Nguyen Thuy-Uyen, Bashur Chris A, Kishore Vipuil
Department of Chemical Engineering, Florida Institute of Technology, Melbourne, FL 32901, USA.
Biomed Mater. 2016 Mar 17;11(2):025008. doi: 10.1088/1748-6041/11/2/025008.
Application of tissue-engineered vascular grafts (TEVGs) for the replacement of small-diameter arteries is limited due to thrombosis and intimal hyperplasia. Previous studies have attempted to address the limitations of TEVGs by developing scaffolds that mimic the composition (collagen and elastin) of native arteries to better match the mechanical properties of the graft with the native tissue. However, most existing scaffolds do not recapitulate the aligned topography of the collagen fibers found in native vessels. In the current study, based on the principles of isoelectric focusing, two different types of elastin (soluble and insoluble) were incorporated into highly oriented electrochemically aligned collagen (ELAC) fibers and the effect of elastin incorporation on the mechanical properties of the ELAC fibers and smooth muscle cell (SMC) phenotype was investigated. The results indicate that elastin incorporation significantly decreased the modulus of ELAC fibers to converge upon that of native vessels. Further, a significant increase in yield strain and decrease in Young's modulus was observed on all fibers post SMC culture compared with before the culture. Real-time polymerase chain reaction results showed a significant increase in the expression of α-smooth muscle actin and calponin on ELAC fibers with insoluble elastin, suggesting that incorporation of insoluble elastin induces a contractile phenotype in SMCs after two weeks of culture on ELAC fibers. Immunofluorescence results showed that calponin expression increased with time on all fibers. In conclusion, insoluble elastin incorporated ELAC fibers have the potential to be used for the development of functional TEVGs for the repair and replacement of small-diameter arteries.
由于血栓形成和内膜增生,组织工程血管移植物(TEVG)在小直径动脉置换中的应用受到限制。先前的研究试图通过开发模仿天然动脉成分(胶原蛋白和弹性蛋白)的支架来解决TEVG的局限性,以使移植物的机械性能更好地与天然组织相匹配。然而,大多数现有的支架并未重现天然血管中发现的胶原纤维的排列形貌。在本研究中,基于等电聚焦原理,将两种不同类型的弹性蛋白(可溶性和不溶性)掺入高度取向的电化学排列胶原(ELAC)纤维中,并研究了弹性蛋白掺入对ELAC纤维机械性能和平滑肌细胞(SMC)表型的影响。结果表明,弹性蛋白的掺入显著降低了ELAC纤维的模量,使其与天然血管的模量趋于一致。此外,与培养前相比,在SMC培养后所有纤维上均观察到屈服应变显著增加,杨氏模量降低。实时聚合酶链反应结果显示,含有不溶性弹性蛋白的ELAC纤维上α-平滑肌肌动蛋白和钙调蛋白的表达显著增加,表明在ELAC纤维上培养两周后,不溶性弹性蛋白的掺入诱导了SMC中的收缩表型。免疫荧光结果显示,所有纤维上钙调蛋白的表达均随时间增加。总之,含有不溶性弹性蛋白的ELAC纤维有潜力用于开发功能性TEVG,以修复和置换小直径动脉。