Jasmina Martacic1, Milica Kovacevic Filipovic2, Suncica Borozan2, Zorica Cvetkovic3,4, Tamara Popovic1, Aleksandra Arsic1, Marija Takic1, Vesna Vucic5, Maria Glibetic1. 1. Institute for Medical Research, University of Belgrade, Dr Subotica 4, Belgrade, 11000, Serbia. 2. Faculty of Veterinary Medicine, University of Belgrade, Bulevar oslobodjenja 18, Belgrade, 11000, Serbia. 3. Department of Hematology, Clinical Hospital Center Zemun, Vukova 9, Belgrade, 11080, Serbia. 4. Faculty of Medicine, University of Belgrade, Dr Subotića 8, Belgrade, 11000, Serbia. 5. Institute for Medical Research, University of Belgrade, Dr Subotica 4, Belgrade, 11000, Serbia. vesna.vucic.imr@gmail.com.
Abstract
OBJECTIVES: The aim of our study was to investigate whether N-acetyl-L-cysteine (NAC) could protect stem cells from exfoliated deciduous teeth (SHED) against oxidative damage, during in vitro cultivation, to preserve regenerative potential of these cells. Accordingly, we examined the potential of cell culture supplementation with NAC in prevention of lipid peroxidation, unfavorable changes of total lipids fatty acid composition, and the effects on the activity of antioxidant enzymes. MATERIAL AND METHODS: We analyzed the extent of oxidative damage in SHED after 48 h treatment with different NAC concentrations. Cellular lipid peroxidation was determined upon reaction with thiobarbituric acid. All enzyme activities were measured spectrophotometrically, based on published methods. Fatty acid methyl esters were analyzed by gas-liquid chromatography. RESULTS: Concentration of 0.1 mM NAC showed the most profound effects on SHED, significantly decreasing levels of lipid peroxidation in comparison to control. This dose also diminished the activities of antioxidant enzymes. Furthermore, NAC treatment significantly changed fatty acid composition of cells, reducing levels of oleic acid and monounsaturated fatty acids and increasing linoleic acid, n-6, and total polyunsaturated fatty acid (PUFA) proportions. CONCLUSION: Low dose of NAC significantly decreased lipid peroxidation and altered fatty acid composition towards increasing PUFA. The reduced oxidative damage of cellular lipids could be strongly related to improved SHED survival in vitro. CLINICAL RELEVANCE: Low doses of antioxidants, applied during stem cells culturing and maintenance, could improve cellular characteristics in vitro. This is prerequisite for successful use of stem cells in various clinical applications.
OBJECTIVES: The aim of our study was to investigate whether N-acetyl-L-cysteine (NAC) could protect stem cells from exfoliated deciduous teeth (SHED) against oxidative damage, during in vitro cultivation, to preserve regenerative potential of these cells. Accordingly, we examined the potential of cell culture supplementation with NAC in prevention of lipid peroxidation, unfavorable changes of total lipidsfatty acid composition, and the effects on the activity of antioxidant enzymes. MATERIAL AND METHODS: We analyzed the extent of oxidative damage in SHED after 48 h treatment with different NAC concentrations. Cellular lipid peroxidation was determined upon reaction with thiobarbituric acid. All enzyme activities were measured spectrophotometrically, based on published methods. Fatty acid methyl esters were analyzed by gas-liquid chromatography. RESULTS: Concentration of 0.1 mM NAC showed the most profound effects on SHED, significantly decreasing levels of lipid peroxidation in comparison to control. This dose also diminished the activities of antioxidant enzymes. Furthermore, NAC treatment significantly changed fatty acid composition of cells, reducing levels of oleic acid and monounsaturated fatty acids and increasing linoleic acid, n-6, and total polyunsaturated fatty acid (PUFA) proportions. CONCLUSION: Low dose of NAC significantly decreased lipid peroxidation and altered fatty acid composition towards increasing PUFA. The reduced oxidative damage of cellular lipids could be strongly related to improved SHED survival in vitro. CLINICAL RELEVANCE: Low doses of antioxidants, applied during stem cells culturing and maintenance, could improve cellular characteristics in vitro. This is prerequisite for successful use of stem cells in various clinical applications.