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Re-oxygenation Improves Hypoxia-induced Pulp Cell Arrest
Y. Ueno1,
C. Kitamura1,
M. Terashita1 and
T. Nishihara2,*
1 Division of Pulp Biology, Operative Dentistry, and Endodontics, Department of Cariology and Periodontology, Science of Oral Functions, and
2 Division of Infections and Molecular Biology, Department of Health Promotion, Science of Health Improvement, Kyushu Dental College, 2-6-1 Manazuru, Kokurakita, Kitakyushu 803-8580, Japan
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Figure 1. The viability of RPC-C2A-K4 cells exposed to hypoxia for 0 to 60 hrs was monitored by MTT assay. White bar = cells under normoxic condition; grey bar = cells under hypoxic condition. Data are expressed as the mean ± SD of triplicate cultures. The experiment was performed 3 times, with similar results obtained in each experiment. Statistical differences were determined between the cells in normoxia and the cells in hypoxia at each time-point. *p < 0.01.
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Figure 2. Cell-cycle analysis of RPC-C2A-K4 cells exposed to hypoxia. (A) Representative results of cell-cycle distribution of the cells under normoxic (non-treated) and hypoxic conditions for 24 and 48 hrs. (B) Changes that occurred during each phase of cell-cycle distribution during hypoxia are indicated at the times noted. (C) Representative morphologies of non-treated and hypoxia-exposed cells. Scale bar = 10.0 µm.
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Figure 3. Expression of cell-cycle-related molecules in hypoxia-exposed RPC-C2A-K4 cells. The immunoblot analysis was carried out as described in MATERIALS & METHODS. pRb, hypophosphorylated Rb; ppRb, hyperphosphorylated Rb; pp53, phosphorylated p53.
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Figure 4. Improvement in intracellular functions of hypoxia-exposed RPC-C2A-K4 cells by re-oxygenation. After the cells were exposed to hypoxia for 24 hrs, cells were incubated under a normoxic condition (re-oxygenation). (A) Cell viability was monitored by MTT assay. Data are expressed as the mean ± SD of triplicate cultures. The experiment was performed 3 times, with similar results obtained in each experiment. Statistical differences were determined between the cells exposed to 24 hrs of hypoxia (0 hr after re-oxygenation) and the cells exposed to normoxia after 24 hrs of hypoxia (a, 12 hrs of re-oxygenation; b, 24 hrs of re-oxygenation; c, 36 hrs of re-oxygenation). *p < 0.01. (B) DNA content was analyzed by flow cytometry. Non-treated, cells in normoxia. (C) Expression of cyclin D2 in re-oxygenated RPC-C2A-K4 cells.
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Journal of Dental Research, Vol. 85, No. 9,
824-828 (2006)
DOI: 10.1177/154405910608500909
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