Division of Biochemistry, Department of Oral Biology, School of Dentistry, Health Sciences University of Hokkaido, Hokkaido, Japan.
Division of Clinical Cariology and Endodontology, Department of Oral Rehabilitation, School of Dentistry, Health Sciences University of Hokkaido, Hokkaido, Japan.
J Endod. 2018 Sep;44(9):1367-1375. doi: 10.1016/j.joen.2018.05.018.
iMatrix-511 is a novel integrin-binding fragment derived from laminin-511. Previous studies showed its superiority as a culture substrate for xeno-free culture and maintenance of pluripotency in stem cells. However, its effects in the dental field remain largely unknown. The aim of the present study was to unravel the in vitro effects of iMatrix-511 in comparison with vitronectin (VN).
Biochemical assays were performed in vitro in MDPC-23 cells. The optimal coating density for 2 proteins was determined using the cell counting kit-8. To evaluate cell proliferation to both proteins, MDPC-23 cells were directly seeded onto the iMatrix-511 or VN-modified polystyrene and analyzed by the cell counting kit-8. The phenotype of cells seeded on iMatrix-511 and VN was characterized. Phenotypic characterization included real-time reverse-transcription polymerase chain reaction and alizarin red staining.
The optimal coating density for iMatrix-511 and VN was determined to be 1 μg/cm and 0.25 μg/cm, respectively. Cells cultured on iMatrix-511 showed higher cell proliferative activity than the noncoated control and VN on days 1, 2, and 4. Cell morphology observation revealed MDPC-23 cells attach preferentially to iMatrix-511 and start to spread as early as 1 hour after inoculation. MDPC-23 cells exhibited more potent odontogenic differentiation on iMatrix-511 than the control and VN as shown by the marked enhancement of dentin matrix protein 1 and dentin sialophosphoprotein messenger RNA expression. Although both proteins showed more mineralized nodule formation than the control, iMatrix-511 remained to be the one that elicited stronger calcific deposition.
iMatrix-511 supported the proliferation and acquisition of odontogenic cell phenotype in vitro, rendering this novel material a potential candidate for dentin regeneration.
iMatrix-511 是一种新型整合素结合片段，来源于层粘连蛋白-511。先前的研究表明，它作为无动物培养基质培养干细胞的多能性具有优越性。然而，其在牙科领域的影响在很大程度上尚不清楚。本研究旨在揭示 iMatrix-511 与 vitronectin（VN）相比的体外作用。
在 MDPC-23 细胞中进行体外生化分析。使用细胞计数试剂盒-8 确定 2 种蛋白质的最佳涂层密度。为了评估两种蛋白质对细胞增殖的影响，将 MDPC-23 细胞直接接种到 iMatrix-511 或 VN 修饰的聚苯乙烯上，并通过细胞计数试剂盒-8 进行分析。对接种在 iMatrix-511 和 VN 上的细胞表型进行了特征描述。表型特征包括实时逆转录聚合酶链反应和茜素红染色。
确定 iMatrix-511 和 VN 的最佳涂层密度分别为 1μg/cm 和 0.25μg/cm。第 1、2 和 4 天，在 iMatrix-511 上培养的细胞显示出比非涂层对照和 VN 更高的细胞增殖活性。细胞形态观察显示，MDPC-23 细胞优先附着在 iMatrix-511 上，并在接种后 1 小时内开始扩散。与对照组和 VN 相比，MDPC-23 细胞在 iMatrix-511 上表现出更强的成牙本质分化能力，表现为牙本质基质蛋白 1 和牙本质涎磷蛋白信使 RNA 表达的显著增强。尽管两种蛋白质的矿化结节形成均多于对照组，但 iMatrix-511 仍然是一种能引起更强钙化沉积的物质。
iMatrix-511 支持体外细胞增殖和成牙本质细胞表型的获得，使这种新型材料成为牙本质再生的潜在候选物。
