Nugraha Alexander Patera, Narmada Ida Bagus, Ernawati Diah Savitri, Dinaryanti Aristika, Hendrianto Eryk, Ihsan Igo Syaiful, Riawan Wibi, Rantam Fedik Abdul
Graduate School of Immunology, Postgraduate School, Universitas Airlangga, Surabaya, 60132, Indonesia.
Orthodontic Department, Faculty of Dental Medicine, Universitas Airlangga, Surabaya, 60132, Indonesia.
F1000Res. 2018 Jul 25;7:1134. doi: 10.12688/f1000research.15423.1. eCollection 2018.
Alveolar bone defect regeneration has long been problematic in the field of dentistry. Gingival stromal progenitor cells (GSPCs) offer a promising solution for alveolar bone regeneration. In order to optimally differentiate and proliferate progenitor cells, growth factors (GFs) are required. Platelet rich fibrin (PRF) has many GFs and can be easily manufactured. Core-binding factor subunit-α1 (CBF-α1) constitutes a well-known osteogenic differentiation transcription factor in SPCs. Sox9, as a chondrogenic transcription factor, interacts and inhibits CBF-α1, but its precise role in direct osteogenesis remains unknown. GSPCs cultured in PRF to optimally stimulate osteogenic differentiation has been largely overlooked. The aim of this study was to analyze GSPCs cultured in PRF osteogenic differentiation predicted by CBF-α1/Sox9. : This study used a true experimental with post-test only control group design and random sampling. GPSCs isolated from the lower gingiva of four healthy, 250-gram, 1-month old, male Wistar rats ( ) were cultured for two weeks, passaged every 4-5 days. GSPCs in passage 3-5 were cultured in five M24 plates (N=108; n=6/group) for Day 7, Day 14, and Day 21 in three different mediums (control negative group: αModified Eagle Medium; control positive group: High Glucose-Dulbecco's Modified Eagle Medium (DMEM-HG) + osteogenic medium; Treatment group: DMEM-HG + osteogenic medium + PRF). CBF-α1 and Sox9 were examined with ICC monoclonal antibody. A one-way ANOVA continued with Tukey HSD test (p<0.05) based on Kolmogorov-Smirnov and Levene's tests (p>0.05) was performed. The treatment group showed the highest CBF-α1/Sox9 ratio (16.00±3.000/14.33±2.517) on Day 7, while the lowest CBF-α1/Sox9 ratio (3.33±1.528/3.67±1.155) occurred in the control negative group on Day 21, with significant difference between the groups (p<0.05). GSPCs cultured in PRF had potential osteogenic differentiation ability predicted by the CBF-α1/sox9 ratio.
牙槽骨缺损修复长期以来一直是牙科学领域的难题。牙龈基质祖细胞(GSPCs)为牙槽骨再生提供了一个有前景的解决方案。为了使祖细胞实现最佳分化和增殖,需要生长因子(GFs)。富血小板纤维蛋白(PRF)含有多种生长因子且易于制备。核心结合因子亚基-α1(CBF-α1)是众所周知的间充质干细胞(SPCs)成骨分化转录因子。Sox9作为一种软骨形成转录因子,与CBF-α1相互作用并抑制其活性,但其在直接成骨过程中的具体作用尚不清楚。在PRF中培养GSPCs以最佳刺激其成骨分化这一点在很大程度上被忽视了。本研究的目的是分析在PRF中培养的GSPCs的成骨分化情况,并通过CBF-α1/Sox9进行预测。:本研究采用仅设后测对照组的真实验设计和随机抽样。从4只健康的、体重250克、1月龄雄性Wistar大鼠的下牙龈分离出牙龈基质祖细胞(GPSCs),培养两周,每4 - 5天传代一次。将第3 - 5代的GSPCs接种于5个M24培养板中(N = 108;每组n = 6),分别在第7天、第14天和第21天置于三种不同培养基中培养(对照阴性组:α改良伊格尔培养基;对照阳性组:高糖 - 杜尔贝科改良伊格尔培养基(DMEM - HG)+成骨培养基;处理组:DMEM - HG +成骨培养基+ PRF)。采用免疫细胞化学(ICC)单克隆抗体检测CBF-α1和Sox9。基于柯尔莫哥洛夫 - 斯米尔诺夫检验和莱文检验(p > 0.05),进行单因素方差分析,并继续进行图基 Honestly significant difference(HSD)检验(p < 0.05)。处理组在第7天显示出最高的CBF-α1/Sox9比值(16.00±3.000/14.33±2.517),而对照阴性组在第21天出现最低的CBF-α1/Sox9比值(3.33±1.528/3.67±1.155),组间差异显著(p < 0.05)。在PRF中培养的GSPCs具有通过CBF-α1/sox9比值预测的潜在成骨分化能力。