Ross Claire, Byard Roger W, Langlois Neil E I
School of Medical Sciences, The University of Adelaide, Forensic Science SA, 21 Divett Place, Adelaide, SA, Australia.
Forensic Sci Med Pathol. 2013 Sep;9(3):358-62. doi: 10.1007/s12024-013-9466-y. Epub 2013 Jul 10.
Whole blood was withdrawn by tail vessel puncture from anesthetized adult male Sprague-Dawley rats and 0.1 ml was re-injected subcutaneously at each of two sites on their abdominal wall. In addition, two adjacent sites were injected with 0.1 ml of sterile saline, and two more sites were only punctured using an injecting needle. In the second part of the study anesthetized adult male Sprague-Dawley rats had two sites on the abdominal wall pinched using a small pair of forceps, two adjacent sites received an injection of 0.1 ml of whole blood obtained by tail vessel puncture, and two more sites were both pinched and injected with 0.1 ml of whole blood. At intervals of 3, 6, 12 h, 1, 2, 3, 5, and 7 days the animals were euthanized and the skin of the abdomen was processed for histological assessment. Hemosiderin staining in tissues from the first part of the study was assessed qualitatively by scoring sections as 0, 1, 2, or 3 (representing no staining, mild staining, moderate staining, and intense staining) and semi quantitatively using a Nanozoomer Digital Pathology Scanner (NDP Scan U10074-01, Hamamatsu Photonics K.K., Japan). No inflammatory reaction was observed at the sites subjected to needle puncture only. At the sites of saline injection a mild reaction occurred. At the sites where the blood had been injected an intense inflammatory cell response occurred centrally, but not toward the periphery where blood had tracked. In the second experiment the most intense inflammation was also observed in the sites where there had been a pinch and injection of blood. Again, this was maximal centrally with reduced inflammation peripherally. Perls' staining of hemosiderin was comparable in both models, with iron first observed at day 1 at the region of the injection site. At the sites of injection only, and the sites of injection plus pinch, blood had spread laterally. Hemosiderin staining appeared first and more intensely at the site of injection/trauma. The intensity of the inflammatory response in this animal model of bruising was, therefore, directly related to the proximity to the site of trauma; the appearance and intensity of hemosiderin staining was also influenced by the location within the bruises. This study has shown that histological changes that may be utilized to date bruises may be significantly influenced by the site of the biopsy.
通过尾静脉穿刺从麻醉的成年雄性Sprague-Dawley大鼠采集全血，并将0.1 ml全血分别皮下注射到其腹壁的两个部位。此外，在两个相邻部位注射0.1 ml无菌生理盐水，另外两个部位仅用注射针刺破。在研究的第二部分，用一把小镊子夹住麻醉的成年雄性Sprague-Dawley大鼠腹壁的两个部位，两个相邻部位注射0.1 ml通过尾静脉穿刺获得的全血，另外两个部位既被夹住又注射0.1 ml全血。在3、6、12小时、1、2、3、5和7天的时间间隔，对动物实施安乐死，并对腹部皮肤进行组织学评估。研究第一部分组织中的含铁血黄素染色通过将切片评为0、1、2或3（分别代表无染色、轻度染色、中度染色和重度染色）进行定性评估，并使用Nanozoomer数字病理扫描仪（NDP Scan U10074-01，日本浜松光子株式会社）进行半定量评估。仅接受针刺的部位未观察到炎症反应。在注射生理盐水的部位出现了轻度反应。在注射血液的部位，中央出现强烈的炎症细胞反应，但在血液蔓延的周边部位未出现。在第二个实验中，在既被夹住又注射血液的部位也观察到最强烈的炎症。同样，炎症在中央最为严重，周边炎症减轻。两种模型中含铁血黄素的Perls染色相当，在注射部位区域第1天首次观察到铁。仅在注射部位以及注射加夹住的部位，血液发生了侧向扩散。含铁血黄素染色首先在注射/创伤部位出现且更为强烈。因此，在这个瘀伤动物模型中，炎症反应的强度与距创伤部位的距离直接相关；含铁血黄素染色的出现和强度也受瘀伤内位置的影响。本研究表明，可用于确定瘀伤时间的组织学变化可能会受到活检部位的显著影响。
