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  • Hydrogen peroxide generation and biocompatibility of hydrogel-bound mussel adhesive moiety.

Hydrogen peroxide generation and biocompatibility of hydrogel-bound mussel adhesive moiety.

Acta biomaterialia (2015-02-14)
Hao Meng, Yuting Li, Madeline Faust, Shari Konst, Bruce P Lee
ABSTRACT

To decouple the extracellular oxidative toxicity of catechol adhesive moiety from its intracellular non-oxidative toxicity, dopamine was chemically bound to a non-degradable polyacrylamide hydrogel through photo-initiated polymerization of dopamine methacrylamide (DMA) with acrylamide monomers. Network-bound dopamine released cytotoxic levels of H2O2 when its catechol side chain oxidized to quinone. Introduction of catalase at a concentration as low as 7.5 U/mL counteracted the cytotoxic effect of H2O2 and enhanced the viability and proliferation rate of fibroblasts. These results indicated that H2O2 generation is one of the main contributors to the cytotoxicity of dopamine in culture. Additionally, catalase is a potentially useful supplement to suppress the elevated oxidative stress found in typical culture conditions and can more accurately evaluate the biocompatibility of mussel-mimetic biomaterials. The release of H2O2 also induced a higher foreign body reaction to catechol-modified hydrogel when it was implanted subcutaneously in rat. Given that H2O2 has a multitude of biological effects, both beneficiary and deleterious, regulation of H2O2 production from catechol-containing biomaterials is necessary to optimize the performance of these materials for a desired application.

MATERIALS
Product Number
Brand
Product Description

Supelco
Sorbitol, Pharmaceutical Secondary Standard; Certified Reference Material
Supelco
Dehydrated Alcohol, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
N,N′-Methylenebis(acrylamide), 99%
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D-Sorbitol, 99% (GC)
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D-Sorbitol, liquid, tested according to Ph. Eur.
Supelco
Acrylamide, analytical standard
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Ethanol solution, certified reference material, 2000 μg/mL in methanol
Millipore
Hydrogen peroxide solution, 3%, suitable for microbiology
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D-Sorbitol, FCC, FG
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Sodium pyruvate, BioXtra, ≥99%
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Sodium pyruvate, powder, BioReagent, suitable for cell culture, suitable for insect cell culture, ≥99%
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Sodium pyruvate, ReagentPlus®, ≥99%
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Acrylamide, suitable for electrophoresis, ≥99% (HPLC), powder
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N,N′-Methylenebisacrylamide, powder, Molecular Biology, suitable for electrophoresis, ≥99.5%
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D-Sorbitol, ≥98% (GC)
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Acrylamide, suitable for electrophoresis, ≥99%
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D-Sorbitol, ≥98% (GC), Molecular Biology
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Ethanol standards 10% (v/v), 10 % (v/v) in H2O, analytical standard
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Dopamine hydrochloride
USP
Dehydrated Alcohol, United States Pharmacopeia (USP) Reference Standard
Dopamine hydrochloride, European Pharmacopoeia (EP) Reference Standard
Sorbitol, European Pharmacopoeia (EP) Reference Standard
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Hydrogen Peroxide Solution, 30% (w/w), puriss. p.a., reag. ISO, reag. Ph. Eur.
USP
Sorbitol, United States Pharmacopeia (USP) Reference Standard
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Ethyl alcohol, Pure 190 proof, for molecular biology
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Hydrogen peroxide solution, contains inhibitor, 30 wt. % in H2O, ACS reagent
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Hydrogen peroxide solution, contains ~200 ppm acetanilide as stabilizer, 3 wt. % in H2O
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Acrylamide, certified reference material, TraceCERT®, Manufactured by: Sigma-Aldrich Production GmbH, Switzerland
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Ethyl alcohol, Pure, 200 proof, anhydrous, ≥99.5%
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Dimethyl sulfoxide, sterile-filtered, BioPerformance Certified, meets EP, USP testing specifications, suitable for hybridoma