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 significant 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 tissue regeneration.

中文翻译：

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人牙周膜间充质干细胞和血管内皮细胞的共培养球体增强牙周组织再生。

介绍

已知人牙周膜间充质干细胞（hPDLMSCs）在牙周组织的稳态和再生中起重要作用。此外，球体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征和潜力。

方法

使用微孔芯片以 hPDLMSCs: HUVECs = 1:1、1:2 和 2:1 的细胞比例制备共培养的球体。进行实时聚合酶链反应 (PCR) 分析、酶联免疫吸附试验 (ELISA) 和结节形成试验以检查共培养球体的特性。在大鼠上颌第一磨牙中制备牙周组织缺损并进行移植试验。

结果

与单层和球体培养的 hPDLMSCs 相比，干性标志物、血管内皮生长因子 ( VEGF )、成骨相关基因的表达水平在共培养的球体中上调。与 hPDLMSCs 的单层和球体培养物相比，共培养球体中的结节形成也增加。在移植 4 或 8 周后，用共培养球体处理增强了新牙骨质的形成，尽管共培养球体和 hPDLMSC 球体之间的新骨形成没有显着差异。

结论

我们发现共培养的球体增强了牙周组织的再生。hPDLMSCs 和 HUVECs 的共培养球体可能是一种有用的治疗方法，可以诱导牙周组织再生。