Center for Human Tissues & Organs Degeneration, Shenzhen Institute of Advanced Technology, Chinese Academy of Sciences, Shenzhen, Guangdong 518055, China; Center for Biomaterials, Korea Institute of Science and Technology (KIST), Seoul 02792, Republic of Korea.
Center for Biomaterials, Korea Institute of Science and Technology (KIST), Seoul 02792, Republic of Korea.
Mater Sci Eng C Mater Biol Appl. 2020 Aug;113:110961. doi: 10.1016/j.msec.2020.110961. Epub 2020 Apr 19.
Decellularized human lung fibroblast-derived matrix (hFDM) has demonstrated its excellent proangiogenic capability. In this study, we propose a self-assembled, injectable multicellular microspheres containing human umbilical vein endothelial cells (HUVECs) and mesenchymal stem cell (MSCs), collagen hydrogel (Col), and hFDM toward therapeutic angiogenesis. Those multicellular microspheres are spontaneously formed by the mixtures of cell and hydrogel after being dropped on the parafilm for several hours. The size of microspheres can be manipulated via adjusting the initial volume of droplets and the culture period. The cells in the microspheres are highly viable. Multicellular microspheres show good capability of cell migration on 2D culture plate and also exhibit active cell sprouting in 3D environment (Col) forming capillary-like structures. We also find that multiple angiogenic-related factors are significantly upregulated with the multicellular microspheres prepared via Col and hFDM (Col/hFDM) than those prepared using Col alone or single cells (harvested from cocultured HUVECs/MSCs in monolayer). For therapeutic efficacy evaluation, three different groups of single cells, Col and Col/hFDM microspheres are injected to a hindlimb ischemic model, respectively, along with PBS injection as a control group. It is notable that Col/hFDM microspheres significantly improve the blood reperfusion and greatly attenuate the fibrosis level of the ischemic regions. In addition, Col/hFDM microspheres show higher cell engraftment level than that of the other groups. The incorporation of transplanted cells with host vasculature is detectable only with the treatment of Col/hFDM. Current results suggest that hFDM plays an important role in the multicellular microspheres for angiogenic cellular functions in vitro as well as in vivo. Taken together, our injectable multicellular microspheres (Col/hFDM) offer a very promising platform for cell delivery and tissue regenerative applications.
脱细胞人肺成纤维细胞衍生基质（hFDM）已证明具有出色的促血管生成能力。在这项研究中，我们提出了一种自组装的、可注射的含有人脐静脉内皮细胞（HUVEC）和间充质干细胞（MSCs）、胶原水凝胶（Col）和 hFDM 的多细胞微球，用于治疗性血管生成。这些多细胞微球是通过将细胞和水凝胶的混合物滴在 Parafilm 上数小时后自发形成的。通过调整初始液滴体积和培养时间，可以控制微球的大小。微球中的细胞具有很高的活力。多细胞微球在 2D 培养板上显示出良好的细胞迁移能力，并且在 3D 环境（Col）中也表现出活跃的细胞发芽，形成类似毛细血管的结构。我们还发现，与单独使用 Col 或单细胞（从共培养的 HUVECs/MSCs 单层中收获）制备的微球相比，使用 Col 和 hFDM 制备的多细胞微球显著上调了多种血管生成相关因子。为了评估治疗效果，将三种不同的细胞群、Col 和 Col/hFDM 微球分别注射到后肢缺血模型中，以 PBS 注射作为对照组。值得注意的是，Col/hFDM 微球显著改善了血液再灌注，并大大减轻了缺血区域的纤维化程度。此外，Col/hFDM 微球的细胞植入水平高于其他组。只有在用 Col/hFDM 处理时，才能检测到移植细胞与宿主血管的整合。目前的结果表明，hFDM 在体外和体内多细胞微球的血管生成细胞功能中起着重要作用。总之，我们的可注射多细胞微球（Col/hFDM）为细胞递送和组织再生应用提供了一个非常有前途的平台。
