Biomedical Engineering Program, College of Engineering, University of South Carolina, Columbia, South Carolina, USA.
Department of Cell Biology and Anatomy, School of Medicine, University of South Carolina, Columbia, South Carolina, USA.
J Biomed Mater Res B Appl Biomater. 2022 Apr;110(4):885-897. doi: 10.1002/jbm.b.34969. Epub 2021 Dec 2.
Engineered replacement materials have tremendous potential for vascular applications where over 400,000 damaged and diseased blood vessels are replaced annually in the United States alone. Unlike large diameter blood vessels, which are effectively replaced by synthetic materials, prosthetic small-diameter vessels are prone to early failure, restenosis, and reintervention surgery. We investigated the differential response of varying 0%-6% sodium dodecyl sulfate and sodium deoxycholate anionic detergent concentrations after 24 and 72 h in the presence of DNase using biochemical, histological, and biaxial mechanical analyses to optimize the decellularization process for xenogeneic vascular tissue sources, specifically the porcine internal thoracic artery (ITA). Detergent concentrations greater than 1% were successful at removing cytoplasmic and cell surface proteins but not DNA content after 24 h. A progressive increase in porosity and decrease in glycosaminoglycan (GAG) content was observed with detergent concentration. Augmented porosity was likely due to the removal of both cells and GAGs and could influence recellularization strategies. The treatment duration on the other hand, significantly improved decellularization by reducing DNA content to trace amounts after 72 h. Prolonged treatment times reduced laminin content and influenced the vessel's mechanical behavior in terms of altered circumferential stress and stretch while further increasing porosity. Collectively, DNase with 1% detergent for 72 h provided an effective and efficient decellularization strategy to be employed in the preparation of porcine ITAs as bypass graft scaffolding materials with minor biomechanical and histological penalties.
工程替代材料在血管应用方面具有巨大的潜力,仅在美国,每年就有超过 40 万个受损和患病的血管需要更换。与大直径血管不同,大直径血管可以有效地用合成材料替代,而人造小直径血管容易早期失效、再狭窄和再次介入手术。我们研究了不同浓度(0%-6%)十二烷基硫酸钠和脱氧胆酸钠阴离子去污剂在存在 DNA 酶的情况下,经过 24 小时和 72 小时后对小直径血管的影响,采用生物化学、组织学和双向力学分析来优化异种血管组织来源(特别是猪的胸内动脉)的脱细胞过程。浓度大于 1%的去污剂在 24 小时后成功去除细胞质和细胞膜蛋白,但不能去除 DNA 含量。随着去污剂浓度的增加,孔隙率逐渐增加,糖胺聚糖(GAG)含量逐渐减少。孔隙率的增加可能是由于细胞和 GAG 的去除,这可能会影响再细胞化策略。另一方面,处理时间延长 72 小时后,通过将 DNA 含量降低到痕量,显著提高了脱细胞效率。延长处理时间会降低层粘连蛋白含量,并影响血管的力学性能,表现为环向应力和拉伸的改变,同时进一步增加孔隙率。总的来说,用 1%去污剂和 DNA 酶处理 72 小时是一种有效且高效的脱细胞策略,可以用于制备猪胸内动脉作为旁路移植物支架材料,对生物力学和组织学影响较小。