کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
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6269247 | 1295129 | 2012 | 5 صفحه PDF | دانلود رایگان |

The blood-brain barrier (BBB) restricts the delivery of drugs into the brain. Different strategies have been developed to circumvent this obstacle. One such approach, the osmotic BBB disruption (BBBD), has been under pre-clinical study since the 70's. Typically, qualitative ex vivo assessment of the extent of BBBD has been performed using Evan's blue staining technique. In this study, we describe a simple quantitative technique based on albumin indirect immunohistochemistry to measure the extent of BBB breach. Thirty Fischer rats were assigned to one of 6 groups: a control group, and BBBD groups with escalation in IA mannitol infusion rate: 0.06, 0.08, 0.10, 0.12 and 0.15Â cc/s. Fifteen minutes after the BBBD procedure, the animals were sacrificed, brain harvested and sections stained for albumin. Using an image analysis software, isolated albumin staining pixels were expressed as a fraction of the treated hemisphere. This ratio was used as a percentage value in the intensity of the BBB permeabilization. All sections studied harbored staining, averaging 0.37% for the controls (group 1), 5.69% for group 2 (0.06Â cc/s), 10.44% for group 3 (0.08Â cc/s), 6.99% for group 4 (0.1Â cc/s), 18.50% for group 5 (0.12Â cc/s) and reaching 61.70% for group 6 (0.15Â cc/s). Important variations were observed between animals. A threshold effect was observed, and animals in group 6 presented a significant increase in BBB permeabilization compared to the other groups. We hereby detail a simple technique that can be applied to quantitatively measure the extent of the BBB breach notwithstanding the pathological process.
⺠We describe a new quantitative method to assess the extent of the BBB disruption. ⺠The method relies on the detection of albumin by indirect-immunohistochemistry. ⺠The extent of the BBBD was studied using increasing mannitol infusion rates. ⺠At 0.15 cc/s, we observed a threshold effect on the BBB permeability.
Journal: Journal of Neuroscience Methods - Volume 207, Issue 2, 15 June 2012, Pages 125-129