利用低持续腔内压力将人脂肪来源的间质血管部分细胞转移到ePTFE上制备组织工程血管移植物

H. J. Paek, Stuart K. Williams, A. Yang, Courtney Kim, Shannon Iwami, Todd Case, S. Berman, Eugene Bol, P. Kosnik
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摘要

每年有数百万美国人受到心血管和外周动脉疾病的影响。当患者自身的血管不适合手术干预时,就需要替代的移植物来源。在本研究中,利用持续低腔内压力,用新分离的脂肪源性基质血管组分(SVF)细胞或培养的脂肪源性基质细胞(ASCs)包裹膨化聚四氟乙烯(ePTFE)导管的管腔。ePTFE血管导管在短短5分钟内被铺设。通过对导管上附着细胞的核染色来评估压力灌注和随后暴露于生理管腔流后移植物的管腔表面覆盖率。通过组织因子量化,研究了细胞行为与灌注过程和随后的腔流剪切应力的关系。在以近似人类冠状动脉的生理剪切速率暴露于腔内血流后,被灌注的细胞仍留在ePTFE导管上。当使用培养的SVF细胞(ASCs,附着的基质细胞)时,平均标准化组织因子水平随着细胞灌注过程后管腔流的应用而增加。然而,当使用新鲜分离的SVF细胞时,即使经过管腔流动,平均归一化组织因子也没有显着变化。当这些细胞包裹的血管移植物被植入狗的颈动脉时,它们至少保持140天的专利。这种新颖的压力灌注方法对于快速制造组织工程血管移植物至关重要,它可以促进许多血管疾病的成功护理点治疗。
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Fabrication of tissue-engineered vascular grafts using low sustained intraluminal pressure to sod human adipose-derived stromal vascular fraction cells onto ePTFE
Every year millions of Americans are affected by cardiovascular and peripheral arterial diseases. When a patient’s own blood vessels are not suitable for surgical intervention, alternative graft sources are necessary. In this study, the lumens of expanded polytetrafluoroethylene (ePTFE) conduits were coated with freshly isolated adipose-derived stromal vascular fraction (SVF) cells or cultured adipose-derived stromal cells (ASCs) by a single-stage method called “pressure sodding,” using low sustained intraluminal pressure. ePTFE vascular conduits were sodded in as short as 5 minutes. The luminal surface coverage of grafts immediately following pressure sodding and subsequent exposure to physiological luminal flow was evaluated by nuclear staining of the attached cells to the conduit. Cell behavior associated with the sodding process and subsequent shear stress via luminal flow was examined through tissue factor quantification. After exposure to luminal flow at a rate approximating physiological shear in human coronary arteries, the sodded cells remained on the ePTFE conduits. When cultured SVF cells (ASCs, attached stromal cells) were used, average normalized tissue factor levels increased with the application of a luminal flow following the cell sodding process. However, when freshly isolated SVF cells were used, the average normalized tissue factor did not significantly change even after luminal flow. When these cell-coated vascular grafts were implanted into the carotid arteries of dogs, they remained patent for at least 140 days. This novel method of pressure sodding is critical to rapid fabrication of tissue-engineered vascular grafts which can facilitate a successful point-of-care treatment for a number of vascular diseases.
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