Choi Hyunmin, Park Kyu-Hyung, Lee Ah-Reum, Mun Chin Hee, Shin Yong Dae, Park Yong-Beom, Park Young-Bum
a Department of Prosthodontics, Oral Science Research Center, BK21 Plus Project , Yonsei University College of Dentistry , Seoul , Korea.
b Division of Rheumatology, Department of Internal Medicine , Institute for Immunology and Immunological Disease, Yonsei University College of Medicine , Seoul , Korea.
Acta Odontol Scand. 2017 Jul;75(5):309-318. doi: 10.1080/00016357.2017.1303847. Epub 2017 Mar 23.
The aim of this study is to investigate the behaviour of iPSc derived from dental stem cells in terms of initial adhesion, differentiation potential on differently surface-treated titanium disc.
iPSc derived from human gingival fibroblasts (hGFs) were established using 4-reprogramming factors transduction with Sendai virus. The hGF-iPSc established in this study exhibited the morphology and growth properties similar to human embryonic stem (ES) cells and expressed pluripotency makers. Alkaline Phosphatase (AP) staining, Embryoid Body (EB) formation and in vitro differentiation and karyotyping further confirmed pluripotency of hGF-iPSc. Then, hGF-iPSc were cultured on machined- and Sandblasted and acid etched (SLA)-treated titanium discs with osteogenic induction medium and their morphological as well as quantitative changes according to different surface types were investigated using Alizrin Red S staining, Scanning electron microscopy (SEM), Flow cytometry and RT-PCR.
Time-dependent and surface-dependent morphological changes as well as quantitative change in osteogenic differentiation of hGF-iPSc were identified and osteogenic gene expression of hGF-iPSc cultured on SLA-treated titanium disc found to be greater than machined titanium disc, suggesting the fate of hGF-iPSc may be determined by the characteristics of surface to which hGF-iPSc first adhere.
iPSc derived from dental stem cell can be one of the most promising and practical cell sources for personalized regenerative dentistry and their morphological change as well as quantitative change in osteogenic differentiation according to different surface types may be further utilized for future clinical application incorporated with dental implant.
本研究旨在探讨源自牙干细胞的诱导多能干细胞（iPSc）在初始黏附方面的行为，以及在不同表面处理的钛盘上的分化潜能。
利用仙台病毒转导4种重编程因子，建立源自人牙龈成纤维细胞（hGFs）的iPSc。本研究中建立的hGF-iPSc表现出与人类胚胎干细胞（ES细胞）相似的形态和生长特性，并表达多能性标志物。碱性磷酸酶（AP）染色、胚体（EB）形成、体外分化和核型分析进一步证实了hGF-iPSc的多能性。然后，将hGF-iPSc接种在机械加工和喷砂酸蚀（SLA）处理的钛盘上，加入成骨诱导培养基，采用茜素红S染色、扫描电子显微镜（SEM）、流式细胞术和逆转录-聚合酶链反应（RT-PCR）研究其根据不同表面类型的形态和定量变化。
确定了hGF-iPSc成骨分化的时间依赖性和表面依赖性形态变化以及定量变化，发现接种在SLA处理钛盘上的hGF-iPSc的成骨基因表达高于机械加工钛盘，这表明hGF-iPSc的命运可能由其最初黏附的表面特性决定。
源自牙干细胞的iPSc可能是个性化再生牙科最有前景和实用的细胞来源之一，其根据不同表面类型的形态变化和成骨分化的定量变化可能会在未来结合牙种植体的临床应用中得到进一步利用。
