1 Leibniz Research Laboratories for Biotechnology and Artificial Organs (LEBAO), Department of Cardiothoracic, Transplantation and Vascular Surgery, Hannover Medical School , Hannover, Germany .
Tissue Eng Part A. 2014 Feb;20(3-4):799-809. doi: 10.1089/ten.TEA.2013.0184. Epub 2013 Nov 16.
The in vitro generation of a bioartificial cardiac construct (CC) represents a promising tool for the repair of ischemic heart tissue. Several approaches to engineer cardiac tissue in vitro have been conducted. The main drawback of these studies is the insufficient size of the resulting construct for clinical applications. The focus of this study was the generation of an artificial three-dimensional (3D), contractile, and suturable myocardial patch by combining a gel-based CC with decellularized porcine small intestinal submucosa (SIS), thereby engineering an artificial tissue of 11 cm² in size. The alignment and morphology of rat neonatal cardiomyocytes (rCMs) in SIS-CC complexes were investigated as well as the re-organization of primary endothelial cells which were co-isolated in the rCM preparation. The ability of a rat heart endothelial cell line (RHE-A) to re-cellularize pre-existing vessel structures within the SIS or a biological vascularized matrix (BioVaM) was determined. SIS-CC contracted spontaneously, uniformly, and rhythmically with an average rate of 200 beats/min in contrast to undirected contractions observed in CC without SIS support. rCM exhibited an elongated morphology with well-defined sarcomeric structures oriented along the longitudinal axis in the SIS-CC, whereas round-shaped and random-arranged rCM were observed in CC. Electric coupling of rCM was demonstrated by microelectrode array measurements. A dense network of CD31⁺/eNOS⁺ cells was detected as permeating the whole construct. Superficial supplementation of RHE-A cells to SIS-CC led to the migration of these cells through the CC, resulting in the re-population of pre-existing vessel structures within the decelluarized SIS. By infusion of RHE-A cells into the BioVaM venous and arterial pedicles, a re-population of the BioVaM vessel bed as well as distribution of RHE-A cells throughout the CC was achieved. Rat endothelial cells within the CC were in contact with RHE-A cells. Ingrowth and formation of a network by endothelial cells infused through the BioVaM represent a promising step toward engineering a functional perfusion system, enabling the engineering of vascularized and well-nourished 3D CC of dimensions relevant for therapeutic heart repair.
体外生成生物人工心脏构建体（CC）代表了修复缺血性心脏组织的一种很有前途的工具。已经有几种方法可以在体外构建心脏组织。这些研究的主要缺点是所得构建体的尺寸不足以用于临床应用。本研究的重点是通过将基于凝胶的 CC 与脱细胞化的猪小肠黏膜下层（SIS）结合来生成一个人工三维（3D）、收缩性和可缝合的心肌贴片，从而构建一个 11cm²大小的人工组织。研究了大鼠新生心肌细胞（rCM）在 SIS-CC 复合物中的排列和形态，以及在 rCM 制备中共同分离的原代内皮细胞的重新组织。确定了大鼠心脏内皮细胞系（RHE-A）在 SIS 或生物血管化基质（BioVaM）内重新填充现有血管结构的能力。SIS-CC 自发、均匀且有节奏地收缩，平均速率为 200 次/分钟，与没有 SIS 支持的 CC 中观察到的无定向收缩形成对比。rCM 表现出伸长的形态，具有沿着 SIS-CC 的纵轴取向的定义明确的肌节结构，而在 CC 中观察到圆形和随机排列的 rCM。通过微电极阵列测量证明了 rCM 的电耦合。检测到 CD31⁺/eNOS⁺细胞密集网络贯穿整个构建体。将 RHE-A 细胞浅层补充到 SIS-CC 中，导致这些细胞通过 CC 迁移，从而使脱细胞化 SIS 内的现有血管结构重新填充。通过将 RHE-A 细胞注入 BioVaM 的静脉和动脉蒂，实现了 BioVaM 血管床的重新填充以及 RHE-A 细胞在整个 CC 中的分布。CC 内的大鼠内皮细胞与 RHE-A 细胞接触。通过 BioVaM 注入的内皮细胞的向内生长和形成网络是朝着构建功能性灌注系统迈出的有前途的一步，这使得可以构建血管化和营养良好的 3D CC，其尺寸与治疗性心脏修复相关。
