| The Effects of the Oxidative Stress on the Differentiation of
Human Bone Marrow Stromal Cell to Osteoblast
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| 산화 스트레스가 골수기질세포로부터 조골세포로
분화하는 과정에 미치는 영향
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| 조정윤*,심선미*,태현정,백기현,김혜수,한제호,강무일,이광우,손호영,강성구 |
| 가톨릭대학교 의과대학 의과학 연구원*, 내과학교실 |
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| Abstract |
Background
Human bone marrow stromal cells (BMSCs) are belived to play a major role in bone formation source of osteogenitor cells. In osteoporosis, bone loss involves both increased osteoclastic bone resorption and decreased osteoblastic bone formation. Osteoporosis may be associated with increased oxidative cellular stress of free radicals in the progress of aging. Reactive Oxygen Species (ROS) enhance the osteoclastic activity, but the effect of ROS on osteoblastic function is unkown. Oxidative stress shortened the cellular replicative life span mediated by the erosion of telomeres. Telomere length decrease regularly with cell division and short telomeres induce the arrest in replicative senescence. We analyzed how oxidative stress effect the telomere length on the differentiation of human bone marrow stromal cells in ex vivo culture.
Method: Bone marrow was harvested from iliac crest. Mononuclear cells were separated using Ficoll-Hypaque, seeded in culture flasks including α-MEM at a density of 4×105 cells/ml, and incubated in a humidified atmosphere at 37℃ with 5% CO2. We treated with hydrogen peroxide (H2O2), Xanthine/Xanthine Oxidase (X/XO) and assessed their effects on intracellular oxidative stress, cell viability and telomere length. Intracellular oxidative stress was measured by 2' 7'-Dichlorofluorescin diacetate (DCF) and cell viability was measured by MTT assay. Telomere length was assessed by southern blot analysis during secondary culture.
Result: Oxidative stress (H2O2 and X/XO) increased intracellular oxidative stress as determined by DCF. Treatment with oxidative stress causes a decrease in the cell viability by dose and time dependently. But, present study did not detect any significant change with the addition of antioxidant, and telomere length was not shortened by oxidative stress.
Conclusion
The present study suggests that oxidative stress decreased cell viability. Induction of Reactive Oxygen Species (ROS) generation influence |
| Key Words:
Human bone marrow stromal cell, Reactive Oxygen Species, Telomere length |
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