Department of Gastroenterology and Hepatology, General Hospital, Tianjin Medical University, Tianjin, 300052, China.
The Postdoctoral Research Station, School of Medicine, Nankai University, Tianjin, 300071, China.
Stem Cell Rev Rep. 2020 Dec;16(6):1292-1304. doi: 10.1007/s12015-020-10053-2. Epub 2020 Oct 3.
Crohn's disease (CD) with externally fistulizing openings indicates the aggressive and relapsing manifestation and results in undesirable long-term outcomes of patients. MSC-based approach combined with multidisciplinary strategy has mandated a redefinition of the administration and management of numerous recurrent and refractory diseases whereas the spatio-temporal evaluation of the metabolokinetics and efficacy of MSCs on intractable CD with enterocutaneous fistula (EF) are largely inaccessible and dauntingly complex. Herein, we primitively established dual-fluorescence expressing placenta-derived MSCs (DF-MSCs) and explored their multidimensional attributes, including cytomorphology, immunophenotying, multilineage differentiation and long-term proliferation, together with the recognition of bifluorescence intensity (BLI). Then, with the aid of in vivo living imaging, clinicopathological or inflammatory cytokine examinations and in vitro analyses, we systematically and meticulously dissected the metabolokinetics and curative effect of MSCs on mice with refractory Crohn's-like EF (EF mice), together with revealing the underlying mechanism including reactive oxygen species (ROS) and neovascularization. Strikingly, the DF-MSCs exhibited stabilized BLI and biological properties. The spatio-temporal distribution and therapeutic process of MSCs in EF mice were intuitively delineated. Meanwhile, our data indicated the curative mechanisms of DF-MSCs by simultaneously downregulating ROS and accelerating neovascularization. Collectively, we systematically illuminated the spatio-temporal biofunction and mechanism of DF-MSCs on EF mice. Our findings have supplied new references for safety and effectiveness assessments as well as the establishment of guidelines for optimal administrations of MSC-based cytotherapy in preclinical studies, which collectively indicates the prospect of P-MSC administration in clinical trials during a wide spectrum of disease remodeling including the fistulizing CD. Graphical abstract.
克罗恩病（CD）伴外部瘘管开口表明其具有侵袭性和复发性表现，导致患者出现不良的长期结局。基于间充质干细胞（MSC）的方法结合多学科策略，需要重新定义对许多复发性和难治性疾病的管理，而对难治性 CD 伴肠皮瘘（EF）的 MSC 代谢动力学和疗效的时空评估在很大程度上是难以实现和复杂的。在此，我们原始地建立了双荧光表达胎盘来源的间充质干细胞（DF-MSCs），并探索了它们的多维特性，包括细胞形态、免疫表型、多能分化和长期增殖，以及双荧光强度（BLI）的识别。然后，借助体内活体成像、临床病理或炎症细胞因子检查和体外分析，我们系统和细致地剖析了 MSC 对难治性 CD 样 EF（EF 小鼠）的代谢动力学和疗效，同时揭示了包括活性氧（ROS）和新血管生成在内的潜在机制。引人注目的是，DF-MSCs 表现出稳定的 BLI 和生物学特性。DF-MSCs 在 EF 小鼠中的时空分布和治疗过程被直观地描绘出来。同时，我们的数据表明，DF-MSCs 通过同时下调 ROS 和加速新血管生成来发挥治疗作用。总之，我们系统地阐明了 DF-MSCs 对 EF 小鼠的时空生物功能和机制。我们的发现为临床前研究中基于 MSC 的细胞治疗的安全性和有效性评估以及最佳管理指南的建立提供了新的参考，这共同表明了在包括瘘管性 CD 在内的广泛疾病重塑过程中，P-MSC 给药在临床试验中的前景。
