人牙周膜细胞微球体在壳聚糖和聚乙烯醇膜上培养的生物学特性。

Biological properties of human periodontal ligament cell spheroids cultivated on chitosan and polyvinyl alcohol membranes.

机构信息

Department of Biomedical Engineering, National Taiwan University, Taipei, Taiwan.

School of Dentistry, National Taiwan University, Taipei, Taiwan; Department of Dentistry, National Taiwan University Hospital, Taipei, Taiwan.

出版信息

J Formos Med Assoc. 2022 Nov;121(11):2191-2202. doi: 10.1016/j.jfma.2022.03.014. Epub 2022 Apr 2.

DOI:10.1016/j.jfma.2022.03.014

PMID:35382964

Abstract

BACKGROUND/PURPOSE: Multicellular spheroid cultures have attracted increasing attention in the field of periodontal regeneration. However, very few studies have reported the periodontal ligament (PDL) cell spheroid formation via biomaterials-induced processes. This study investigated the biological characteristics of human PDL cell spheroids formed on two hydrophilic polymer-based biomaterials, namely chitosan and polyvinyl alcohol.

METHODS

The expressions of periostin, paxillin, hypoxia-inducible factor 1-α (HIF-1α), and vascular endothelial growth factor (VEGF) were analyzed. Cell migration ability was assessed using a scratch assay. Furthermore, PDL cell spheroids were cultured in 3D-printed polylactic acid scaffolds to evaluate mineralizing capability.

RESULTS

Western blot analysis revealed increased expressions of periostin, HIF-1α, and VEGF in the 3D spheroids. After the spheroids were reseeded, the cells gradually migrated outward from the spheroids and time-dependent distribution of paxillin was observed. The cells migrating outward from the 3D spheroids demonstrated greater migration ability than that of 2D monolayer cells. Compared to the dissociated cells from a monolayer culture, the cell spheroids formed on the chitosan membrane exhibited elevated alkaline phosphatase activity and an increase in mineralized matrix deposition.

CONCLUSION

The biomaterial-induced formation of PDL cell spheroids suggests a novel strategy for cell delivery in research and clinical applications of periodontal regeneration.

摘要

背景/目的：在牙周再生领域，多细胞球体培养受到了越来越多的关注。然而，很少有研究报道过通过生物材料诱导过程形成牙周韧带（PDL）细胞球体。本研究调查了在两种亲水聚合物基生物材料（壳聚糖和聚乙烯醇）上形成的人 PDL 细胞球体的生物学特性。

方法

分析了骨桥蛋白、桩蛋白、缺氧诱导因子 1-α（HIF-1α）和血管内皮生长因子（VEGF）的表达。通过划痕实验评估细胞迁移能力。此外，将 PDL 细胞球体培养在 3D 打印的聚乳酸支架上，以评估矿化能力。

结果

Western blot 分析显示，3D 球体中骨桥蛋白、HIF-1α 和 VEGF 的表达增加。球体重新接种后，细胞逐渐从球体向外迁移，并观察到桩蛋白的时间依赖性分布。从 3D 球体中迁移出来的细胞比 2D 单层细胞具有更强的迁移能力。与单层培养物中分离的细胞相比，壳聚糖膜上形成的细胞球体表现出更高的碱性磷酸酶活性和矿化基质沉积增加。