Department of Bioengineering, University of Pittsburgh, Pittsburgh, PA 15261, USA.
Proc Natl Acad Sci U S A. 2011 Feb 15;108(7):2705-10. doi: 10.1073/pnas.1017834108. Epub 2011 Jan 31.
Mature elastin synthesis is a key challenge in arterial tissue engineering. Most engineered vessels lack elastic fibers in the medial layer and those present are poorly organized. The objective of this study is to increase mature elastin synthesis in small-diameter arterial constructs. Adult primary baboon smooth muscle cells (SMCs) were seeded in the lumen of porous tubular scaffolds fabricated from a biodegradable elastomer, poly(glycerol sebacate) (PGS) and cultured in a pulsatile flow bioreactor for 3 wk. We tested the effect of pore sizes on construct properties by histological, biochemical, and mechanical evaluations. Histological analysis revealed circumferentially organized extracellular matrix proteins including elastin and the presence of multilayered SMCs expressing calponin and α-smooth muscle actin. Biochemical analysis demonstrated that the constructs contained mature elastin equivalent to 19% of the native arteries. Mechanical tests indicated that the constructs could withstand up to 200 mmHg burst pressure and exhibited compliance comparable to native arteries. These results show that nontransfected cells in PGS scaffolds in unsupplemented medium produced a substantial amount of mature elastin within 3 wk and the elastic fibers had similar orientation as those in native arteries. The 25-32 μm pore size supported cell organization and elastin synthesis more than larger pore sizes. To our knowledge, there was no prior report of the synthesis of mature and organized elastin in arterial constructs without exogenous factors or viral transduction.
成熟弹性蛋白的合成是动脉组织工程的一个关键挑战。大多数工程化血管缺乏中膜层的弹性纤维,而存在的纤维组织排列不良。本研究的目的是增加小直径动脉构建体中成熟弹性蛋白的合成。将成年原代狒狒平滑肌细胞(SMCs)接种在由可生物降解弹性体聚(甘油-癸二酸酯)(PGS)制成的多孔管状支架的内腔中,并在脉动流生物反应器中培养 3 周。我们通过组织学、生物化学和力学评估来测试孔径对构建体性能的影响。组织学分析显示,圆周排列的细胞外基质蛋白包括弹性蛋白,以及表达钙调蛋白和平滑肌肌动蛋白-α的多层 SMC。生物化学分析表明,构建体含有相当于 19%的天然动脉的成熟弹性蛋白。力学测试表明,构建体可以承受高达 200mmHg 的爆裂压力,并表现出与天然动脉相当的顺应性。这些结果表明,在未补充培养基的 PGS 支架中,未经转染的细胞在 3 周内产生了大量的成熟弹性蛋白,并且弹性纤维的取向与天然动脉中的相似。25-32μm 的孔径比较大的孔径更支持细胞组织和弹性蛋白的合成。据我们所知,在没有外源因子或病毒转导的情况下,以前没有关于在动脉构建体中合成成熟和有组织的弹性蛋白的报道。