Kim Jiseong, Lee Myung Chul, Jeon Jieun, Rodríguez-delaRosa Alejandra, Endo Yori, Kim Da-Seul, Madrigal-Salazar Andrea Donaxi, Seo Jeong Wook, Lee Hyeseon, Kim Ki-Tae, Moon Jae-I, Park Seung Gwa, Lopez-Pacheco Mariana Carolina, Alkhateeb Abdulhameed F, Sobahi Nebras, Bassous Nicole, Liu Wenpeng, Lee Jae Seo, Kim Seongsoo, Aykut Dilara Yilmaz, Nasr Mahmoud Lotfi, Hussain Mohammad Asif, Lee Soo-Hong, Kim Woo-Jin, Pourquié Olivier, Sinha Indranil, Shin Su Ryon
Division of Engineering in Medicine, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Cambridge, MA, 02139, USA.
Department of Medical Biotechnology, Dongguk University, 32 Dongguk-ro, Goyang, 10326, Republic of Korea.
Bioact Mater. 2024 Oct 14;43:579-602. doi: 10.1016/j.bioactmat.2024.09.013. eCollection 2025 Jan.
In regenerative medicine, extracellular vesicles (EVs) possess the potential to repair injured cells by delivering modulatory factors. However, the therapeutic effect of EVs in large-scale tissue defects, which are subject to prolonged timelines for tissue architecture and functional restoration, remains poorly understood. In this study, we introduce EVs and cell-tethering hybrid hydrogels composed of tyramine-conjugated gelatin (GelTA) that can be crosslinked with EVs derived from human induced pluripotent stem cell-derived myofibers (hiPSC-myofibers) and hiPSC-muscle precursor cells. This hybrid hydrogel sustains the release of EVs and provides a beneficial nano-topography and mechanical properties for creating a favorable extracellular matrix. Secreted EVs from the hiPSC-myofibers contain specific microRNAs, potentially improving myogenesis and angiogenesis. Herein, we demonstrate increased myogenic markers and fusion/differentiation indexes through the combinatory effects of EVs and integrin-mediated adhesions in the 3D matrix. Furthermore, we observe a unique impact of EVs, which aid in maintaining the viability and phenotype of myofibers under harsh environments. The hybrid hydrogel crosslinked with hiPSCs and EVs is facilely used to fabricate large-scale muscle constructs by the stacking of micro-patterned hydrogel domains. Later, we confirmed a combinational effect, whereby muscle tissue regeneration and functional restoration were improved, via an murine volumetric muscle loss model.
在再生医学中，细胞外囊泡（EVs）具有通过递送调节因子来修复受损细胞的潜力。然而，EVs在大规模组织缺损中的治疗效果仍知之甚少，因为这类缺损的组织结构和功能恢复需要较长时间。在本研究中，我们引入了由酪胺共轭明胶（GelTA）组成的EVs和细胞 tethering 混合水凝胶，其可与源自人诱导多能干细胞衍生的肌纤维（hiPSC-肌纤维）和hiPSC-肌肉前体细胞的EVs交联。这种混合水凝胶可维持EVs的释放，并为创建有利的细胞外基质提供有益的纳米拓扑结构和机械性能。来自hiPSC-肌纤维的分泌型EVs含有特定的微小RNA，可能会改善肌生成和血管生成。在此，我们通过3D基质中EVs和整合素介导的粘附的联合作用，证明了肌生成标志物和融合/分化指数的增加。此外，我们观察到EVs的独特作用，其有助于在恶劣环境下维持肌纤维的活力和表型。与hiPSCs和EVs交联的混合水凝胶可通过堆叠微图案水凝胶结构域轻松用于制造大规模肌肉构建体。随后，我们通过小鼠体积性肌肉损失模型证实了一种联合效应，即肌肉组织再生和功能恢复得到改善。
