Construction and vascularization of tissue-engineered sinus node in cardiac tissue engineering

doi:10.3969/j.issn.2095-4344.2017.16.025

Abstract

Abstract:

BACKGROUND: As in theory tissue-engineered sinus node can establish a new pacemaker in vivo when implanted into the human body, it is a promising treatment for sick sinus syndrome. But this method is immature and needs to be explored in depth.
OBJECTIVE: To overview the different construction methods of tissue-engineered sinus node, and the research progress of vascularization in cardiac tissue engineering.
METHODS: A computer-based retrieval in PubMed and CNKI databases was performed by the first author to search related papers published from 1984 to 2016 using the keywords of “vascularize, tissue engineering, sinus node” in English and Chinese, respectively. We summarized the construction methods of tissue-engineered sinus node and importance of vascularization in cardiac tissue engineering.
RESULTS AND CONCLUSION: Sinus node cells, bone marrow mesenchymal stem cells, adipose-derived mesenchymal stem cells and embryonic stem cells can be used as seed cells. There are a variety of materials used for constructing tissue-engineered scaffolds, among which, collagen is the best choice. Three-dimensional printing technology and cell-sheet techbology make it possible to construct and transplant tissue-engineered sinus node. The early blood supply is the key for the successful sinus node transplantation. However, the vascularized tissue-engineered sinus node has not yet been reported. Endothelial progenitor cells can promote angiogenesis, but further explorations are warranted as there are some existing defects.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Key words: Sinoatrial Node, Neovascularization, Physiologic, Tissue Engineering

CLC Number:

R318

Tong Ling-xi, Zhang Chuan-sen, Huang Fei. Construction and vascularization of tissue-engineered sinus node in cardiac tissue engineering[J]. Chinese Journal of Tissue Engineering Research, 2017, 21(16): 2606-2611.

Figures/Tables 1

2.1 组织工程窦房结的构建构建组织工程窦房结要力求接近生理窦房结，同时要能对血管化的形成、移植时长入机体有良好的促进作用，最重要的是让支架材料在体内降解，不对机体产生作副作用。Zimmermann等[4]在2004年提出了心脏组织工程的标准：①具备一定收缩能力；②电生理学稳定；③能够血管化；④低免疫原性。优良的组织工程窦房结也应遵守这一标准。起搏种子细胞的选择大致为窦房结原代细胞，脂肪、骨髓间充质干细胞，胚胎干细胞等。Marvin等[5]用通过对乳鼠窦房结进行定位，对窦房结细胞进行分离、纯化，培养出的细胞形态、结构均类似于窦房结细胞或心肌细胞，在电镜下观察细胞的超微结构类似于心脏的窦性节段的细胞，用膜片钳记录到该细胞的动作电位，认为其可以广泛的应用于单细胞的研究，也可以作为组织工程窦房结潜在的种子细胞。Fukuda[6]提取鼠类的骨髓间充质干细胞，将其用5-氮杂胞嘧啶核苷处理，同时根据其搏动进行筛选，在1周时细胞结构会类似于成纤维细胞，2周左右会出现自发的搏动，而在3周时搏动统一。这些细胞会同时表达脑钠肽和心钠肽在经5-氮杂胞嘧啶核苷处理前细胞表达Nkx2.5, GATA4，TEF-1， MEF2-C mRNA，而在处理后表达MEF2-A、MEF2-D的mRNA。结构上出现心肌小节和心房颗粒，具有窦房结细胞或心室肌肉细胞的潜能。张喜等[7]以11 d胎鼠为源提取胚胎心脏祖细胞，利用内皮素1诱导其为起搏细胞。免疫组织工程检测表达起搏离子通道蛋白HCN2及HCN4。单细胞膜片钳实验检测出现连续性自发性动作电位，证实此种方法可以构建组织工程窦房结的种子细胞。目前组织工程窦房结研究的种子细胞种类多样，主要的方法不外乎用诱导分化、基因修饰的方法将细胞诱导为起搏细胞，但安全性仍没有统一规范的评价。普遍认为种子细胞应来源广泛，取材方便，可以大规模批量提取，且生存能力强而不具备成瘤的风险。组织工程支架材料更是种类繁多，活体的窦房结本身就处于胶原支架上[8]，因此胶原类材料更为适用于组织工程窦房结(图1)。胶原类材料中含有大量的细胞外基质，细胞外基质可以将心肌细胞排列，使浦肯野纤维排列其中，起到传导的作用[9]。胶原类材料中的细胞外基质就较为丰富，Pen?a等[10]在研究心力衰竭时使用了一种高分子仿生聚合物，反向热导凝胶(reverse thermal gel，RTG)，这种凝胶的特殊性在于能够持续性促进心肌细胞的分裂，并且可以作为运载心肌细胞的载体，根据温度将其运送到机体的不同部位。将反向热导凝胶和多聚赖氨酸、层粘蛋白混合进行处理后，能够极大的提高新生及成年大鼠心肌细胞的存活，窦房结的起搏细胞本质上类似于心肌细胞，因此反向热导凝胶不失为一种良好的支架材料。纳米级材料在心脏组织工程中应用也极其广泛。纳米级材料的优势在于能够参与微环境中的信号传导，决定种子细胞的命运，例如促进细胞的诱导分化[11]。Wu等[12]利用碳纳米角作为支架材料能够促进大鼠心肌的存活，同时减少成纤维细胞的形成。而石墨烯作为一种新兴的纳米材料也已经广泛应用于药物输送[13]、细胞成像以及组织支架[14-15]。Shin等[16]将纳米级材料和凝胶混合使用，以氧化还原的石墨烯整合明胶甲基丙烯酰氯水凝胶作为一种新的支架材料，这种支架材料不仅能够促进细胞的增殖、分裂和成熟，还能增强心肌细胞的收缩力，加快其收缩搏动的速率，其降解产物也不会对机体产生不良反应，完全适用与组织工程窦房结。组织工程窦房结不仅需要起搏细胞的自发搏动，还需要移行细胞等进行传导，这种新支架无疑是一种较好的选择。"

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