In vitro cytotoxicity of the ternary PAMAM G3–pyridoxal–biotin bioconjugate

In vitro cytotoxicity of the ternary PAMAM G3–pyridoxal–biotin bioconjugate

Łukasz Uram, Magdalena Szuster, Krzysztof Gargasz, Aleksandra Filipowicz, Elżbieta Wałajtys-Rode, Stanisław Wołowiec Cosmetology Department, University of Information Technology and Management in Rzeszów, Rzeszów, Poland Abstract: A third-generation polyamidoamine dendrimer (...

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Autores principales: Uram Ł, Szuster M, Gargasz K, Filipowicz A, Wałajtys-Rode E, Wołowiec S
Formato: article
Lenguaje:EN
Publicado: Dove Medical Press 2013
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Medicine (General)
R5-920
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spelling oai:doaj.org-article:0eaea7dcd7e049578554a9b1654d78192021-12-02T07:28:37ZIn vitro cytotoxicity of the ternary PAMAM G3–pyridoxal–biotin bioconjugate1178-2013https://doaj.org/article/0eaea7dcd7e049578554a9b1654d78192013-12-01T00:00:00Zhttp://www.dovepress.com/in-vitro-cytotoxicity-of-the-ternary-pamam-g3ndashpyridoxalndashbiotin-a15264https://doaj.org/toc/1178-2013 Łukasz Uram, Magdalena Szuster, Krzysztof Gargasz, Aleksandra Filipowicz, Elżbieta Wałajtys-Rode, Stanisław Wołowiec Cosmetology Department, University of Information Technology and Management in Rzeszów, Rzeszów, Poland Abstract: A third-generation polyamidoamine dendrimer (PAMAM G3) was used as a macromolecular carrier for pyridoxal and biotin. The binary covalent bioconjugate of G3, with nine molecules of biotin per one molecule of G3 (G39B), and the ternary covalent bioconjugate of G3, with nine biotin and ten pyridoxal molecules (G39B10P), were synthesized. The biotin and pyridoxal residues of the bioconjugate were available for carboxylase and transaminase enzymes, as demonstrated in the conversion of pyruvate to oxaloacetate and alanine to pyruvate, respectively, by in vitro monitoring of the reactions, using 1H nuclear magnetic resonance spectroscopy. The toxicity of the ternary bioconjugate (BC-PAMAM) was studied in vitro on BJ human normal skin fibroblasts and human squamous cell carcinoma (SCC-15) cell cultures in comparison with PAMAM G3, using three cytotoxicity assays (XTT, neutral red, and crystal violet) and an estimation of apoptosis by confocal microscopy detection. The tests have shown that BC-PAMAM has significantly lower cytotoxicity compared with PAMAM. Nonconjugated PAMAM was not cytotoxic at concentrations up to 5 µM (NR) and 10 µM (XTT), and BC-PAMAM was not cytotoxic up to 50 µM (both assays) for both cell lines. It has been also found that normal fibroblasts were more sensitive than SCC to both PAMAM and BC-PAMAM. The effect of PAMAM and BC-PAMAM on the initiation of apoptosis (PAMAM in fibroblasts at 5 µM and BC-PAMAM at 10 µM in both cell lines) corresponded with cytotoxicity assays for both cell lines. We concluded that normal fibroblasts are more sensitive to the cytotoxic effects of the PAMAM G3 dendrimer and that modification of its surface cationic groups by substitution with biologically active molecules significantly decreases that effect, confirming that PAMAM G3 is a useful candidate as a carrier for active biocompound delivery. Keywords: ternary bioconjugate, pyridoxal, biotin, enzymatic transamination, enzymatic carboxylation, apoptosis, normal fibroblasts, squamous carcinoma cellsUram ŁSzuster MGargasz KFilipowicz AWałajtys-Rode EWołowiec SDove Medical PressarticleMedicine (General)R5-920ENInternational Journal of Nanomedicine, Vol 2013, Iss Issue 1, Pp 4707-4720 (2013)
institution DOAJ
collection DOAJ
language EN
topic Medicine (General)
R5-920
spellingShingle Medicine (General)
R5-920
Uram Ł
Szuster M
Gargasz K
Filipowicz A
Wałajtys-Rode E
Wołowiec S
In vitro cytotoxicity of the ternary PAMAM G3–pyridoxal–biotin bioconjugate
description Łukasz Uram, Magdalena Szuster, Krzysztof Gargasz, Aleksandra Filipowicz, Elżbieta Wałajtys-Rode, Stanisław Wołowiec Cosmetology Department, University of Information Technology and Management in Rzeszów, Rzeszów, Poland Abstract: A third-generation polyamidoamine dendrimer (PAMAM G3) was used as a macromolecular carrier for pyridoxal and biotin. The binary covalent bioconjugate of G3, with nine molecules of biotin per one molecule of G3 (G39B), and the ternary covalent bioconjugate of G3, with nine biotin and ten pyridoxal molecules (G39B10P), were synthesized. The biotin and pyridoxal residues of the bioconjugate were available for carboxylase and transaminase enzymes, as demonstrated in the conversion of pyruvate to oxaloacetate and alanine to pyruvate, respectively, by in vitro monitoring of the reactions, using 1H nuclear magnetic resonance spectroscopy. The toxicity of the ternary bioconjugate (BC-PAMAM) was studied in vitro on BJ human normal skin fibroblasts and human squamous cell carcinoma (SCC-15) cell cultures in comparison with PAMAM G3, using three cytotoxicity assays (XTT, neutral red, and crystal violet) and an estimation of apoptosis by confocal microscopy detection. The tests have shown that BC-PAMAM has significantly lower cytotoxicity compared with PAMAM. Nonconjugated PAMAM was not cytotoxic at concentrations up to 5 µM (NR) and 10 µM (XTT), and BC-PAMAM was not cytotoxic up to 50 µM (both assays) for both cell lines. It has been also found that normal fibroblasts were more sensitive than SCC to both PAMAM and BC-PAMAM. The effect of PAMAM and BC-PAMAM on the initiation of apoptosis (PAMAM in fibroblasts at 5 µM and BC-PAMAM at 10 µM in both cell lines) corresponded with cytotoxicity assays for both cell lines. We concluded that normal fibroblasts are more sensitive to the cytotoxic effects of the PAMAM G3 dendrimer and that modification of its surface cationic groups by substitution with biologically active molecules significantly decreases that effect, confirming that PAMAM G3 is a useful candidate as a carrier for active biocompound delivery. Keywords: ternary bioconjugate, pyridoxal, biotin, enzymatic transamination, enzymatic carboxylation, apoptosis, normal fibroblasts, squamous carcinoma cells
format article
author Uram Ł
Szuster M
Gargasz K
Filipowicz A
Wałajtys-Rode E
Wołowiec S
author_facet Uram Ł
Szuster M
Gargasz K
Filipowicz A
Wałajtys-Rode E
Wołowiec S
author_sort Uram Ł
title In vitro cytotoxicity of the ternary PAMAM G3–pyridoxal–biotin bioconjugate
title_short In vitro cytotoxicity of the ternary PAMAM G3–pyridoxal–biotin bioconjugate
title_full In vitro cytotoxicity of the ternary PAMAM G3–pyridoxal–biotin bioconjugate
title_fullStr In vitro cytotoxicity of the ternary PAMAM G3–pyridoxal–biotin bioconjugate
title_full_unstemmed In vitro cytotoxicity of the ternary PAMAM G3–pyridoxal–biotin bioconjugate
title_sort in vitro cytotoxicity of the ternary pamam g3–pyridoxal–biotin bioconjugate
publisher Dove Medical Press
publishDate 2013
url https://doaj.org/article/0eaea7dcd7e049578554a9b1654d7819
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AT wołowiecs invitrocytotoxicityoftheternarypamamg3ndashpyridoxalndashbiotinbioconjugate
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