TUBITAK, Marmara Research Center, Genetic Engineering and Biotechnology Institute, Kocaeli, Turkey.
Istanbul Technical University, Molecular Biology-Genetics and Biotechnology Program - Ayazaga Campus, Istanbul, Turkey.
J Biomater Appl. 2021 Oct;36(4):740-753. doi: 10.1177/08853282211018529. Epub 2021 May 26.
Cardiac tissue engineering focusing on biomaterial scaffolds incorporating cells from different sources has been explored to regenerate or repair damaged area as a lifesaving approach.The aim of this study was to evaluate the cardiomyocyte differentiation potential of human adipose mesenchymal stem cells (hAD-MSCs) as an alternative cell source on silk fibroin (SF) scaffolds for cardiac tissue engineering. The change in surface morphology of SF scaffolds depending on SF concentration (1-6%, w/v) and increase in their porosity upon application of unidirectional freezing were visualized by scanning electron microscopy (SEM). Swelling ratio was found to increase 2.4 fold when SF amount was decreased from 4% to 2%. To avoid excessive swelling, 4% SF scaffold with swelling ratio of 10% (w/w) was chosen for further studies.Biodegradation rate of SF scaffolds depended on enzymatic activity was found to be 75% weight loss of SF scaffolds at the day 14. The phenotype of hAD-MSCs and their multi-linage potential into chondrocytes, osteocytes, and adipocytes were shown by flow cytometry and immunohistochemical staining, respectively.The viability of hAD-MSCs on 3D SF scaffolds was determined as 90%, 118%, and 138% after 1, 7, and 14 days, respectively. The use of 3D SF scaffolds was associated with increased production of cardiomyogenic biomarkers: α-actinin, troponin I, connexin 43, and myosin heavy chain. The fabricated 3D SF scaffolds were proved to sustain hAD-MSCs proliferation and cardiomyogenic differentiation therefore, hAD-MSCs on 3D SF scaffolds may useful tool to regenerate or repair damaged area using cardiac tissue engineering techniques.
以不同来源的细胞为核心，专注于生物材料支架的心脏组织工程已被探索用于再生或修复受损区域，作为一种救生方法。本研究旨在评估人脂肪间充质干细胞（hAD-MSCs）作为替代细胞源在丝素（SF）支架上用于心脏组织工程的心肌细胞分化潜力。通过扫描电子显微镜（SEM）观察 SF 支架的表面形态随 SF 浓度（1-6%，w/v）的变化以及单向冷冻应用时其孔隙率的增加。发现 SF 量从 4%减少到 2%时，溶胀比增加了 2.4 倍。为了避免过度溶胀，选择溶胀比为 10%（w/w）的 4%SF 支架进行进一步研究。SF 支架的降解速率取决于酶活性，发现 SF 支架在第 14 天的重量损失为 75%。通过流式细胞术和免疫组织化学染色分别显示 hAD-MSCs 的表型及其向软骨细胞、成骨细胞和脂肪细胞的多谱系分化潜能。hAD-MSCs 在 3D SF 支架上的活力分别为第 1、7 和 14 天的 90%、118%和 138%。使用 3D SF 支架与心肌生成生物标志物的产生增加有关：α-辅肌动蛋白、肌钙蛋白 I、连接蛋白 43 和肌球蛋白重链。所制造的 3D SF 支架被证明能够维持 hAD-MSCs 的增殖和心肌分化，因此，3D SF 支架上的 hAD-MSCs 可能是使用心脏组织工程技术再生或修复受损区域的有用工具。
