Laboratory of Nano-regenerative Medicine, Department of Biomedical Science, College of Life Science, CHA University, CHA Biocomplex, 335 Pangyo-ro, Sampyeong-Dong, Bundang-gu, Seongnam-si, 13488, Republic of Korea.
Laboratory of Nano-regenerative Medicine, Department of Biomedical Science, College of Life Science, CHA University, CHA Biocomplex, 335 Pangyo-ro, Sampyeong-Dong, Bundang-gu, Seongnam-si, 13488, Republic of Korea.
Biomaterials. 2023 Nov;302:122350. doi: 10.1016/j.biomaterials.2023.122350. Epub 2023 Oct 15.
Many attempts have been made to use mitochondria (MT) to treat human diseases; however, MT are large, making them difficult to deliver effectively. Therefore, a transfer strategy based on membrane fusion was established. Fusogenic mitochondrial capsules (FMCs) comprising a neutral lipid (PE), a cationic lipid (DOTAP), an aromatic lipid (Liss Rhod PE), and three types of liposome (FMC0, FMC1, and FMC2), were designed and synthesized. The amount of DOTAP, which affects membrane fusion efficiency, differed between FMC preparations. The characteristics of these FMCs were analyzed by DLS, TEM, and AFM, and the encapsulation and fusion efficiency between FMC-MT and FMC-chondrocytes were confirmed by FRET, mtDNA copy number, and CLSM, respectively. Compared with naked MT, delivery of FMCs to chondrocytes was faster and more efficient. Moreover, fusion was a more stable delivery method than endocytosis, as evidenced by reduced induction of mitophagy. In vitro and in vivo experiments revealed that FMCs reduced expression of inflammatory cytokines and MMP13, increased expression of extracellular matrix components, and promoted cartilage regeneration. These findings suggest that FMCs are a highly effective and promising strategy for delivery of MT to promote cartilage regeneration, and highlight their potential as a novel platform for MT transfer therapy.
已经有许多尝试利用线粒体（MT）来治疗人类疾病的方法；然而，MT 较大，使得它们难以有效传递。因此，建立了一种基于膜融合的传递策略。设计并合成了由中性脂质（PE）、阳离子脂质（DOTAP）、芳香脂质（Liss Rhod PE）和三种脂质体（FMC0、FMC1 和 FMC2）组成的融合线粒体胶囊（FMC）。DOTAP 的量会影响膜融合效率，因此不同 FMC 制剂之间存在差异。通过 DLS、TEM 和 AFM 分析这些 FMC 的特性，并通过 FRET、mtDNA 拷贝数和 CLSM 分别确认 FMC-MT 和 FMC-软骨细胞之间的封装和融合效率。与裸 MT 相比，FMC 向软骨细胞的传递更快、更有效。此外，融合是一种比内吞作用更稳定的传递方法，这可以从减少诱导的线粒体自噬得到证明。体外和体内实验表明，FMC 可降低炎症细胞因子和 MMP13 的表达，增加细胞外基质成分的表达，并促进软骨再生。这些发现表明，FMC 是一种高效、有前途的传递 MT 以促进软骨再生的策略，并突出了它们作为 MT 传递治疗的新型平台的潜力。
