Photodymic Therapy for Intimal Hyperplasia : Targeted Drug Delivery by Scavenger Receptors

内膜增生的光动力疗法：清道夫受体的靶向药物输送

• 批准号: 07671336
• 负责人: NAGAE Tsuneyuki
• 金额: $ 1.41万
• 依托单位: Tokyo Medical College
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07671336/
• 关键词: Intimal Hyperplasla; Photodynamic Therapy; Scavenger Receptor; Macrophage; 新生内膜胞厚

We have previously shown that the fluorescent probe maleylated-bovine serum albumin (mal-BSA) conjugated with Texas-Red can be selectively targeted to the injured sites of arteries/IH via a scavenger receptor mediated mechanism. Therefore we conjugate photosensitizer (Chlorin e6) to maleylated-BSA for selective localization. In vivo study, endothelial cells, smooth muscle cells and macrophages in culture were analyzed for their uptake of Cle6-mal-BSA conjugates. Macrophages were found to concentrate larger amount of the conjugates than either smooth muscle cells or endothelial cells. A rat model was used to evaluate fluorophore uptake in abdominal (balloon injured) and thoracic (control) arteries for three forms of Cle6 : (1)free Cle6, (2)BSA conjugated to Cle6 and(3)mal-BSA conjugated to Cle6. Fluorescent compounds were injected 2 weeks after surgery. Four hours following injection, injured and control aorta were retrieved and examined by fluorescence microscopy. Mal-BSA/Cle6 injected group showed high fluorescence distribution in the inner side of the neointima. Free Cle6 and BSA-Cle6 did not demonstrate such high fluorescence levels. Laser irradiations were perfomed with a argon pumped dye laser tuned to 661nm. We employed two energy density : 20 J/cm2 and 40 J/cm2. Forty-eight hours after PDT,the arteries were excised and examined histologically. Intimal hyperplastic cells were significantly damaged in the mal-BSA/Cle6 injected group (20J,67%) versus Cle6 group (20J,0-20%). We conclude that mal-BSA/Cle6 localizes higher inner side of IH and functions effectively as a photosensitizer for PDT.PDT as a treatment of IH using this delivery system shows effective in this model of arterial wall injury.

我们先前已经证明，与德州红偶联的马来化牛血清白蛋白(MAL-BSA)荧光探针可以通过清道夫受体介导的机制选择性地靶向动脉/IH的损伤部位。因此，我们将光敏剂(氯化e6)偶联到顺丁烯二酸化的牛血清白蛋白上进行选择性定位。在体内实验中，分析培养的内皮细胞、平滑肌细胞和巨噬细胞对Cle6-Mal-BSA结合物的摄取。巨噬细胞比血管内皮细胞或血管内皮细胞浓缩更多的偶联物。用大鼠模型评价了三种形式的Cle6：(1)游离Cle6，(2)与Cle6结合的BSA和(3)与Cle6结合的Mal-BSA对腹主动脉(球囊损伤)和胸主动脉(对照)的荧光团摄取。术后2周注射荧光化合物。注射后4h取材，荧光显微镜下观察损伤动脉和对照动脉。MAL-BSA/Cle6注射组新生内膜内侧有较强的荧光分布。游离Cle6和BSA-Cle6没有表现出如此高的荧光水平。激光照射采用调谐到661 nm的Ar泵浦染料激光器。我们采用两种能量密度：20J/cm2和40J/cm2。PDT后48小时，切除动脉并进行组织学检查。Mal-BSA/Cle6注射组(20J，67%)较Cle6组(20J，0~20%)内膜增生细胞明显受损。结论：Mal-BSA/Cle6定位于IH较高的内侧，可作为PDT的光敏剂，PDT作为治疗IH的药物，在动脉壁损伤模型中显示出良好的疗效。