Translational Research for axona regeneration by using Stroke model of minature pig

利用小型猪中风模型进行轴突再生的转化研究

基本信息

批准号：
17591499
负责人：
IMAI Hideaki
金额：
$ 2.18万
依托单位：
Gunma University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2005
资助国家：
日本
起止时间：
2005 至 2006
项目状态：
已结题

项目摘要

Background and Purpose The first purpose of the present set of studies was to introduce a reproducible stroke model in the gyrencephalic brain of miniature pig with minimum invasiveness. The paper provides information on both the surgical technique and the methods of quantification of ischemic damage to both grey and white matter in the miniature pig. The second purpose was to apply this new stroke model for the translational research for the neuronal regeneration by using thestem cell trasplantation into the ischemic brain. Methods Sixteen male miniature pigs were randomly divided into three groups and underwent transcranial surgery, a frontotemporal approach with orbital rim osteotomy, for : permanent middle cerebral artery occlusion (MCAO) (n=5), permanent internal carotid artery occlusion (ICAO) (n=6) and sham operation (n=5). Histological mapping and magnetic resonance (MR) imaging were used to delineate the areas of ischemia. Subventricular zone (SVZ) cells isolated from the mini … More ature pig were cultured in Iscove's modified Dulbecco's medium in the presence of basic fibroblast growth factor for 8 days. We stereotaxically transplanted subventricular zone (SVZ) cells labeled by ferromagnetic particles into stroked miniature pig. The transplanted cells were non-invasively tracked using magnetic resonance imaging (MRI). To detect ferromagnetic-labeled SVZ cells in the host brain, we stained sagittal or coronal sections for iron using Prussian blue reaction. Results The volumes of the infarction measured directly from MR images were 16.2 ±1.1, 1.5±0.5 and 0.0±0.0 cm^3 (mean±SD) in the MCAO, ICAO and sham-operation groups, respectively. The histological and MR imaging volumes showed a good correlation (r^2=0.86, p <0.0001) although the infarct volume measured by quantitative histopathology was smaller than that measured by MRI. Immunohistochemical staining using amyloid precursor protein antibody (APP) allowed us to evaluate the axonal damage, providing total APP score of 43.8±3.5 (mean±SD) in the MCAO. MRI signals detected the tranplanted cells with labeling at the lesion, and these signals were detected 1 and 2 weeks after transplantation. Prussian blue-positive cells were also detected in the same lesion where the cells were transplanted. The transplanted cells were likely to be just homing in the host brain. We have neither confirmed the construction of the network between the transplanted cells and host brain. Conclusions This paper describes a simple and reproducible method for the induction of white and grey matter damage in focal cerebral ischemia with minimum invasiveness. This miniature pig stroke model has utility for studying the pathophysiology of ischemia in gyrencephalic brain and for the assessing the therapeutic efficacy of drugs or stem cell transplants, prior to initiating human clinical trials. Less

背景和目的本集研究的第一个目的是在微型猪的小脑脑大脑中引入一个可再现的中风模型，具有最小的侵入性。本文提供了有关手术技术的信息以及微型猪中灰质和白质的缺血性损害的方法。第二个目的是将这种新的中风模型应用于神经元再生的转化研究中，通过将teStem细胞trasplantation用于缺血性脑。方法将16个男性微型猪随机分为三组，并接受了经颅手术，这是一种递延的方法，具有轨道边缘截骨术的额叶方法，用于：永久性大脑中部动脉闭塞（MCAO）（MCAO）（N = 5），永久性内部颈内动脉（icao）（ICAO）（ICAO）（ICAO）（iCao）（n = 6）和Shams（N = 5）（n = 5）。组织学映射和磁共振（MR）成像用于描绘缺血区域。在碱性成纤维细胞生长因子存在8天的情况下，在ISCOVE修饰的Dulbecco的培养基中培养了从迷你猪中分离出来的脑室区（SVZ）细胞。我们定位移植的脑室下区（SVZ）细胞由铁磁颗粒标记为抚摸的微型猪。使用磁共振成像（MRI）对移植细胞进行非侵入性跟踪。为了检测宿主大脑中的铁磁标记的SVZ细胞，我们使用普鲁士蓝色反应染色了铁或冠状切片。结果在MCAO，ICAO和假手术组中，从MR图像直接测得的梗塞的体积分别为16.2±1.1、1.5±0.5和0.0±0.0 cm^3（平均值±SD）。组织学和MR成像量显示出良好的相关性（R^2 = 0.86，p <0.0001），尽管通过定量组织病理学测量的梗塞体积小于MRI测量的梗死体积。使用淀粉样蛋白前体蛋白抗体（APP）进行免疫组织化学染色，使我们能够评估轴突损伤，从而在MCAO中提供了43.8±3.5（平均±SD）的总APP评分。 MRI信号在病变处检测到具有标记的tranplant细胞，并且在移植后1和2周检测到这些信号。在移植细胞的同一病变中，还检测到普鲁士蓝阳性细胞。移植的细胞可能只是在宿主大脑中归巢。我们既没有证实移植细胞和宿主大脑之间网络的构建。结论本文介绍了一种简单且可重复的方法，用于诱导局灶性脑缺血中白质和灰质损害，并具有最小的侵入性。这种微型猪中风模型具有研究脑脑大脑缺血的病理生理学和评估药物或干细胞移植的治疗疗效，在开始人类临床试验之前。较少的

项目成果

期刊论文数量（18）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

New Model of Focal Cerebral Ischemia in the Miniature Pig.

小型猪局灶性脑缺血的新模型。

DOI：
发表时间：
2006
期刊：
J Neurosurg (2 Suppl Pediatrics) 104
影响因子：
0
作者：
H Imai;K Konno;M Nakamura;T Shimizu;C Kubota;K Seki;F Honda;S Tomizawa;Y Tanaka;H Hata;N Saito.
通讯作者：
N Saito.

Evolution of GADD34 expression after focal cerebral ischaemia.

局灶性脑缺血后 GADD34 表达的演变。

DOI：
发表时间：
2005
期刊：
Brain Res 1034
影响因子：
0
作者：
McCaig D;Imai H;Gallagher L;Graham DI;Harland J;Moira Brown S;Mhairi Macrae I
通讯作者：
Mhairi Macrae I

Cilostazol Attenuates Both Gray and White Matter Damage in a Rodent Model of Focal Cerebral Ischemia.

西洛他唑可减轻局灶性脑缺血啮齿动物模型中的灰质和白质损伤。

DOI：
发表时间：
2006
期刊：
Stroke 37
影响因子：
0
作者：
F Honda;H Imai;M Ishikawa;CKubota;T Shimizu;M Fukunaga;N Saito.
通讯作者：
N Saito.

The detection a d quantification of highly reactive oxygen species using the novel HPF fluorescence probe in a rat model of focal cerebral ischemia.

使用新型 HPF 荧光探针在局灶性脑缺血大鼠模型中检测和定量高活性氧。

DOI：
发表时间：
2005
期刊：
Neuroscience Research 53
影响因子：
0
作者：
S Tomizawa;H Imai;S Tsukada;T Shimizu;F Honda;M Nakamura;T Nagano;Y Ureno;Y Matsuoka;N Fukasaku;N Saito.
通讯作者：
N Saito.

Focal Cerebral ischemia increases type2 iodothyronine deiodinase.

局灶性脑缺血会增加 2 型碘甲状腺原氨酸脱碘酶。

DOI：
发表时间：
2005
期刊：
J Cereb Blood Flow Metab. 25
影响因子：
0
作者：
K Seki;H Imai;T Shimizu;K Tsunekawa;T Kasahara;T Ogiwara;N Saito;M Murakami.
通讯作者：
M Murakami.