Purpose To assess the maximum uptake of Iododeo xyur idine (IUdR) by proliferating smooth muscle cells in vitro to determine the opti mal concentration to be administrated in an in vivo experiment. The long-term g oal...Purpose To assess the maximum uptake of Iododeo xyur idine (IUdR) by proliferating smooth muscle cells in vitro to determine the opti mal concentration to be administrated in an in vivo experiment. The long-term g oal is to utilize radioactive IUdR to inhibit smooth muscle cell proliferation a nd restenosis of arteries after balloon angioplasty in vivo. Methods Porcine smooth muscle cells (SMCs) were cultured in 5% FBS medium and stim ulated to proliferate by the addition of medium containing 10% FBS and insulin. IUdR was added at 5 μM, 10 μM, 20 μM, 30 μM, 40 μM, respectively, in prolif erating SMCs with control for 1, 3, 5, 7 day incubation. Fluorescence Activated Cell Scanning (FACS) was performed after the SMCs were harvested and double-sta ined with FITC-conjugated anti-IUdR antibody (B44) and propidium iodide (PI). The ratio of IUdR-labeled cells to total cell population for each IUdR concentr ation and duration was determined by FACS. All data were repeated three times at each time point. The doubling times, growth curve and cell density of the proli ferating SMCs were investigated using Beckman Coulter Particle Counter and digit al microscopy. Results The percentage of proliferating SMCs uptaking IUdR incr eased from 1 to 5 days incubation with all concentrations of IUdR; In day 5, the uptake rate reached the peak value, then decreased by 7 days. IUdR uptake on d ay 5 was higher with concentrations of 10 μM and 20 μM. The doubling times of the SMCs were prolonged with IUdR concentration increasing, while the proliferat ing cell number and density compared with control decreased obviously by day 5 ( P<0.05).Conclusion The peak time to uptake IUdR was 5 days and optimal concentration of IUdR was between10 μM to 20 μM for proliferating SMCs to upta ke in vitro. IUdR itself could inhibit the SMCs’ proliferation and the inhibito ry effect was related to the concentration.[展开更多
Objective To investigate the kinetics of Iododeoxyuridine(IUdR)release from sodium alginate hydrogel cross-linked with varying amounts of calcium chloride, and to optimize sustained release for further periadventitial...Objective To investigate the kinetics of Iododeoxyuridine(IUdR)release from sodium alginate hydrogel cross-linked with varying amounts of calcium chloride, and to optimize sustained release for further periadventitial I125-labeled IUdR delivery to suppress intimal hyperplasia following angioplasty in vivo. Methods Four hydrogels,composed of 0.16 mEq sodium alginate and 200 g IUdR, were cross-linked with calcium chloride to yield ion equivalence (IE) ratios (Calcium: alginate) of 3:1, 4:1, 5:1, or 6:1. 2 ml of normal saline was placed on top of each hydrogel and allowed to remain in contact at 37℃ for up to 30 days. At set time intervals, the concentration and amount of IUdR in the eluate were assayed by high performance liquid chromatography using UV detection and Water symmetry C18 column. The data for accumulated release rate and concentration in the eluate were calculated based on the calibration curve of peak area versus IUdR concentration. The hydrogel morphologic degradations were also observed. Results The hydrogels entrapped 92.9%, 98.6%, 98.4% and 98.6% of the IUdR with 3:1, 4:1, 5:1 and 6:1 IE ratios, respectively. IUdR concentration in eluates from 3:1 IE ratio hydrogel decreased faster than that from other hydrogels over time (P < 0.01). The 4:1, 5:1 and 6:1 IE ratio hydrogels produced more than 10 μm IUdR concentrations in eluates for the first 8 days, while the 3:1 IE ratio hydrogel for 4 days. IUdR release rates of the 4:1, 5:1 and 6:1 IE ratio hydrogels were very close, however they were lower than that of the 3:1 IE hydrogel in the first 48 hours (P < 0.05). At day 30, the 3:1 and 4:1 IE ratio hydrogels had 100% and 88% degradation, but no significant degradation was observed in the other hydrogels. Conclusion The sodium alginate hydrogel with 4:1 IE ratio exhibited an optimal IUdR sustained release and almost complete degradation in 30 days. (J Intervent Radiol, 2006, 15: 293-298)展开更多
文摘Purpose To assess the maximum uptake of Iododeo xyur idine (IUdR) by proliferating smooth muscle cells in vitro to determine the opti mal concentration to be administrated in an in vivo experiment. The long-term g oal is to utilize radioactive IUdR to inhibit smooth muscle cell proliferation a nd restenosis of arteries after balloon angioplasty in vivo. Methods Porcine smooth muscle cells (SMCs) were cultured in 5% FBS medium and stim ulated to proliferate by the addition of medium containing 10% FBS and insulin. IUdR was added at 5 μM, 10 μM, 20 μM, 30 μM, 40 μM, respectively, in prolif erating SMCs with control for 1, 3, 5, 7 day incubation. Fluorescence Activated Cell Scanning (FACS) was performed after the SMCs were harvested and double-sta ined with FITC-conjugated anti-IUdR antibody (B44) and propidium iodide (PI). The ratio of IUdR-labeled cells to total cell population for each IUdR concentr ation and duration was determined by FACS. All data were repeated three times at each time point. The doubling times, growth curve and cell density of the proli ferating SMCs were investigated using Beckman Coulter Particle Counter and digit al microscopy. Results The percentage of proliferating SMCs uptaking IUdR incr eased from 1 to 5 days incubation with all concentrations of IUdR; In day 5, the uptake rate reached the peak value, then decreased by 7 days. IUdR uptake on d ay 5 was higher with concentrations of 10 μM and 20 μM. The doubling times of the SMCs were prolonged with IUdR concentration increasing, while the proliferat ing cell number and density compared with control decreased obviously by day 5 ( P<0.05).Conclusion The peak time to uptake IUdR was 5 days and optimal concentration of IUdR was between10 μM to 20 μM for proliferating SMCs to upta ke in vitro. IUdR itself could inhibit the SMCs’ proliferation and the inhibito ry effect was related to the concentration.[
文摘Objective To investigate the kinetics of Iododeoxyuridine(IUdR)release from sodium alginate hydrogel cross-linked with varying amounts of calcium chloride, and to optimize sustained release for further periadventitial I125-labeled IUdR delivery to suppress intimal hyperplasia following angioplasty in vivo. Methods Four hydrogels,composed of 0.16 mEq sodium alginate and 200 g IUdR, were cross-linked with calcium chloride to yield ion equivalence (IE) ratios (Calcium: alginate) of 3:1, 4:1, 5:1, or 6:1. 2 ml of normal saline was placed on top of each hydrogel and allowed to remain in contact at 37℃ for up to 30 days. At set time intervals, the concentration and amount of IUdR in the eluate were assayed by high performance liquid chromatography using UV detection and Water symmetry C18 column. The data for accumulated release rate and concentration in the eluate were calculated based on the calibration curve of peak area versus IUdR concentration. The hydrogel morphologic degradations were also observed. Results The hydrogels entrapped 92.9%, 98.6%, 98.4% and 98.6% of the IUdR with 3:1, 4:1, 5:1 and 6:1 IE ratios, respectively. IUdR concentration in eluates from 3:1 IE ratio hydrogel decreased faster than that from other hydrogels over time (P < 0.01). The 4:1, 5:1 and 6:1 IE ratio hydrogels produced more than 10 μm IUdR concentrations in eluates for the first 8 days, while the 3:1 IE ratio hydrogel for 4 days. IUdR release rates of the 4:1, 5:1 and 6:1 IE ratio hydrogels were very close, however they were lower than that of the 3:1 IE hydrogel in the first 48 hours (P < 0.05). At day 30, the 3:1 and 4:1 IE ratio hydrogels had 100% and 88% degradation, but no significant degradation was observed in the other hydrogels. Conclusion The sodium alginate hydrogel with 4:1 IE ratio exhibited an optimal IUdR sustained release and almost complete degradation in 30 days. (J Intervent Radiol, 2006, 15: 293-298)
