Institute of Pharmacology and Toxicology, University Medical Center Goettingen, Germany; Randall Centre for Cell and Molecular Biophysics, King's College London, London, UK; DZHK (German Center for Cardiovascular Research) partner site, Goettingen, Germany.
Institute of Pharmacology and Toxicology, University Medical Center Goettingen, Germany; DZHK (German Center for Cardiovascular Research) partner site, Goettingen, Germany.
Biomater Adv. 2022 Aug;139:213041. doi: 10.1016/j.bioadv.2022.213041. Epub 2022 Jul 21.
Tissue engineering with human cardiac fibroblasts (CF) allows identifying novel mechanisms and anti-fibrotic drugs in the context of cardiac fibrosis. However, substantial knowledge on the influences of the used materials and tissue geometries on tissue properties and cell phenotypes is necessary to be able to choose an appropriate model for a specific research question. As there is a clear lack of information on how CF react to the mold architecture in engineered connective tissues (ECT), we first compared the effect of two mold geometries and materials with different hardnesses on the biomechanical properties of ECT. We could show that ECT, which formed around two distant poles (non-uniform model) were less stiff and more strain-resistant than ECT, which formed around a central rod (uniform model), independent of the materials used for poles and rods. Next, we investigated the cell state and could demonstrate that in the uniform versus non-uniform model, the embedded cells have a higher cell cycle activity and display a more pronounced myofibroblast phenotype. Differential gene expression analysis revealed that uniform ECT displayed a fibrosis-associated gene signature similar to the diseased heart. Furthermore, we were able to identify important relationships between cell and tissue characteristics, as well as between biomechanical tissue parameters by implementing cells from normal heart and end-stage heart failure explants from patients with ischemic or dilated cardiomyopathy. Finally, we show that the application of pro- and anti-fibrotic factors in the non-uniform and uniform model, respectively, is not sufficient to mimic the effect of the other geometry. Taken together, we demonstrate that modifying the mold geometry in tissue engineering with CF offers the possibility to compare different cellular phenotypes and biomechanical tissue properties.
组织工程用人心脏成纤维细胞（CF）可用于确定心脏纤维化背景下的新机制和抗纤维化药物。然而，为了能够选择合适的模型来回答特定的研究问题，需要了解用于组织的材料和组织几何形状对组织特性和细胞表型的影响。由于在工程化结缔组织（ECT）中 CF 对模具结构的反应信息非常有限，因此我们首先比较了两种模具几何形状和不同硬度的材料对 ECT 生物力学特性的影响。结果表明，围绕两个远极点形成的 ECT（非均匀模型）比围绕中心杆形成的 ECT（均匀模型）更柔软，更能抵抗应变，而与使用的材料无关。接下来，我们研究了细胞状态，并表明在均匀模型与非均匀模型中，嵌入式细胞的细胞周期活性更高，呈现出更明显的肌成纤维细胞表型。差异基因表达分析显示，均匀 ECT 显示出与疾病心脏相似的纤维化相关基因特征。此外，我们通过应用来自正常心脏和缺血性或扩张型心肌病患者的心衰末期心脏组织的细胞，成功地鉴定了细胞和组织特性之间以及生物力学组织参数之间的重要关系。最后，我们证明在非均匀模型和均匀模型中分别应用促纤维化和抗纤维化因子不足以模拟另一种几何形状的效果。总之，我们证明了在 CF 组织工程中修改模具几何形状可以比较不同的细胞表型和生物力学组织特性。
