摘要
背景:牙周膜干细胞是牙周组织中的成体干细胞,具有高度增殖、自我更新能力和多分化潜能。促进牙周膜干细胞向成骨细胞分化有助于牙周疾病的治疗。目的:观察胰岛素样生长因子1和成纤维细胞生长因子2对牙周膜干细胞向成骨细胞分化的影响。方法:采用胶原酶消化人牙周膜组织,获得牙周膜干细胞,经体外鉴定、扩增后,通过倒置显微镜、苏木精-伊红染色、流式细胞仪对牙周膜干细胞进行生物学检测。分别在成骨细胞诱导培养液中加入成骨诱导液(对照组)及胰岛素样生长因子1和成纤维细胞生长因子2持续诱导7,14d后,进行碱性磷酸酶染色、碱性磷酸酶活性检测,以及茜素红染色,并用实时定量PCR法检测向成骨细胞分化的标志性基因的表达情况。结果与结论:胰岛素样生长因子1刺激组的碱性磷酸酶活性以及钙化结节明显高于对照组,Runx2、Alp、col-1的mRNA呈高表达;成纤维细胞生长因子2刺激组的碱性磷酸酶活性以及钙化结节也高于对照组,Runx2、Alp、col-1的mRNA表达量也高于对照组。提示胰岛素样生长因子1和成纤维细胞生长因子2在不同程度上促进体外培养的牙周膜干细胞向成骨细胞方向分化。
BACKGROUND： Periodontal ligament stem cells are the adult stem cells in periodontal tissues, which possess the ability for strong proliferation, self-renewal and multipotent differentiation. These cells can promote the osteoblast differentiation of periodontal ligament stem cells and therefore assist in treatment of periodontal diseases.OBJECTIVE： To investigate the effects of insulin-like growth factor-1 and fibroblast growth factor-2 on osteoblast differentiation of periodontal ligament stem cells. METHODS： Human periodontal ligament stem cells were harvested from periodontal ligament tissue by collagenase digestion. After in vitro identification, amplification and hematoxylin-eosin staining, periodontal ligament stem cells were biologically detected by flow cytometry. Then these cells were induced with osteogenic induction medium, insulin-like growth factor-1 or fibroblast growth factor-2 for 7 and 14 days. Alkaline phosphatase staining, alkaline phosphatase activity detection and alizarin red staining were performed. In addition, real-time quantitative PCR method was performed to detect the expression of marker gene of osteogenic differentiation. RESULTS AND CONCLUSION： Compared to the control group, alkaline phosphatase activity and calcified nodules were significantly greater, and Runx2, Alp, and col-1 mRNA expression was significantly higher, in the insulin-like growth factor-1 group. These findings suggest that insulin-like growth factor-1 and fibroblast growth factor-2, to different extents, promote osteogenic differentiation of periodontal ligament stem cells cultured in vitro.
出处
《中国组织工程研究》
CSCD
2013年第10期1821-1826,共6页
Chinese Journal of Tissue Engineering Research
关键词
干细胞
干细胞培养与分化
胰岛素样生长因子1
成纤维细胞生长因子2
牙周膜干细胞
成骨细胞
成骨分化
软骨
骨生长
多分化
钙化结节
干细胞图片文章
stem cells
stem cell culture and differentiation
insulin-like growth factor-1
fibroblast growth factor-2
periodontal ligament stem cells
osteoblasts
osteogenic differentiation
cartilage
bone growth
multipotent differentiation
calcified nodules
stem cell photographs-containing paper
