N-Acetyl-l-cysteine enhances ex-vivo amplification of deciduous teeth dental pulp stem cells.
OBJECTIVE
Obtaining high number of stem cells is of interest for cell based therapies. N-Acetyl-l-cysteine (NAC) acts as a source of sulfhydryl groups and an anti-oxidative agent. The aim of this study was to test different NAC concentration on proliferation and differentiation of deciduous teeth dental pulp stem cells (DTSCs) in vitro as well as to define the possible underlining mechanism of its effect.


METHODS
Number of viable, apoptotic and senescent DTSCs was determined after addition of NAC (0.1mM, 1.0mM, 2.0mM). Also, cell cycle analysis, HIF1-α expression, LDH isoenzymes, superoxide-dismutase (SOD) and catalase (CAT) activity, sulfhydryl groups content, the level of lipids' and proteins' oxidative damage and differentiation capacity of NAC treated DTSCs was determined.


RESULTS
DTSCs expressed HIF-1α in all conditions. The lowest NAC dose (0.1mM) increased the number of DTSCs by one fifth comparing to the control, most likely stimulating entry of cells into S phase of cell cycle and enhancing the activity of LDH5 isoenzyme. The highest NAC dose (2mM) inhibited DTSCs proliferation. Also, DTSCs had the lowest level of oxidative damage with 0.1mM NAC. All tested NAC concentrations enhanced DTSCs osteo-chondrogenesis.


CONCLUSIONS
The lowest NAC dose exerted significant positive effect on DTSCs proliferation as well as antioxidative protection creating beneficial environment for stem cells in vitro cultivation especially when their clinical use is important for stimulation of osteo-chondrogenesis.
Institute for Medical Research, University of Belgrade, Dr Subotica 4, 11000 Belgrade, Serbia. Electronic address: minaizdravko@yahoo.com.
Jasmina
J
Debeljak Martacic
Faculty of Veterinary Medicine, University of Belgrade, Bulevar oslobodjenja 18, 11000 Belgrade, Serbia. Electronic address: sborozan@vet.bg.ac.rs.
Suncica
S
Borozan
Faculty of Veterinary Medicine, University of Belgrade, Bulevar oslobodjenja 18, 11000 Belgrade, Serbia. Electronic address: anita@vet.bg.ac.rs.
Anita
A
Radovanovic
Faculty of Medicine, University of Belgrade, Dr Subotica 8, 11000 Belgrade, Serbia. Electronic address: dpopadic@med.bg.ac.rs.
Dusan
D
Popadic
Institute for Medical Research, University of Belgrade, Dr Subotica 4, 11000 Belgrade, Serbia. Electronic address: slavko@imi.bg.ac.rs.
Slavko
S
Mojsilovic
Institute for Medical Research, University of Belgrade, Dr Subotica 4, 11000 Belgrade, Serbia. Electronic address: vesna.vucic.imr@gmail.com.
Vesna
V
Vucic
Faculty of Stomatology, University of Business Academy, Zarka Zrenjanina 179, 26000 Pancevo, Serbia. Electronic address: drvera.todorovic@gmail.com.
Vera
V
Todorovic
Faculty of Veterinary Medicine, University of Belgrade, Bulevar oslobodjenja 18, 11000 Belgrade, Serbia. Electronic address: milkovac@yahoo.com.
Milica
M
Kovacevic Filipovic
D000111:Acetylcysteine / Q000494:pharmacology*; D000975:Antioxidants / Q000494:pharmacology; D002374:Catalase / Q000378:metabolism; D002478:Cells, Cultured; D002648:Child; D003782:Dental Pulp / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism; D004789:Enzyme Activation; D006801:Humans; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000096:biosynthesis / Q000378:metabolism; D007527:Isoenzymes / Q000378:metabolism; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D000080864:Lactate Dehydrogenase 5; D015227:Lipid Peroxidation; D010012:Osteogenesis / Q000187:drug effects; D010084:Oxidation-Reduction; D013234:Stem Cells / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism; D013438:Sulfhydryl Compounds / Q000494:pharmacology; D013482:Superoxide Dismutase / Q000378:metabolism; D014094:Tooth, Deciduous / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism; D001616:beta-Galactosidase / Q000378:metabolism
D000975:Antioxidants; C497442:HIF1A protein, human; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D007527:Isoenzymes; D013438:Sulfhydryl Compounds; D007770:L-Lactate Dehydrogenase; D000080864:Lactate Dehydrogenase 5; D002374:Catalase; D013482:Superoxide Dismutase; D001616:beta-Galactosidase; D000111:Acetylcysteine
Antioxidative defense; Dental tissue stem cells; N-Acetyl-l-cysteine; Proliferation