Co-cultured spheroids of human periodontal ligament mesenchymal stem cells and vascular endothelial cells enhance periodontal tissue regeneration
Co-cultured spheroids of human periodontal ligament mesenchymal stem cells and vascular endothelial cells enhance periodontal tissue regeneration
复制标题
DOI：
10.1016/j.reth.2019.12.008
复制
发表时间：
2020-06-01
影响因子：
4.3
通讯作者：
Nakashima, Keisuke
中科院分区：
文献类型：
--
作者：
Sano, Kotaro;Usui, Michihiko;Nakashima, Keisuke
前言：人牙周膜间充质干细胞(HPDLMSCs)在牙周组织的动态平衡和再生中发挥着重要作用。此外，球形细胞优于单层培养的细胞。我们在体外和体内研究了hPDLMSCs和人脐静脉内皮细胞(HUVECs)共培养球体中牙周组织再生的特点和潜力。方法：用微孔芯片制备细胞比例分别为1：1、1：2、2：1的hPDLMSCs/HUVECs共培养球体。采用实时定量聚合酶链式反应(PCR)、酶联免疫吸附试验(ELISA)和结节形成试验检测共培养的球体的性质。制备大鼠上颌骨第一磨牙牙周组织缺损区，并进行移植实验。结果：与单层和球体培养的hPDLMSCs相比，共培养的hPDLMSCs中的干细胞标志、血管内皮生长因子(VEGF)和成骨相关基因的表达水平上调。与单层和球形培养的hPDLMSCs相比，共培养的球形细胞结节形成也增加。共培养的球体在移植后4周或8周促进了新牙骨质的形成，尽管共培养的球体与hPDLMSC球体在新骨形成方面没有明显差异。结论：共培养的球体可促进牙周组织再生。HPDLMSCs与HUVECs共培养的球体可能是一种有效的诱导牙周组织再生的治疗方法。(C)2020年，日本再生医学学会。由爱思唯尔B.V.制作和托管。这是CC BY-NC-ND许可证(http://creativecommons.org/licenses/by-nc-nd/4.0中的一篇开放获取文章。
Introduction: Human periodontal ligament mesenchymal stem cells (hPDLMSCs) have been known that they play important roles in homeostasis and regeneration of periodontal tissues. Additionally, spheroids are superior to monolayer-cultured cells. We investigated the characteristics and potential of periodontal tissue regeneration in co-cultured spheroids of hPDLMSCs and human umbilical vein endothelial cells (HUVECs) in vitro and in vivo . Methods: Co-cultured spheroids were prepared with cell ratios of hPDLMSCs: HUVECs = 1:1, 1:2, and 2:1, using microwell chips. Real-time polymerase chain reaction (PCR) analysis, Enzyme-Linked Immuno Sorbent Assay (ELISA), and nodule formation assay were performed to examine the properties of co- cultured spheroids. Periodontal tissue defects were prepared in the maxillary first molars of rats and subjected to transplantation assay. Results: The expression levels of stemness markers, vascular endothelial growth factor ( VEGF ), osteogenesis-related genes were up-regulated in co-cultured spheroids, compared with monolayer and spheroid-cultured hPDLMSCs. The nodule formation was also increased in co-cultured spheroids, compared with monolayer and spheroid cultures of hPDLMSCs. Treatment with co-cultured spheroids enhanced new cementum formation after 4 or 8 weeks of transplantation, although there was no sig- ni ficant difference in the new bone formation between co-cultured spheroids and hPDLMSC spheroids. Conclusions: We found that co-cultured spheroids enhance the periodontal tissue regeneration. Co- cultured spheroids of hPDLMSCs and HUVECs may be a useful therapy that can induce periodontal tis- sue regeneration. (C) 2020, The Japanese Society for Regenerative Medicine. Production and hosting by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/ 4.0/).