Rupnick M A, Hubbard F A, Pratt K, Jarrell B E, Williams S K
Department of Physiology, Jefferson Medical College, Thomas Jefferson University, Philadelphia, PA 19107.
J Vasc Surg. 1989 Jun;9(6):788-95. doi: 10.1067/mva.1989.vs0090788.
The rapid establishment of an endothelial cell (EC) monolayer on the luminal surface of small-diameter vascular grafts may be necessary to prevent early thrombosis and failure. We have studied procedures used to promote EC coverage of vascular grafts and have compared preclotting prosthetic surfaces with ECs in platelet-rich plasma (seeding) with plating ECs onto a preestablished clot (sodding). We evaluated the rate of monolayer formation, the subsequent resistance to shear stress, and the effects of EC growth factors (ECGF and heparin) on these functions. Woven Dacron was seeded or sodded at a density of 2 x 10(5) cells/cm2 with human adult microvessel ECs derived from adipose tissue. In the presence of ECGF-heparin, the immediate establishment of an EC layer after sodding was observed, whereas seeded grafts required almost 48 hours for cells to reach the surface. In the absence of ECGF-heparin, sodded grafts still exhibited a complete monolayer of EC, whereas ECs were not observed at the surface of seeded grafts after 48 hours. After exposure to shear stress (up to 20 dynes/cm2) for 2 hours, most freshly sodded EC remained attached; however, the loss of loosely adherent cells did occur. EC seeded grafts remained covered with fibrin matrix after exposure to shear stress. We conclude that the use of a microvessel sodding technique as an alternative to previously reported seeding techniques is necessary for the immediate formation of an EC monolayer before implantation.
在小口径血管移植物的管腔表面快速形成内皮细胞(EC)单层对于预防早期血栓形成和移植物失败可能是必要的。我们研究了用于促进血管移植物内皮细胞覆盖的方法,并比较了在富含血小板血浆(接种)中用内皮细胞预凝人工血管表面与将内皮细胞铺板到预先形成的凝块上(草皮移植)的方法。我们评估了单层形成的速率、随后对剪切应力的抵抗力以及内皮细胞生长因子(内皮细胞生长因子和肝素)对这些功能的影响。将成人脂肪组织来源的人微血管内皮细胞以2×10⁵个细胞/cm²的密度接种或草皮移植到编织涤纶上。在存在内皮细胞生长因子 - 肝素的情况下,草皮移植后立即观察到内皮细胞层的形成,而接种的移植物细胞到达表面需要近48小时。在不存在内皮细胞生长因子 - 肝素的情况下,草皮移植的移植物仍表现出完整的内皮细胞单层,而接种的移植物在48小时后表面未观察到内皮细胞。在暴露于剪切应力(高达20达因/cm²)2小时后,大多数新鲜草皮移植的内皮细胞仍保持附着;然而,确实发生了松散附着细胞的丢失。接种内皮细胞的移植物在暴露于剪切应力后仍覆盖有纤维蛋白基质。我们得出结论,在植入前立即形成内皮细胞单层,使用微血管草皮移植技术作为先前报道的接种技术的替代方法是必要的。