Praetorius M, Pfannenstiel S, Klingmann C, Baumann I, Plinkert P K, Staecker H
Sektion Otologie und Neuro-Otologie, Hals-, Nasen-, Ohrenklinik mit Poliklinik, Universitätsklinikum Heidelberg, Im Neuenheimer Feld 400, 69120 Heidelberg.
HNO. 2008 May;56(5):524-9. doi: 10.1007/s00106-008-1738-6.
Diseases of the inner ear such as presbycusis, tinnitus, sudden hearing loss, and vertigo affect many patients, but so far there are no specific therapy options. Gene therapy might become a potential modality of treatment. Viral vectors are standard in animal models to date. Future considerations, however, call for a further evaluation of non-viral transfection methods.
The non-viral transfection agents Metafectene, Superfect, Effectene, and Mirus TransIT were incubated with a plasmid coding for GFP. In vivo, the plasmid-agent mix was injected via the membrane of the round window, and 48 h later the inner ear was perfused, harvested, decalcified, and histologically evaluated for GFP expression.
Cationic lipids (Metafectene) and dendrimers (Superfect) were able to transfect cells in the area of the organ of Corti and lead to GFP expression. The polyamine (Mirus TransIT) did show expression of GFP in the area of Rosenthal's canal and in the area of the inner hair cell. The combination of a non-liposomal lipid with a DNA condensing component (Effectene) did not show transfection of the organ of Corti. In the area of the spiral ganglia cells, GFP expression was seen with all the transfection agents.
Non-viral transfection agents are able to introduce a reporter gene in cells of the inner ear in vitro and in vivo. There are, however, differences in the efficiency of the transfection. They might be an alternative in gene therapy of the inner ear. Further investigations to elucidate their potential are needed.
诸如老年性聋、耳鸣、突发性听力损失和眩晕等内耳疾病影响着众多患者，但迄今为止尚无特异性治疗方案。基因治疗可能成为一种潜在的治疗方式。病毒载体是目前动物模型中的标准方法。然而，未来的考量要求进一步评估非病毒转染方法。
将非病毒转染试剂Metafectene、Superfect、Effectene和Mirus TransIT与编码绿色荧光蛋白（GFP）的质粒一起孵育。在体内，将质粒 - 试剂混合物经圆窗膜注射，48小时后对内耳进行灌注、取材、脱钙，并进行组织学评估以检测GFP表达。
阳离子脂质（Metafectene）和树枝状聚合物（Superfect）能够转染柯蒂氏器区域的细胞并导致GFP表达。多胺（Mirus TransIT）在螺旋神经节管区域和内毛细胞区域确实显示出GFP表达。非脂质体脂质与DNA凝聚成分的组合（Effectene）未显示柯蒂氏器的转染。在螺旋神经节细胞区域，所有转染试剂均可见GFP表达。
非病毒转染试剂能够在体外和体内将报告基因导入内耳细胞。然而，转染效率存在差异。它们可能是内耳基因治疗的一种替代方法。需要进一步研究以阐明其潜力。
