Synthesis, Swelling Properties and Evaluation of Genotoxicity of Hydrogels Based on (Meth)acrylates and Itaconic Acid

Takić Miladinov, Dijana; Tomić, Simonida; Stojanović, Sanja; Najdanović, Jelena; Filipović, Jovanka; Trajanović, Miroslav; Najman, Stevo

doi:10.1590/1980-5373-MR-2016-0222

Dijana Takić Miladinov ^** e-mail: takicdijana@gmail.com

Department of Biology and Ecology, Faculty of Sciences and Mathematics, University of Niš, Niš, Serbia

Simonida Tomić

Department of Organic Chemical Technology, Faculty of Technology and Metallurgy, University of Belgrade, Belgrade, Serbia

Sanja Stojanović

Department for Cell and Tissue Engineering, Faculty of Medicine, University of Niš, Niš, Serbia

Department of Biology and Human Genetics, Faculty of Medicine, University of Niš, Niš, Serbia

Jelena Najdanović

Department for Cell and Tissue Engineering, Faculty of Medicine, University of Niš, Niš, Serbia

Department of Biology and Human Genetics, Faculty of Medicine, University of Niš, Niš, Serbia

Jovanka Filipović

Department of Organic Chemical Technology, Faculty of Technology and Metallurgy, University of Belgrade, Belgrade, Serbia

Miroslav Trajanović

Laboratory for Intelligent Production Systems, Faculty of Mechanical Engineering, University of Niš, Niš, Serbia

Stevo Najman

Department for Cell and Tissue Engineering, Faculty of Medicine, University of Niš, Niš, Serbia

Department of Biology and Human Genetics, Faculty of Medicine, University of Niš, Niš, Serbia

* e-mail: takicdijana@gmail.com

6. Conflict of interests

Authors declare that they have no conflict of interests.

Monomer	Cell line	Genotoxicity test (monomer concentration)	Result	Reference
HEMA	V79 Chinese hamster lung fibroblasts	Micronucleus Comet assay (1-18 mM)	Genotoxic effect in a dose-dependent manner.	Lee et al. 2006²⁸29 Lee DH, Lim BS, Lee YK, Ahn SJ, Yang HC. Involvement of oxidative stress in mutagenicity and apoptosis caused by dental resin monomers in cell cultures. Dental Materials. 2006;22(12):1086-1092.
	V79 Chinese hamster lung fibroblasts	Micronucleus (2-8 mM)	Genotoxic effect in a dose-dependent manner.	Schweikl et al. 2007²⁹29 Schweikl H, Hartmann A, Hiller KA, Spagnuolo G, Bolay C, Brockhoff G, Schmalz G. Inhibition of TEGDMA and HEMA-induced genotoxicity and cell cycle arrest by N-acetylcysteine. Dental Materials. 2007;23(6):688-695.
	Human peripheral blood lymphocytes	Comet assay	Mild enhancement of DNA migration.	Kleinsasser et al. 2004³⁰30 Kleinsasser NH, Wallner BC, Harréus UA, Kleinjung T, Folwaczny M, Hickel R, et al. Genotoxicity and cytotoxicity of dental materials in human lymphocytes as assessed by the single cell microgel electrophoresis (comet) assay. Journal of Dentistry. 2004;32(3):229-234.
	Human samples of salivary glands Human peripheral blood lymphocytes	Comet assay	Significant DNA migration in both cell types.	Kleinsasser et al. 2006³¹31 Kleinsasser NH, Schmid K, Sassen AW, Harreus UA, Staudenmajer R, Folwaczny M, et al. Cytotoxic and genotoxic effects of resin monomers in human salivary gland tissue and lymphocytes as assessed by the single cell microgel electrophoresis (comet) assay. Biomaterials. 2006;27(9):1762-1770.
	Human peripheral blood lymphocytes A549 lung-tumour cells	Comet assay (0-10 mM)	Increased DNA damage in a dose-dependent manner	Pawlowska et al. 2010³²32 Pawlowska E, Poplawski T, Ksiazek D, Szczepanska J, Blasiak J. Genotoxicity and cytotoxicity of 2-hydroxyethyl methacrylate. Mutation Research. 2010;696(2):122-129.
	Human peripheral blood lymphocytes	Comet assay Micronucleus Chromosome aberration (CA) Sister chromatid exchange (SCE) (10 µM-1 mM)	Genotoxic effect was measurable in the comet assay at 1 mM of HEMA, but not in the micronucleus test. A significant dose-dependent increase in the frequency of CAs and SCEs could be demonstrated in all tested concentrations.	Ginzkey et al. 2015³³33 Ginzkey C, Zinnitsch S, Steussloff G, Koehler C, Hackenberg S, Hagen R, et al. Assessment of HEMA and TEGDMA induced DNA damage by multiple genotoxicological endpoints in human lymphocytes. Dental Materials. 2015;31(8):865-876.
	Human gingival fibroblasts	Comet assay (1-10 mM)	Increased tail DNA in a dose-dependent manner.	Szczepanska et al. 2012³⁴34 Szczepanska J, Poplawski T, Synowiec E, Pawlowska E, Chojnacki CJ, Chojnacki J, et al. 2-hydroxylethyl methacrylate (HEMA), a tooth restoration component, exerts its genotoxic effects in human gingival ?broblasts trough methacrylic acid, an immediate product of its degradation. Molecular Biology Reports. 2012;39(2):1561-1574.
IA	N/A
PAGM	N/A
Hydrogel	Cell line	Genotoxicity test (extract concentration)	Result	Reference
NVP-AA^a a N-vinyl-2-pyrrolidone/acrylic acid	HepG2	Comet assay (0.25-25 mg/ml)	Four to six fold increase in DNA breaks.	Devine et al. 2006⁵5 Devine DM, Devery SM, Lyons JG, Geever LM, Kennedy JE, Higginbotham CL. Multifunctional polyvinylpyrrolidinone-polyacrylic acid copolymer hydrogels for biomedical applications. International Journal of Pharmaceutics. 2006;326(1-2):50-59.
NVP-AA	HepG2 HaCaT	Comet assay Ames assay	Genocompatible	Kirf et al. 2010²³23 Kirf D, Higginbotham CL, Rowan NJ, Devery SM. Cyto- and genotoxicological assessment and functional characterization of N-vinyl-2-pyrrolidone-acrylic acid-based copolymeric hydrogels with potential for future use in wound healing applications. Biomedical Materials (Bristol, England). 2010;5(3):35002.
ALPF-HEMA^b b Alginate-polypropylene fumarate/2-hydroxyethyl methacrylate	L929 fibroblasts	Comet assay	Genocompatible	Finosh et al. 2014³⁵35 Thankam FG, Muthu J. Infiltration and sustenance of viability of cells by amphiphilic biosynthetic biodegradable hydrogels. Journal of Materials Science: Materials in Medicine. 2014;25(8):1953-1965.

Component	P(HEMA/IA/PAGM1)	P(HEMA/IA/PAGM2)	PHEMA
HEMA (mol%)	70	70	100
PAGM 1 and 2 (mol%)	28	28	0
IA (mol%)	2	2	0
HEMA+IA+PAGM (wt%)	10	10	10
Demineralized water (wt%)	45	45	45
Ethyl alcohol (wt%)	45	45	45
Conversion (wt%)	97.5	98.1	99

Treatment	Cell viability (%)	DNA damage ± S.E.M
Treatment	Cell viability (%)	Mean %DNA_t	Mean M_t
Untreated control	94.8	3.19 ± 0.63	0.41 ± 0.21
PHEMA
10%	89.7	6.19 ± 1.47	0.76 ± 0.62
50%	79.4	11.03 ± 1.39*	1.59 ± 0.53*
P(HEMA/IA/PAGM1)
10%	88.1	7.47 ± 1.8	1.53 ± 0.42
50%	76.6	25.39 ± 4.62*	8.78 ± 1.35*
P(HEMA/IA/PAGM2)
10%	87.8	6.57 ± 0.94	0.71 ± 0.26
50%	74.7	23.82 ± 2.15*	4.63 ± 0.85*
Positive control^a	69.5	21.11 ± 1.85*	8.59 ± 1.71*

\begin{matrix} \frac{V_{1} X^{2} φ_{2, s}^{2}}{4 I {\overset{─}{V}}_{r}^{2}} {(\frac{2 K_{a 1} K_{a 2} + 10^{- pH} K_{a 1}}{2 {(10^{- pH})}^{2} + 10^{- pH} K_{a 1} + K_{a 1} K_{a 2}})}^{2} = \\ [\ln (1 - φ_{2, s}) + φ_{2, s} + {χ φ}_{2, s}^{2}] + (\frac{V_{1} ρ}{{\overset{─}{M}}_{c}}) φ_{2, r}^{2 / 3} φ_{2, s}^{1 / 3} \end{matrix}

[1] * e-mail: takicdijana@gmail.com

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Synthesis, Swelling Properties and Evaluation of Genotoxicity of Hydrogels Based on (Meth)acrylates and Itaconic Acid