Beijing Key Laboratory of Traditional Chinese Medicine Basic Research on Prevention and Treatment for Major Diseases, Experimental Research Center, China Academy of Chinese Medical Sciences, Beijing, China; School of Chemistry and Biological Engineering, University of Science and Technology Beijing, Beijing, China.
School of Chemistry and Biological Engineering, University of Science and Technology Beijing, Beijing, China; Daxing Research Institute, University of Science and Technology Beijing, Beijing, China.
Cytotherapy. 2022 Nov;24(11):1095-1104. doi: 10.1016/j.jcyt.2022.07.003. Epub 2022 Sep 5.
Stem cells from human exfoliated deciduous teeth (SHED) play a significant role in tissue engineering and regenerative medicine. Angiogenesis is crucial in tissue regeneration and a primary target of regenerative medicine. As a first-line anti-diabetic drug, metformin demonstrates numerous valuable impacts on stem cells. This study aimed to explore metformin's impact and mechanism of action on SHED-mediated angiogenesis.
First, cell proliferation; flow cytometry; osteogenic, adipogenic and chondrogenic induction; and proteomics analyses were conducted to explore the role of metformin in SHED. Subsequently, migration and tube formation assays were used to evaluate chemotaxis and angiogenesis enhancement by SHED pre-treated with metformin under co-culture conditions in vitro, and relative messenger RNA expression levels were determined by quantitative reverse transcription polymerase chain reaction. Finally, nude mice were used for in vivo tube formation assay, and sections were analyzed through immunohistochemistry staining with anti-human CD31 antibody.
Metformin significantly promoted SHED proliferation as well as osteogenic, adipogenic and chondrogenic differentiation. Proteomics showed that metformin significantly upregulated 124 differentially abundant proteins involved in intracellular processes, including various proteins involved in cell migration and angiogenesis, such as MAPK1. The co-culture system demonstrated that SHED pre-treated with metformin significantly improved the migration and angiogenesis of human umbilical vein endothelial cells. In addition, SHED pre-treated with metformin possessed greater ability to promote angiogenesis in vivo.
In summary, the authors' findings illustrate metformin's mechanism of action on SHED and confirm that SHED pre-treated with metformin exhibits a strong capacity for promoting angiogenesis. This helps in promoting the application of dental pulp-derived stem cells pre-treated with metformin in regeneration engineering.
人脱落乳牙来源的干细胞（SHED）在组织工程和再生医学中具有重要作用。血管生成在组织再生中至关重要，也是再生医学的主要目标。二甲双胍作为一线抗糖尿病药物，对干细胞具有多种有价值的影响。本研究旨在探讨二甲双胍对 SHED 介导的血管生成的影响及其作用机制。
首先，通过细胞增殖、流式细胞术、成骨、成脂和成软骨诱导以及蛋白质组学分析来探索二甲双胍在 SHED 中的作用。随后，通过体外共培养条件下经二甲双胍预处理的 SHED 的迁移和管形成测定来评估趋化性和血管生成增强，并通过定量逆转录聚合酶链反应测定相对信使 RNA 表达水平。最后，使用裸鼠进行体内管形成测定，并通过抗人 CD31 抗体的免疫组织化学染色分析切片。
二甲双胍显著促进 SHED 的增殖以及成骨、成脂和成软骨分化。蛋白质组学显示，二甲双胍显著上调了 124 种涉及细胞内过程的差异丰度蛋白，包括参与细胞迁移和血管生成的各种蛋白，如 MAPK1。共培养系统表明，经二甲双胍预处理的 SHED 显著改善了人脐静脉内皮细胞的迁移和血管生成。此外，经二甲双胍预处理的 SHED 具有更强的体内促进血管生成的能力。
综上所述，作者的研究结果说明了二甲双胍对 SHED 的作用机制，并证实经二甲双胍预处理的 SHED 具有很强的促进血管生成能力。这有助于促进经二甲双胍预处理的牙髓源性干细胞在再生工程中的应用。
