A new approach to transfect NF-κB decoy oligodeoxynucleotides into the periodontal tissue using the ultrasound-microbubble method.
A new approach to transfect NF-κB decoy oligodeoxynucleotides into the periodontal tissue using the ultrasound-microbubble method.
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使用超声 - 微泡法转染NF-κB诱饵寡脱氧核苷酸的新方法。
DOI：
10.1038/ijos.2017.10
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发表时间：
2017-06
影响因子：
14.9
通讯作者：
Ono T
中科院分区：
文献类型：
--
作者：
Yamaguchi H;Ishida Y;Hosomichi J;Suzuki JI;Usumi-Fujita R;Shimizu Y;Kaneko S;Ono T
本研究旨在探讨超声微泡技术对核因子κ B（NF-κB）诱饵寡核苷酸（decoy oligodeoxynucleotide，ODN）转染小鼠牙龈组织的影响。通过低（LUM-Sc）和高（HUM-Sc）强度的超声照射将6-FAM标记的混杂诱饵ODN与微泡一起应用于8周龄雄性C57 BL/6 J小鼠的牙周组织，以优化超声-微泡法的转染条件。转染后2 h进行组织学检查，以比较表达与假手术组无超声辐射（A-Sc）。采用高强度超声-微泡（HUM-NF）技术将NF-κB诱骗物转染牙周组织，观察诱骗物ODN的抗炎作用。Western blot检测HUM-Sc组、HUM-NF组和对照组牙龈组织中白细胞介素（IL）-1β、IL-6和细胞间粘附分子-1（ICAM-1）的表达。荧光显微镜观察结果显示，LUM-Sc组和HUM-Sc组牙周组织中的荧光强度明显高于A-Sc组和对照组。HUM-Sc组的荧光强度最高，尤其是牙龈结缔组织。Western blot分析显示HUM-NF组IL-1β、IL-6和ICAM-1蛋白表达水平明显低于HUM-Sc组和对照组。这些结果表明，高强度超声微泡技术是一种有效的工具，诱骗转染到牙周组织。
The objective of this study is to investigate the effect of the ultrasound-microbubble technique in nuclear factor kappa B (NF-κB) decoy oligodeoxynucleotide (ODN) transfection in the gingival tissue in mice. The 6-FAM-labeled scrambled decoy ODN with microbubbles was applied to the periodontal tissue in 8-week-old male C57BL/6J mice by ultrasound radiation at low (LUM-Sc) and high (HUM-Sc) intensities to optimize the transfection condition of the ultrasound-microbubble method. Histological inspections were performed two hours after transfection to compare the expression with that in the sham-operated group without ultrasound radiation (A-Sc). Then, an NF-κB decoy was transfected into the periodontal tissue using the high-intensity ultrasound-microbubble (HUM-NF) technique to examine the anti-inflammatory effects of the decoy ODN. Western blot analysis was performed to investigate the expression of interleukin(IL)-1β, IL-6 and intercellular adhesion molecule-1 (ICAM-1) in the gingival tissues in the HUM-Sc, the HUM-NF and control groups. The fluorescence microscopy results showed that the fluorescent intensity in the periodontal tissues in the LUM-Sc and HUM-Sc groups was significantly higher than that in the A-Sc and the control groups. The fluorescent intensity in the HUM-Sc group, especially in the gingival connective tissue, was the highest of all groups. Western blot analysis indicated that the protein expression levels of IL-1β, IL-6 and ICAM-1 in the HUM-NF group were significantly lower than those in the HUM-Sc and the control groups. These findings suggest that the high-intensity ultrasound-microbubble technique is an effective tool for decoy transfection into the periodontal tissue.
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