Department of Orthopaedic Surgery, Sir Run Run Shaw Hospital, Medical College of Zhejiang University, Hangzhou, China.
Department of Orthopaedic Surgery, Sir Run Run Shaw Hospital, Medical College of Zhejiang University, Hangzhou, China.
Acta Biomater. 2018 Mar 1;68:15-28. doi: 10.1016/j.actbio.2017.12.035. Epub 2017 Dec 30.
Muscle injury and defect affect people's quality of life, and effective treatment is lacking. Herein, we generated a scaffold to obtain decellularized porcine Achilles tendon myotendinous junction (D-MTJ) extracellular matrix (ECM) with well-preserved native biphasic hierarchical structure, biological composition, and excellent mechanical properties for muscle regeneration. The combined use of potassium chloride, potassium iodide, Triton-X 100, and sodium-dodecyl sulfate (SDS) can completely remove the main immunogenicity, while maintaining the major biological components and microstructure. The specific biomechanics of D-MTJ is comparable to the native muscle-tendon physiological conditions. Additionally, the D-MTJ ECM scaffold induced minimal immunological reaction (histology analysis) through rat subcutaneous implantation. Moreover, in vitro, muscle satellite cells adhered, proliferated, and infiltrated into the D-MTJ scaffold, and myofiber-like cell differentiation was observed as shown by increased expression of myogenesis-related genes during culture. In vivo, newly formed myofibers were observed in a muscle defect model with D-MTJ orthotopic transplantation, while the control group presented mostly with fibrous tissue deposition. Additionally, the number of Myod and MyHC-positive cells in the ECM scaffold group was higher at day 30. We preliminary explored the mechanisms underlying D-MTJ-mediated muscle regeneration, which may be attributed to its specific biphasic hierarchical structure, bio-components, and attractiveness for myogenesis cells. In conclusion, our findings suggest the D-MTJ ECM scaffold prepared in this study is a promising choice for muscle regeneration.
This study is the first to use decellularization technology obtaining the specifically decellularized myotendinous junction (D-MTJ) with well-preserved biphasic hierarchical structure and constituents, excellent mechanical properties and good biocompatibility. The D-MTJ was further proved to be efficient for muscle regeneration in vitro and in vivo, and the underlying mechanisms may be attributed to its specifically structure and constituents, improved myogenesis and good preservation of repair-related factors. Our study may provide basis for the decellularization of other biphasic hierarchical tissues and a platform for further studies on muscle fiber and tendon integrations in vitro.
肌肉损伤和缺陷会影响人们的生活质量，而目前缺乏有效的治疗方法。在此，我们生成了一种支架，以获得具有保留完好的天然双相分层结构、生物组成和优异机械性能的去细胞猪跟腱肌腱结合部（D-MTJ）细胞外基质（ECM），用于肌肉再生。联合使用氯化钾、碘化钾、Triton-X100 和十二烷基硫酸钠（SDS）可以完全去除主要的免疫原性，同时保持主要的生物成分和微观结构。D-MTJ 的特定生物力学与天然肌肉-肌腱生理条件相当。此外，通过大鼠皮下植入，D-MTJ ECM 支架引起的免疫反应最小（组织学分析）。此外，在体外，肌肉卫星细胞黏附、增殖并渗透到 D-MTJ 支架中，并且在培养过程中观察到肌生成相关基因表达增加，表明肌纤维样细胞分化。在体内，在 D-MTJ 原位移植的肌肉缺陷模型中观察到新形成的肌纤维，而对照组主要表现为纤维组织沉积。此外，在第 30 天时，ECM 支架组中的 Myod 和 MyHC 阳性细胞数量更高。我们初步探讨了 D-MTJ 介导的肌肉再生的机制，这可能归因于其特定的双相分层结构、生物成分以及对肌生成细胞的吸引力。总之，我们的研究结果表明，本研究中制备的 D-MTJ ECM 支架是肌肉再生的有前途的选择。
本研究首次使用去细胞化技术获得具有保留完好的双相分层结构和成分、优异的机械性能和良好的生物相容性的特异性去细胞化肌腱结合部（D-MTJ）。D-MTJ 进一步证明在体外和体内均能有效促进肌肉再生，其潜在机制可能归因于其特定的结构和成分、改善的肌生成以及修复相关因子的良好保留。我们的研究可能为其他双相分层组织的去细胞化提供基础，并为进一步研究体外肌纤维和肌腱整合提供平台。
