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The direct action of hyaluronic acid on human U-937 and HL-60 cells – modification of native and model membranes

  • Anna Barbasz , Barbara Kreczmer , Barbara Dyba , Maria Filek and Elżbieta Rudolphi-Skórska EMAIL logo
Published/Copyright: December 25, 2016
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Abstract

The aim of the study was to investigate the effect of application of exogenous hyaluronic acid (HA) of various molecular weights on the cells of human immune system. Two cell lines HL-60 and U-937 with various ability to differentiation were chosen. HL-60 cells were differentiated to macrophages and granulocytes, whereas U-937 only to macrophages. For all investigated cell systems the most cytotoxic effect, indicated as a decrease of cell viability, was found at HA dose equal to 200 mg/L. However, greater effect was observed for differentiated cells and at longer exposure to HA. The possibility of HA interaction with both specific receptors and membrane lipids was tested by determination of biotin-labelled HA binding to cell surface and analysis of physicochemical parameters of model membranes.

Abbreviations

A

area per lipid molecule

Alim

limiting area per molecule representing maximal density of a layer being characteristic for each lipid

Cs1

static compression modulus

DMSO

dimethyl sulfoxide

DOPA

1,2-sn-glycero-3-phosphate 18:1

DOPC

1,2-dioleoyl-sn-glycero-3-phosphocholine 18:1

DOPG

dioleoyl-sn-glycero-3-phospho-(1’-rac-glycerol) 18:1

DPPC

1,2-dipalmitoyl-sn-glycero-3-phosphocholine 16:0

ECM

extracellular matrix

ELLSA

enzyme-linked ligand sorbent assay

FBS

foetal bovine serum

GAG

glycosaminoglycans

HA

sodium hyaluronate, hyaluronic acid

HA-Bt

hyaluronan biotin sodium salt

HALMW

low molecular weight hyaluronic acid

iNOS

nitric oxide synthase inhibitor

MTT tetrazolium salt

methylthiazolyldiphenyl-tetrazolium bromide

PA

phosphatidic acid

PBS

phosphate buffered saline

PC

phospatidylcholine

PG

phosphatidylglycerol

PMA

12-myristate-13-acetate

SA-HRP

streptavidin conjugated to horseradish peroxidase

TMB

3,3’,5,5’-tetramethylbenzidine

π

surface pressure

πcoll

pressure at which layer collapse

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Received: 2016-6-3
Accepted: 2016-11-22
Published Online: 2016-12-25
Published in Print: 2016-11-1

© 2016 Institute of Molecular Biology, Slovak Academy of Sciences

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