人脐带间充质干细胞与内皮细胞共移植促进体内血管生成和牙髓再生。
Cotransplantation of human umbilical cord mesenchymal stem cells and endothelial cells for angiogenesis and pulp regeneration in vivo.
机构信息
The First Affiliated Hospital of Harbin Medical University, No. 23 Youzheng Street, Nangang District, Harbin 150001, China; Department of Endodontics, School of Stomatology, Harbin Medical University, No. 143 Yiman Street, Nangang District, Harbin 150001, China.
Department of Prosthodontics, The First Affiliated Hospital of Harbin Medical University, No. 143 Yiman Street, Nangang District, Harbin 150001, China.
出版信息
Life Sci. 2020 Aug 15;255:117763. doi: 10.1016/j.lfs.2020.117763. Epub 2020 May 8.
AIMS
To explored the potential of human umbilical cord mesenchymal stem cells (hUCMSCs) as seed cells for dental pulp regeneration and the possibility of cotransplantation hUCMSCs and endothelial cells (ECs) for angiogenesis and pulp regeneration in vivo.
MATERIALS AND METHODS
hUCMSCs and human umbilical vein endothelial cells (HUVECs) were cocultured for matrigel angiogenesis assay in vitro and Matrigel plug assay in vivo. Next, we used the transwell coculture system to coculture hUCMSCs and HUVECs in vitro for RNA- sequencing (RNA-seq). Last, encapsulated hUCMSCs and HUVECs in scaffolds were injected into the root segments, and transplanted into immunodeficient mice for dental pulp regeneration.
KEY FINDINGS
In vitro Matrigel angiogenesis assay and in vivo Matrigel plug assay indicated that cocultured hUCMSCs and HUVECs promote vascular formation of HUVECs, especially in 1:5 (hUCMSCs:HUVECs) coculture group. The RNA-seq result indicated that cocultured HUVECs exhibited high Hif-1 signaling pathway activity. We performed the cell transfection assay to knock down HIF1A-AS2 in HUVECs and then coculture with hUCMSCs, and the expression of VEGFA, HIF1A and PECAM1 were reduced. In pulp regeneration assay, Cotransplantation of hUCMSCs and HUVECs (1,5) group showed pulp-like tissue regeneration.
SIGNIFICANCE
Cocultured hUCMSCs and HUVECs can promote vascular formation of HUVECs, and the optimal coculture ration is 1:5 (hUCMSCs:HUVECs). hUCMSCs promote angiogenesis of HUVECs through the long noncoding RNA HIF1A-AS2-activation of the Hif-1 signaling pathway. Cotransplantation of hUCMSCs and HUVECs can regenerate dental pulp-like tissue in vivo.
目的
探索人脐带间充质干细胞（hUCMSCs）作为牙髓再生种子细胞的潜力，以及共培养 hUCMSCs 和内皮细胞（ECs）促进体内血管生成和牙髓再生的可能性。
材料和方法
体外进行 hUCMSCs 和人脐静脉内皮细胞（HUVECs）共培养 Matrigel 血管生成实验和 Matrigel plugs 体内实验。然后，我们使用 Transwell 共培养系统体外共培养 hUCMSCs 和 HUVECs 进行 RNA 测序（RNA-seq）。最后，将包裹在支架中的 hUCMSCs 和 HUVECs 注射到牙根段，并移植到免疫缺陷小鼠体内进行牙髓再生。
主要发现
体外 Matrigel 血管生成实验和体内 Matrigel plugs 实验表明，共培养的 hUCMSCs 和 HUVECs 促进 HUVECs 的血管形成，特别是在 1:5（hUCMSCs:HUVECs）共培养组。RNA-seq 结果表明，共培养的 HUVECs 表现出高的 Hif-1 信号通路活性。我们进行了细胞转染实验，敲低 HUVECs 中的 HIF1A-AS2，然后与 hUCMSCs 共培养，结果发现 VEGFA、HIF1A 和 PECAM1 的表达减少。在牙髓再生实验中，hUCMSCs 和 HUVECs（1:5）共移植组显示出牙髓样组织再生。
意义
共培养的 hUCMSCs 和 HUVECs 可以促进 HUVECs 的血管形成，最佳共培养比例为 1:5（hUCMSCs:HUVECs）。hUCMSCs 通过长链非编码 RNA HIF1A-AS2 激活 Hif-1 信号通路促进 HUVECs 的血管生成。hUCMSCs 和 HUVECs 的共移植可以在体内再生牙髓样组织。
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