Khakpour Elahe, Tavassoli Amin, Mahdavi-Shahri Nasser, Matin Maryam M
Department of Biology, Faculty of Science, Ferdowsi University of Mashhad, Mashhad, Iran.
Division of Biotechnology, Faculty of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, Iran.
Cell Tissue Bank. 2023 Mar;24(1):11-24. doi: 10.1007/s10561-022-10012-w. Epub 2022 May 21.
Tendon is a collagen-enriched, tough, and intricately arranged connective tissue that connects muscle to the bone and transmits forces, resulting in joint movement. High mechanical demands can affect normal tissues and may lead to severe disorders, which usually require replacement of the damaged tendon. In recent decades, various decellularization methods have been studied for tissue engineering applications. One of the major challenges in tendon decellularization is preservation of the tendon extracellular matrix (ECM) architecture to maintain natural tissue characteristics. The aim of the present study was to create a decellularized bovine Achilles tendon scaffold to investigate its cytocompatibility with seeded hAd-MSCs (human adipose derived-mesenchymal stem cells) and blastema tissue in vitro. Here, we describe a reliable procedure to decellularize bovine Achilles tendon using a combination of physical and chemical treatments including repetitive freeze-thaw cycles and the ionic detergent SDS, respectively. The decellularization effectiveness and cytocompatibility of the tendon scaffolds were verified by histological studies and scanning electron microscopy for up to 30 days after culture. Histological studies revealed hAd-MSC attachment and penetration into the scaffolds at 5, 10, 15 and 20 days of culture. However, a decrease in cell number was observed on days 25 and 30 after culture in vitro. Moreover, migration of the blastema tissue cells into the scaffold were shown at 10 to 25 days post culture, however, destruction of the scaffolds and reduction in cell number were observed on 30th day after culture. Our results suggest that this decellularization protocol is an effective and biocompatible procedure which supports the maintenance and growth of both hAd-MSCs and blastema cells, and thus might be promising for tendon tissue engineering.
肌腱是一种富含胶原蛋白、坚韧且结构复杂的结缔组织，它将肌肉与骨骼相连并传递力量，从而导致关节运动。高机械需求会影响正常组织，并可能导致严重疾病，通常需要替换受损的肌腱。近几十年来，人们对各种脱细胞方法进行了研究，以用于组织工程应用。肌腱脱细胞的主要挑战之一是保留肌腱细胞外基质（ECM）结构，以维持天然组织特性。本研究的目的是创建一种脱细胞牛跟腱支架，以研究其与接种的人脂肪来源间充质干细胞（hAd-MSCs）和芽基组织在体外的细胞相容性。在此，我们描述了一种可靠的程序，分别使用包括重复冻融循环和离子去污剂SDS在内的物理和化学处理组合来使牛跟腱脱细胞。通过组织学研究和扫描电子显微镜对培养长达30天的肌腱支架的脱细胞效果和细胞相容性进行了验证。组织学研究显示，在培养的第5、10、15和20天，hAd-MSCs附着并渗透到支架中。然而，在体外培养第25天和30天时观察到细胞数量减少。此外，在培养后第10至25天显示芽基组织细胞迁移到支架中，然而，在培养第30天时观察到支架破坏和细胞数量减少。我们的结果表明，这种脱细胞方案是一种有效且生物相容的程序，可支持hAd-MSCs和芽基细胞的维持和生长，因此可能在肌腱组织工程中具有前景。
