Tenogenic adipose-derived stem cell sheets with nanoyarn scaffolds for tendon regeneration
Tenogenic adipose-derived stem cell sheets with nanoyarn scaffolds for tendon regeneration
复制标题
具有纳米纱线支架的肌腱脂肪干细胞片用于肌腱再生
DOI：
10.1016/j.msec.2020.111506
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发表时间：
2021-02-01
影响因子：
7.9
通讯作者：
Fan, Cunyi
中科院分区：
文献类型：
--
作者：
Chen, Shuai;Wang, Juan;Fan, Cunyi
组织工程，特别是基于细胞片的工程，为肌腱再生提供了一种有前途的方法。然而，获得足够的细胞来源用于组织工程应用具有挑战性。脂肪干细胞 (ASC) 是组织再生的重要来源，并已被证明具有通过生长分化因子 5 (GDF-5) 诱导进行体外肌腱分化的潜力。在这项研究中，我们探讨了 GDF-5 刺激的 ASC 细胞片用于工程肌腱修复的可行性。定量聚合酶链反应和蛋白质印迹显示，与未诱导的相比，GDF-5 诱导的 ASC 细胞片层中肌腱形成相关标记物（Col I&III、TNMD、双糖链蛋白聚糖和腱蛋白 C）显着增加。此外，SMAD2/3蛋白和磷酸化SMAD1/5/9的水平显着增强，表明GDF-5可能通过SMAD1/5/9的磷酸化发挥其功能。此外，细胞片层与 P(LLA-CL)/丝素蛋白纳米纱支架相结合，形成肌腱组织工程的构建体。末端脱氧核苷酸转移酶 dUTP 缺口末端标记和免疫荧光测定表明 GDF-5 诱导的构建体中具有良好的细胞活力和肌腱发生相关标记物表达。此外，组织学、免疫组织化学和生物力学分析证明，该结构在兔模型中显示出肌腱修复的潜力。在我们的研究中，我们通过GDF-5诱导的ASC细胞片和纳米纱线支架的结合，成功生产了一种新的组织工程肌腱，这对肌腱再生很有价值。
Tissue engineering, especially cell sheets-based engineering, offers a promising approach to tendon regeneration; however, obtaining a sufficient source of cells for tissue engineering applications is challenging. Adipose-derived stem cells (ASCs) are essential sources for tissue regeneration and have been shown to have the potential for tenogenic differentiation in vitro via induction by growth differentiation factor 5 (GDF-5). In this study, we explored the feasibility of ASCs cell sheets stimulated by GDF-5 for engineered tendon repair. As shown by quantitative polymerase chain reaction and western blotting, tenogenesis-related markers (Col I&III, TNMD, biglycan, and tenascin C) were significantly increased in GDF-5-induced ASCs cell sheets compared with the uninduced. Moreover, the levels of SMAD2/3 proteins and phospho-SMAD1/5/9 were significantly enhanced, demonstrating that GDF-5 may exert its functions through phosphorylation of SMAD1/5/9. Furthermore, the cell sheets were combined with P(LLA-CL)/Silk fibroin nanoyarn scaffolds to form constructs for tendon tissue engineering. Terminal deoxynucleotidyl transferase dUTP nick end labeling and immunofluorescence assays demonstrated favorable cell viability and tenogenesis-related marker expression in GDF-5-induced constructs. In addition, the constructs showed the potential for tendon repair in rabbit models, as demonstrated by histological, immunohistochemical, and biomechanical analyses. In our study, we successfully produced a new tissue-engineered tendon by the combination of GDF-5-induced ASCs cell sheets and nanoyarn scaffold which is valuable for tendon regeneration.