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Netilmicin-induced modulation of melanogenesis in HEMa-LP melanocytes.

Acta poloniae pharmaceutica (2013-10-24)
Dorota Wrześniok, Artur Beberok, Michał Otreba, Ewa Buszman
ABSTRACT

It is known that aminoglycoside antibiotics bind well to melanin biopolymer, but the relation between their affinity to melanin and ototoxicity is not well established. The aim of this work was to examine the impact of netilmicin on melanogenesis in cultured normal human melanocytes (HEMa-LP). The WST-1 assay was used to detect netilmicin cytotoxic effect. Netilmicin induced concentration-dependent loss in melanocytes viability. The value of EC50 was found to be - 75.0 mM. The analyzed antibiotic inhibited melanin biosynthesis in a concentration-dependent manner. Increasing the drug concentration resulted also in a decrease of cellular tyrosinase activity. Netilmicin-induced modulation of melanogenesis in melanocytes in vitro may explain a potential role of melanin and melanocytes in the mechanisms of aminoglycosides ototoxic effects in vivo.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Netilmicin sulfate salt
Sigma-Aldrich
Tyrosinase from mushroom, lyophilized powder, ≥1000 unit/mg solid
USP
Levodopa, United States Pharmacopeia (USP) Reference Standard
Supelco
Levodopa, Pharmaceutical Secondary Standard; Certified Reference Material
Levodopa, European Pharmacopoeia (EP) Reference Standard
Sigma-Aldrich
Ethylenediaminetetraacetic acid solution, 0.02% in DPBS (0.5 mM), sterile-filtered, BioReagent, suitable for cell culture
Sigma-Aldrich
Ethylenediaminetetraacetic acid, 99.995% trace metals basis
Sigma-Aldrich
Ethylenediaminetetraacetic acid, ACS reagent, 99.4-100.6%, powder
Sigma-Aldrich
Ethylenediaminetetraacetic acid, ≥98.0% (KT)
Sigma-Aldrich
Ethylenediaminetetraacetic acid, BioUltra, anhydrous, ≥99% (titration)
Sigma-Aldrich
Ethylenediaminetetraacetic acid disodium salt solution, BioUltra, pH 8.0, ~0.5 M in H2O
Sigma-Aldrich
Ethylenediaminetetraacetic acid, anhydrous, crystalline, BioReagent, suitable for cell culture
Sigma-Aldrich
Ethylenediaminetetraacetic acid, purified grade, ≥98.5%, powder
Sigma-Aldrich
Ethylenediaminetetraacetic acid, BioUltra, ≥99.0% (KT)
Sigma-Aldrich
3,4-Dihydroxy-L-phenylalanine, ≥98% (TLC)
Sigma-Aldrich
Ethylenediaminetetraacetic acid, Vetec, reagent grade, 98%
Sigma-Aldrich
3,4-Dihydroxy-L-phenylalanine, Vetec, reagent grade, 98%