Okada Hitoe, Takahashi Kosuke, Ogura Naomi, Tomoki Risa, Ito Ko, Kondoh Toshirou
Department of Maxillofacial Surgery, Nihon University School of Dentistry at Matsudo, Matsudo, Japan.
Research Institute of Oral Science, Nihon University School of Dentistry at Matsudo, Matsudo, Japan.
J Dent Sci. 2016 Sep;11(3):245-252. doi: 10.1016/j.jds.2015.12.001. Epub 2016 May 26.
BACKGROUND/PURPOSE: Plasma rich in growth factors (PRGFs), which is prepared from autologous blood from patients, has been reported with regards to bone regeneration for dental implants. Human dental follicle cells (hDFCs) have the capacity to commit to multiple cell types such as the osteoblastic lineage. The aim of this study is to evaluate the effects of PRGFs for mineralization in hDFCs.
PRGFs was prepared from whole blood centrifuged at 460 for 8 minutes. hDFCs isolated from the dental follicle with collagenase/dispase were cultured with growth medium or osteogenic induction medium (OIM) containing PRGFs or fetal bovine serum. Concentrations of the growth factors were examined using an enzyme-linked immunosorbent assay kit. A cell migration assay was used for two-dimensional movement. Gene expressions were examined with real-time polymerase chain reaction using a DyNAmo SYBR Green quantitative polymerase chain reaction kit.
The platelet concentration in PRGF Fraction 2 was 2.14-fold higher than in whole blood. White blood cells were not detected in PRGFs. Transforming growth factor-β levels were higher than insulin-like growth factor-1, platelet-derived growth factor-AB and -BB, and vascular endothelial growth factors in PRGF Fraction 2. Proliferation and migration by hDFCs increased in OIM supplemented with PRGFs in a dose-dependent manner and were higher in hDFCs cultured in OIM plus 10% PRGFs compared with OIM plus 10% fetal bovine serum. PRGFs upregulated the gene expression of , , , , and in hDFCs.
PRGFs may promote bone regeneration due to it including high levels of growth factors.
背景/目的:富含生长因子的血浆(PRGFs)由患者自体血液制备而成,已有关于其用于牙种植体骨再生的报道。人牙囊细胞(hDFCs)有分化为多种细胞类型(如成骨细胞系)的能力。本研究旨在评估PRGFs对hDFCs矿化的影响。
通过在460离心8分钟从全血中制备PRGFs。用胶原酶/ dispase从牙囊中分离出的hDFCs,在含有PRGFs或胎牛血清的生长培养基或成骨诱导培养基(OIM)中培养。使用酶联免疫吸附测定试剂盒检测生长因子的浓度。细胞迁移试验用于二维运动。使用DyNAmo SYBR Green定量聚合酶链反应试剂盒通过实时聚合酶链反应检测基因表达。
PRGF组分2中的血小板浓度比全血高2.14倍。PRGFs中未检测到白细胞。PRGF组分2中转化生长因子-β水平高于胰岛素样生长因子-1、血小板衍生生长因子-AB和-BB以及血管内皮生长因子。在补充有PRGFs的OIM中,hDFCs的增殖和迁移呈剂量依赖性增加,并且与OIM加10%胎牛血清相比,在OIM加10% PRGFs中培养的hDFCs中更高。PRGFs上调了hDFCs中、、、和的基因表达。
PRGFs可能因其含有高水平生长因子而促进骨再生。