Laboratory for Histology, Faculty of Medicine, Ghent University, L. Pasteurlaan 2, B-9000, Gent, Belgium.
Cytotechnology. 2000 Nov;34(3):185-95. doi: 10.1023/A:1008168930905.
An organotypic in vitro model, to study vascular tissueremodeling, was evaluated as a function of culture period. Inorder to validate the model as a tool for studying vascularresponses to damage, a dose-response analysis to ionizingirradiation was included.Rat aortic rings were explanted in vitro after being irradiatedwith single doses of (60)Co gamma-rays, namely 0, 5, 10, 15, 20or 25 Gy. Irradiated and sham-irradiated aortic rings werecultured for 3 weeks. Explant outgrowth on an adhesivesubstrate was evaluated by macroscopical scoring, and ringsderived from each irradiation group together with theoutgrowths were fixed and embedded in paraffin after 2, 7, 14and 21 days. Bromodeoxyuridine incorporation, alpha smoothmuscle actin and collagen types I and III were scored onimmunohistochemically stained sections. For each studiedparameter, irradiated and sham-irradiated rings were compared.In cultures of sham-irradiated rings, alterations from acontractile towards a synthetic/migratory smooth muscle cellphenotype were confirmed. After 3 weeks, fullgrown cultures hadformed. Irradiation slowed down the phenotypical modifications.After 15 Gy, irradiation explant outgrowth was already retarded;after 25 Gy, the outgrowth was completely blocked. On the otherhand, a dose of 15 Gy or more induced an increased collagen Iproduction in the tunica media.In conclusion, the present organotypical in vitro model fits toanalyse dynamics in the original vascular tissues as well as inthe primary outgrowth. It enables to confirm features oftissular reorganization and effects of ionizing radiationdescribed in vivo.
建立了一种器官型体外模型，用于研究血管组织重构，该模型可以作为评估的功能，随培养时间的变化而变化。为了验证该模型作为研究血管对损伤反应的工具，包括了对电离辐射的剂量反应分析。在接受单次(60)Coγ射线照射后，将大鼠主动脉环在体外进行离体培养，剂量分别为 0、5、10、15、20 或 25Gy。照射和假照射的主动脉环培养 3 周。通过肉眼评分评估贴壁培养的外生体生长情况，并且在 2、7、14 和 21 天之后，将来自每个照射组的环和外生体固定并嵌入石蜡中。溴脱氧尿苷掺入、α平滑肌肌动蛋白和胶原 I 型和 III 型在免疫组织化学染色切片上进行评分。对于每个研究的参数，比较了照射和假照射的环。在假照射环的培养中，从收缩性向合成/迁移平滑肌细胞表型的改变得到了证实。3 周后，完全生长的培养物形成。照射减缓了表型的改变。15Gy 后，照射外生体生长已经受到抑制；25Gy 后，外生体完全被阻断。另一方面，15Gy 或更高的剂量会导致中膜胶原 I 的产生增加。总之，这种器官型体外模型适合分析原始血管组织以及原发性外生体的动力学。它可以证实体内描述的组织重构特征和电离辐射的作用。
