Division of Cardiac Surgery, University of Ottawa Heart Institute, 40 Ruskin Street, Ottawa, ON K1Y 4W7, Canada.
Eur Cell Mater. 2012 Sep 12;24:175-95; discussion 195-6. doi: 10.22203/ecm.v024a13.
Biomaterial-guided regeneration represents a novel approach for the treatment of myopathies. Revascularisation and the intramuscular extracellular matrix are important factors in stimulating myogenesis and regenerating muscle damaged by ischaemia. In this study, we used an injectable collagen matrix, enhanced with sialyl LewisX (sLeX), to guide skeletal muscle differentiation and regeneration. The elastic properties of collagen and sLeX-collagen matrices were similar to those of skeletal muscle, and culture of pluripotent mESCs on the matrices promoted their differentiation into myocyte-like cells expressing Pax3, MHC3, myogenin and Myf5. The regenerative properties of matrices were evaluated in ischaemic mouse hind-limbs. Treatment with the sLeX-matrix augmented the production of myogenic-mediated factors insulin-like growth factor (IGF)-1, and IGF binding protein-2 and -5 after 3 days. This was followed by muscle regeneration, including a greater number of regenerating myofibres and increased transcription of Six1, M-cadherin, myogenin and Myf5 after 10 days. Simultaneously, the sLeX-matrix promoted increased mobilisation and engraftment of bone marrow-derived progenitor cells, the development of larger arterioles and the restoration of tissue perfusion. Both matrix treatments tended to reduce maximal forces of ischaemic solei muscles, but sLeX-matrix lessened this loss of force and also prevented muscle fatigue. Only sLeX-matrix treatment improved mobility of mice on a treadmill. Together, these results suggest a novel approach for regenerative myogenesis, whereby treatment only with a matrix, which possesses an inherent ability to guide myogenic differentiation of pluripotent stem cells, can enhance the endogenous vascular and myogenic regeneration of skeletal muscle, thus holding promise for future clinical use.
生物材料引导的再生代表了一种治疗肌病的新方法。再血管化和肌内细胞外基质是刺激成肌发生和再生缺血性损伤肌肉的重要因素。在这项研究中，我们使用了一种可注射的胶原蛋白基质，该基质增强了唾液酸化 LewisX（sLeX），以指导骨骼肌分化和再生。胶原蛋白和 sLeX-胶原蛋白基质的弹性特性与骨骼肌相似，并且在基质上培养多能 mESCs 可促进其分化为表达 Pax3、MHC3、myogenin 和 Myf5 的肌细胞样细胞。在缺血性小鼠后肢中评估了基质的再生特性。用 sLeX-基质处理可在 3 天后增加肌生成介导因子胰岛素样生长因子 (IGF)-1 和 IGF 结合蛋白-2 和 -5 的产生。随后进行肌肉再生，包括 10 天后再生肌纤维数量增加和 Six1、M-cadherin、myogenin 和 Myf5 的转录增加。同时，sLeX-基质促进了骨髓源性祖细胞的动员和植入，较大的小动脉的发育以及组织灌注的恢复。两种基质处理均倾向于降低缺血性比目鱼肌的最大力，但 sLeX-基质减轻了这种力的丧失，并且还防止了肌肉疲劳。只有 sLeX-基质处理可改善跑步机上小鼠的运动能力。总之，这些结果表明了一种新的再生肌发生方法，即仅用具有固有能力指导多能干细胞成肌分化的基质进行治疗，可以增强骨骼肌的内源性血管和肌生成再生，因此有望在未来的临床应用中使用。
