Deborde Christopher, Simionescu Dan Teodor, Wright Cristopher, Liao Jun, Sierad Leslie Neil, Simionescu Agneta
1 Department of Bioengineering, Clemson University , Clemson, South Carolina.
2 Department of Cardiothoracic Surgery, Greenville Memorial Hospital , Greenville, South Carolina.
Tissue Eng Part A. 2016 Nov;22(21-22):1241-1251. doi: 10.1089/ten.TEA.2016.0032. Epub 2016 Oct 3.
There is a significant clinical need for new approaches to treatment of mitral valve disease. The aim of this study was to develop a tissue-engineered mitral valve scaffold possessing appropriate composition and structure to ensure ideal characteristics of mitral valves, such as large orifice, rapid opening and closure, maintenance of mitral annulus-papillary muscle continuity, in vivo biocompatibility and extended durability. An extracellular matrix-based scaffold was generated, based on the native porcine mitral valve as starting material and a technique for porcine cell removal without causing damage to the matrix components. To stabilize these structures and slow down their degradation, acellular scaffolds were treated with penta-galloyl glucose (PGG), a well-characterized polyphenol with high affinity for collagen and elastin. Biaxial mechanical testing presented similar characteristics for the PGG-treated scaffolds compared to fresh tissues. The extracellular matrix components, crucial for maintaining the valve shape and function, were well preserved in leaflets, and in chordae, as shown by their resistance to collagenase and elastin. When extracted with strong detergents, the PGG-treated scaffolds released a reduced amount of soluble matrix peptides, compared to untreated scaffolds; this correlated with diminished activation of fibroblasts seeded on scaffolds treated with PGG. Cell-seeded scaffolds conditioned for 5 weeks in a valve bioreactor showed good cell viability. Finally, rat subdermal implantation studies showed that PGG-treated mitral valve scaffolds were biocompatible, nonimmunogenic, noninflammatory, and noncalcifying. In conclusion, a biocompatible mitral valve scaffold was developed, which preserved the biochemical composition and structural integrity of the valve, essential for its highly dynamic mechanical demands, and its biologic durability.
二尖瓣疾病的治疗急需新方法。本研究的目的是开发一种组织工程二尖瓣支架,其具有合适的组成和结构,以确保二尖瓣具备理想特性,如大开口、快速开闭、维持二尖瓣环-乳头肌连续性、体内生物相容性和延长的耐久性。基于天然猪二尖瓣作为起始材料,并采用一种在不损伤基质成分的情况下去除猪细胞的技术,生成了一种基于细胞外基质的支架。为了稳定这些结构并减缓其降解,对脱细胞支架用五倍子酰葡萄糖(PGG)进行处理,PGG是一种对胶原蛋白和弹性蛋白具有高亲和力且特性明确的多酚。与新鲜组织相比,双轴力学测试显示PGG处理的支架具有相似的特性。对维持瓣膜形状和功能至关重要的细胞外基质成分在瓣叶和腱索中保存良好,这通过它们对胶原酶和弹性蛋白酶的抗性得以体现。与未处理的支架相比,用强去污剂提取时,PGG处理的支架释放的可溶性基质肽量减少;这与接种在PGG处理支架上的成纤维细胞活化减少相关。在瓣膜生物反应器中培养5周的接种细胞支架显示出良好的细胞活力。最后,大鼠皮下植入研究表明,PGG处理的二尖瓣支架具有生物相容性、无免疫原性、无炎症且无钙化。总之,开发了一种生物相容性二尖瓣支架,其保留了瓣膜的生化组成和结构完整性,这对于其高度动态的力学需求及其生物学耐久性至关重要。