Department of Animal Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran.
Department of Animal Science, College of Agriculture, Isfahan University of Technology, Isfahan, Iran.
Cell Biochem Funct. 2022 Oct;40(7):668-682. doi: 10.1002/cbf.3728. Epub 2022 Aug 4.
Adipose-derived mesenchymal stem cells (Ad-MSCs) are promising candidates for cardiac repair/regeneration. The application of copolymer nanoscaffolds has received great attention in tissue engineering to support differentiation and functional tissue organization toward effective tissue regeneration. The objective of the current study was to develop functional and bioactive scaffolds by combining polycaprolactone (PCL) and polylactic acid (PLA) for cardiomyocyte differentiation of human Ad-MSC (hAd-MSCs) in the absence or presence of 5-azacytidine and transforming growth factor-β (TGF-β). To that end, the human MSCs were extracted from human adipose tissue (AD). The cardiomyocyte differentiation potency of hAd-MSCs was evaluated on the novel synthetic PCL/PLA nanofiber scaffolds prepared in the absence and presence of 5-azacytidine and TGF-β supplements. A PCL/PLA nanofibrous scaffold was fabricated using the electrospinning method and its nanotopography and porous structure were characterized using scanning electron microscopy. In addition, the attachment of hAd-MSCs on the PCL/PLA scaffolds was semiquantitatively investigated. Compared with other treatments, the PCL/PLA nanofibrous scaffold supplemented with both 5-azacytidine and TGF-β was observed to differentiate hAd-MSCs into cardiomyocytes at Day 21 as evidenced by real-time PCR for cardiac-specific genes including cardiac troponin I (cTnI), GATA4, MYH7, and NKX2.5. In addition, flow cytometric analysis of cTnI-positive cells demonstrated that the cardiomyocyte differentiation of hAd-MSCs was more efficient on the PCL/PLA nanofibrous scaffold supplemented with both 5-azacytidine and TGF-β than it was in the other treatment groups. Generally speaking, the results show that PCL/PLA nanofibrous scaffolds may be applied as a platform for efficient differentiation of hAd-MSCs into functional cardiomyocytes.
脂肪间充质干细胞（Ad-MSCs）是心脏修复/再生的有前途的候选物。共聚物纳米支架在组织工程中得到了广泛关注，以支持分化和功能性组织组织，从而实现有效的组织再生。本研究的目的是通过结合聚己内酯（PCL）和聚乳酸（PLA）来开发功能性和生物活性支架，以促进人脂肪间充质干细胞（hAd-MSCs）向心肌细胞分化，同时存在或不存在 5-氮杂胞苷和转化生长因子-β（TGF-β）。为此，从人脂肪组织（AD）中提取人 MSCs。在不存在和存在 5-氮杂胞苷和 TGF-β补充剂的情况下，评估 hAd-MSCs 在新型合成 PCL/PLA 纳米纤维支架上的心肌细胞分化能力。使用静电纺丝法制备 PCL/PLA 纳米纤维支架，并使用扫描电子显微镜对其纳米形貌和多孔结构进行表征。此外，还对半定量研究了 hAd-MSCs 在 PCL/PLA 支架上的附着情况。与其他处理相比，在补充有 5-氮杂胞苷和 TGF-β的 PCL/PLA 纳米纤维支架上观察到 hAd-MSCs 在第 21 天分化为心肌细胞，这表现为实时 PCR 检测到包括心脏肌钙蛋白 I（cTnI）、GATA4、MYH7 和 NKX2.5 在内的心脏特异性基因。此外，cTnI 阳性细胞的流式细胞术分析表明，在补充有 5-氮杂胞苷和 TGF-β的 PCL/PLA 纳米纤维支架上，hAd-MSCs 的心肌细胞分化效率高于其他处理组。总的来说，结果表明 PCL/PLA 纳米纤维支架可作为高效分化 hAd-MSCs 为功能性心肌细胞的平台。
