCF-2016-0112
Organisme : "Iuliu Hațieganu" University of Medicine and Pharmacy, Cluj-Napoca, Romania
ID : 1529/46 PCD
Références
Wiley Interdiscip Rev Nanomed Nanobiotechnol. 2020 Jul;12(4):e1617
pubmed: 32040882
Biosens Bioelectron. 2017 Dec 15;98:415-420
pubmed: 28711028
ACS Nano. 2017 Mar 28;11(3):2313-2381
pubmed: 28290206
Cell Death Differ. 2003 Jul;10(7):853-5
pubmed: 12815469
Sci Rep. 2019 Sep 30;9(1):14004
pubmed: 31570739
Nanoscale Res Lett. 2019 Aug 13;14(1):272
pubmed: 31410663
Nanomaterials (Basel). 2019 Nov 15;9(11):
pubmed: 31731719
Toxins (Basel). 2018 May 11;10(5):
pubmed: 29751687
Int J Biol Macromol. 2019 Jul 1;132:316-329
pubmed: 30940582
Oxid Med Cell Longev. 2014;2014:678371
pubmed: 25215174
Sci Rep. 2016 Sep 26;6:34064
pubmed: 27666530
Nanomaterials (Basel). 2019 Oct 18;9(10):
pubmed: 31635415
Physiol Rev. 2007 Jan;87(1):99-163
pubmed: 17237344
J Colloid Interface Sci. 2015 Jan 1;437:17-23
pubmed: 25310578
Nanoscale. 2018 Mar 15;10(11):5342-5349
pubmed: 29509193
Crit Rev Anal Chem. 2016 Nov;46(6):502-20
pubmed: 26941009
Sci Rep. 2018 Oct 22;8(1):15593
pubmed: 30348974
Sci Rep. 2015 Dec 22;5:18807
pubmed: 26689549
Photochem Photobiol. 1969 May;9(5):439-44
pubmed: 5771430
BMC Biol. 2014 May 27;12:34
pubmed: 24884669
EXCLI J. 2015 Jan 19;14:95-108
pubmed: 26600742
Langmuir. 2019 Jul 16;35(28):9115-9132
pubmed: 31267753
Oncogene. 2001 Jun 7;20(26):3354-62
pubmed: 11423986
ACS Appl Mater Interfaces. 2018 Aug 1;10(30):25166-25173
pubmed: 29979027
Exp Cell Res. 2002 Nov 1;280(2):233-43
pubmed: 12413889
Chem Soc Rev. 2018 Sep 17;47(18):6930-6946
pubmed: 30062349
Methods Mol Biol. 2019;2000:227-234
pubmed: 31148018
Cytometry A. 2006 Jun;69(6):515-23
pubmed: 16680678
Chem Rev. 2015 Oct 14;115(19):10816-906
pubmed: 25997028
Cell Death Differ. 2006 Sep;13(9):1423-33
pubmed: 16676004
BMC Cancer. 2009 Jun 08;9:174
pubmed: 19500428
Int J Mol Sci. 2018 Aug 25;19(9):
pubmed: 30149619
Nat Rev Mol Cell Biol. 2010 Sep;11(9):621-32
pubmed: 20683470