J. Crayton Pruitt Family Department of Biomedical Engineering, University of Florida, Gainesville, Florida 32611, USA.
J Biomed Mater Res A. 2013 Jan;101(1):123-31. doi: 10.1002/jbm.a.34308. Epub 2012 Jul 24.
The use of ex vivo-derived scaffolds as vascular conduits has shown to be a clinically valid approach to repair or bypass occluded vessels. Implantation of allogeneic tissue grafts requires careful processing to lower immunogenicity and prevent bacterial infection. However, the mechanical/chemical treatments used to prepare biological scaffolds can result in significant alterations to the native structure and surface chemistry, which can affect in vivo performance. Of particular importance for vascular grafts are binding interactions between the implanted biomaterial and host cells from the circulation and adjacent vasculature. Here we present a comparison of four strategies used to decellularize allogeneic human umbilical vein (HUV) scaffolds: ethanol/acetone, sodium chloride, sodium dodecyl sulfate (SDS), or Triton X-100. Scanning electron microscopy revealed that all four techniques achieved removal of native cells from both the lumenal and ablumenal surfaces of HUV grafts. Platelets and promyelocytic HL-60 cells showed preferential binding on the more loosely structured ablumenal surface, although low surface coverage was observed overall by peripheral blood cells. Vascular endothelial cell adhesion was highest on HUV decellularized using ethanol/acetone, and significantly higher than on SDS-processed grafts (p = 0.016). Primary cells showed high viability on the lumenal surface regardless of decellularization technique (over 95% in all cases). These results demonstrate the critical effects of various chemical processing strategies on the adhesive properties of ex vivo-derived vascular grafts. Careful application-specific consideration is warranted when selecting a processing strategy that minimizes innate responses (e.g. thrombosis, inflammation) that are often deleterious to graft survival.
将从体外分离得到的支架作为血管移植物已被证明是修复或旁路阻塞血管的一种有效的临床方法。同种异体组织移植物的植入需要仔细处理以降低免疫原性并防止细菌感染。然而,用于制备生物支架的机械/化学处理可能会导致对天然结构和表面化学性质的显著改变,从而影响体内性能。对于血管移植物来说,特别重要的是植入生物材料与来自循环系统和相邻血管的宿主细胞之间的结合相互作用。在这里,我们比较了四种用于脱细胞化同种异体人脐静脉(HUV)支架的策略:乙醇/丙酮、氯化钠、十二烷基硫酸钠(SDS)或 Triton X-100。扫描电子显微镜显示,这四种技术都能从 HUV 移植物的腔面和腔外表面去除天然细胞。血小板和早幼粒细胞 HL-60 细胞优先在结构较松散的腔外表面上结合,但外周血细胞的总结合面积较低。血管内皮细胞在使用乙醇/丙酮脱细胞化的 HUV 上的粘附性最高,显著高于 SDS 处理的移植物(p = 0.016)。无论脱细胞化技术如何,内皮细胞在腔面都具有很高的活力(所有情况下均超过 95%)。这些结果表明,各种化学处理策略对体外衍生血管移植物的粘附特性具有重要影响。在选择一种处理策略时,应根据具体情况进行仔细考虑,以尽量减少对移植物存活有害的固有反应(如血栓形成、炎症)。