染料介导光氧化固定牛颈静脉血管片体外内皮化

目的研究染料介导光氧化固定牛颈静脉的细胞毒性．初步探讨此种材料体外内皮化的可能性，方法实验组为染料介导光氧化固定组。对照组为戊二醛固定组。采用羊的颈内静脉内皮细胞作为细胞源，细胞种植于管腔一侧，时间为5d。动态观察细胞生长情况，扫描电子显微镜检查种植细胞前后的材料和免疫组织化学因子Ⅷ染色情况。结果光氧化固定组．采用扫描电子显微镜观察，在种植细胞前，在材料表面无细胞，而在种植细胞后，在材料表面密布一层细胞。戊二醛固定组．采用扫描电子显微镜观察，在种植细胞前后无差别，在材料表面见戊二醛固定时残留的少许细胞。光氧化同定组．材料表面种植的细胞因子Ⅷ染色为阳性。结论染料介导光氧化固定牛颈静脉血管片无明显细胞毒性．在体外可以很好地内皮化。

In vitro endothelialization of photooxidatively stabilized bovine jugular veins

Objective To evaluate the cytotoxicity of dye-mediated photooxidatively stabilized bovine jugular vein and its possibility of endothelialization in vitro. Methods The glutaraldehyde stabilized bovine jugular veins were enrolled as control. In the photooxidation group, tissue samples were immersed in a chilled phosphate buffered solution（PBS） containing methylene blue and exposed to the incandescent light for 96 hours. After fixation, the tissue was washed throughly to remove excess dye, sterilized in an iodine solution, and stored in 50 % ethanol. In the glutaraldehyde group, the tissue samples were immersed in 0.6 % glutaraldehyde for 2 days and stored in 0.3 % glutaraldehyde. The tissue samples were seeded with cultured adult ovine jugular vein endothelial cells. Scanning electron microscopy, hematoxylin-eosin and immunohistochemical staining for the factor Ⅷ were performed in sections of endothelialized bovine jugular patches. Results In the photooxidation group, a confluent lining of cultured ovine jugular vein endothelial cells similar to natural endothelium was demonstrated by scanning electron microscopy, and the presence of factor Ⅷwas demonstrated by using monoclonal antibodies. In the glutaraldehyde group, there were no cultured ovine jugular vein endothelial cells growing on the tissue samples under scanning electron microscopic observation. Conclusion The results clearly indicate that the dye-mediated photooxidation technique produces a tissue without cytotoxicity, and the in vitro endothelialization of such tissue may be realized.