Bangkok Hospital Dental Center, Bangkok Hospital, Bangkok 10310, Thailand.
BDMS Wellness Clinic, Bangkok Dusit Medical Services, Public Company Limited, Bangkok 10330, Thailand.
Molecules. 2021 Nov 9;26(22):6756. doi: 10.3390/molecules26226756.
Dentin matrix protein 1 (DMP1) contains a large number of acidic domains, multiple phosphorylation sites, a functional arginine-glycine-aspartate (RGD) motif, and a DNA binding domain, and has been shown to play essential regulatory function in dentin and bone mineralization. DMP1 could also orchestrate bone matrix formation, but the ability of DMP1 on Ti to human mesenchymal stem cell (hMSC) conversion to osteoblasts has not been studied. There is importance to test if the DMP1 coated Ti surface would promote cell migration and attachment to the metal surface and promote the differentiation of the attached stem cells to an osteogenic lineage. This study aimed to study the human mesenchymal stem cells (hMSCs) attachment and proliferation on DMP1 coated titanium (Ti) disks compared to non-coated disks, and to assess possible osteoblastic differentiation of attached hMSCs. Sixty-eight Ti disks were divided into two groups. Group 1 disks were coated with dentin matrix protein 1 and group 2 disks served as control. Assessment with light microscopy was used to verify hMSC attachment and proliferation. Cell viability was confirmed through fluorescence microscopy and mitochondrial dehydrogenase activity. Real-time polymerase chain reaction analysis was done to study the gene expression. The proliferation assay showed significantly greater cell proliferation with DMP1 coated disks compared to the control group (-value < 0.001). Cell vitality analysis showed a greater density of live cells on DMP1 coated disks compared to the control group. Alkaline phosphatase staining revealed higher enzyme activity on DMP1 coated disks and showed itself to be significantly higher than the control group (-value < 0.001). von Kossa staining revealed higher positive areas for mineralized deposits on DMP1 coated disks than the control group (-value < 0.05). Gene expression analysis confirmed upregulation of runt-related transcription factor 2, osteoprotegerin, osteocalcin, osteopontin, and alkaline phosphatase on DMP1 coated disks (-value < 0.001). The dentin matrix protein promoted the adhesion, proliferation, facilitation differentiation of hMSC, and mineralized matrix formation.
牙本质基质蛋白 1(DMP1)含有大量酸性结构域、多个磷酸化位点、具有功能性精氨酸-甘氨酸-天冬氨酸(RGD)基序和 DNA 结合结构域,在牙本质和骨矿化中发挥重要的调节功能。DMP1 还可以协调骨基质的形成,但 DMP1 对钛表面向人间充质干细胞(hMSC)向成骨细胞转化的能力尚未得到研究。测试 DMP1 涂覆的 Ti 表面是否会促进细胞迁移和附着在金属表面上,并促进附着的干细胞向成骨谱系分化是很重要的。本研究旨在研究 DMP1 涂覆的钛(Ti)盘上的人骨髓间充质干细胞(hMSC)的附着和增殖与非涂覆盘相比,以及评估附着的 hMSC 可能的成骨细胞分化。将 68 个 Ti 盘分为两组。第 1 组盘涂有牙本质基质蛋白 1,第 2 组盘作为对照。通过光镜评估来验证 hMSC 的附着和增殖。通过荧光显微镜和线粒体脱氢酶活性确认细胞活力。通过实时聚合酶链反应分析研究基因表达。增殖试验显示,DMP1 涂覆的磁盘与对照组相比,细胞增殖显著增加(-值<0.001)。细胞活力分析显示,DMP1 涂覆的磁盘上活细胞的密度高于对照组。碱性磷酸酶染色显示,DMP1 涂覆的磁盘上的酶活性较高,明显高于对照组(-值<0.001)。茜素红 S 染色显示,DMP1 涂覆的磁盘上的矿化沉积物的阳性区域高于对照组(-值<0.05)。基因表达分析证实,DMP1 涂覆的磁盘上 runt 相关转录因子 2、骨保护素、骨钙素、骨桥蛋白和碱性磷酸酶的表达上调(-值<0.001)。牙本质基质蛋白促进 hMSC 的黏附、增殖、促进分化和矿化基质形成。