Hodonsky Chani, Mundada Lakshmi, Wang Shuyun, Witt Russell, Raff Gary, Kaushal Sunjay, Si Ming-Sing
Department of Cardiac Surgery, University of Michigan, Ann Arbor, Michigan.
Department of Surgery, University of California at Davis Medical Center, Sacramento, California.
Ann Thorac Surg. 2015 Feb;99(2):605-11. doi: 10.1016/j.athoracsur.2014.08.071. Epub 2014 Dec 12.
Polytetrafluoroethylene (PTFE) and porcine small intestinal submucosa (pSIS) are patch materials used in congenital heart surgery. Porcine SIS is an extracellular-matrix scaffold that may interact with stem or progenitor cells. To evaluate this, we determined the in vitro effects of pSIS and PTFE on human bone marrow mesenchymal stromal cells (MSCs) and cardiac progenitor cells (CPCs) in 3 areas; cell proliferation, angiogenic growth-factor production, and differentiation.
Human MSCs and CPCs were seeded onto pSIS and PTFE patches. Cell-seeded patches were cultured and then assessed for cell viability and proliferation and supernatant vascular endothelial growth factor A (VEGFA) levels. Cell proliferation was quantified by MTT assay (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide). Quantitative real-time polymerase chain reaction was performed on cell-seeded scaffolds to determine relative changes in gene expression related to angiogenesis and cardiogenesis.
The MSCs and CPCs were able to attach and proliferate on pSIS and PTFE. The proliferation rate of each cell type was similar on pSIS. Total RNA isolation was only possible from the cell-seeded pSIS patches. The MSC VEGFA production was increased by pSIS. Porcine SIS promoted an angiogenic gene profile in MSCs and an early cardiogenic profile in CPCs.
Both PTFE and pSIS allow for varying degrees of cell proliferation. Porcine SIS elicits different phenotypical responses in MSCs as compared with CPCs, which indicates that pSIS may be a bioactive scaffold that modulates stem cell activation and proliferation. These findings highlight the differences in scaffold material strategies and suggest potential advantages of bioactive approaches.
聚四氟乙烯(PTFE)和猪小肠黏膜下层(pSIS)是用于先天性心脏病手术的补片材料。猪SIS是一种细胞外基质支架,可能与干细胞或祖细胞相互作用。为了评估这一点,我们在三个方面确定了pSIS和PTFE对人骨髓间充质基质细胞(MSCs)和心脏祖细胞(CPCs)的体外影响;细胞增殖、血管生成生长因子的产生和分化。
将人MSCs和CPCs接种到pSIS和PTFE补片上。接种细胞的补片进行培养,然后评估细胞活力、增殖情况以及上清液中血管内皮生长因子A(VEGFA)水平。通过MTT法(3-[4,5-二甲基噻唑-2-基]-2,5-二苯基四氮唑溴盐)对细胞增殖进行定量。对接种细胞的支架进行定量实时聚合酶链反应,以确定与血管生成和心脏发生相关的基因表达的相对变化。
MSCs和CPCs能够在pSIS和PTFE上附着并增殖。每种细胞类型在pSIS上的增殖率相似。仅从接种细胞的pSIS补片上能够分离出总RNA。pSIS可增加MSC的VEGFA产生。猪SIS促进MSCs中的血管生成基因谱和CPCs中的早期心脏发生基因谱。
PTFE和pSIS都允许不同程度的细胞增殖。与CPCs相比,猪SIS在MSCs中引发不同的表型反应,这表明pSIS可能是一种调节干细胞活化和增殖的生物活性支架。这些发现突出了支架材料策略的差异,并提示了生物活性方法的潜在优势。