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论文摘要

壳聚糖季铵盐对人牙周膜成纤维细胞增殖及细胞周期影响的实验研究

Effects of quaternized chitosan on the cell proliferation and cell cycle of human periodontal ligament cells

作者:吉秋霞 袁昌青 宋文斌 武宏

Author:Ji Qiuxia, Yuan Changqing, Song Wenbin, Wu Hong.

收稿日期:2012-12-12          年卷(期)页码:2013,40(5):588-591

期刊名称:国际口腔医学杂志

Journal Name:International Journal of Stomatology

关键字:壳聚糖,牙周膜成纤维细胞,细胞增殖,细胞周期,

Key words:chitosan,human periodontal ligment cell,cell proliferation,cell cycle,

基金项目:

山东省优秀中青年科学家奖励基金资助项目(BS2010CL015)

中文摘要

目的 观察不同浓度壳聚糖季铵盐(HTCC)对人牙周膜成纤维细胞(HPDLC)增殖和细胞周期的影响,并与壳聚糖(CS)相比较,为HTCC的合理应用提供依据。方法 原代培养人牙周膜成纤维细胞,免疫组织化学鉴定细胞来源。四甲基偶氮唑盐(MTT)法测定质量浓度分别为2 000、1 000、100、50、10、3、1.5、1、0.5 和0.2 mg•L-1的HTCC和CS对HPDLC 培养1、3、5 d细胞增殖活性的影响;流式细胞术测定质量浓度为3 g•L-1和 3 mg•L-1的HTCC和CS及1 g•L-1的HTCC对HPDLC细胞周期的影响。应用SSPS 13.0软件包对数据进行统计学分析。结果 经免疫组织化学显示:培养的细胞波形丝蛋白染色阳性,角蛋白染色阴性,证明细胞来源于外胚层。MTT 实验表明,与对照组相比,HTCC在质量浓度为2 000、1 000、100和50 mg•L-1时,光密度(OD)值降低;质量浓度为10、3、1.5、1、0.5 和0.2 mg•L-1时,OD值增高。而CS在所有实验质量浓度范围内均能促进细胞的增殖,在相对高质量浓度培养5 d 后对HPDLC的促增殖作用与对照组的差异具有统计学意义(P

英文摘要

Objective To evaluate the effects of quaternized chitosan[N-(2-hydroxy-3-trimethylammonium) propylchitosan chloride] (HTCC) on the proliferation and cell cycle activities of human periodontal ligament cells(HPDLCs) with different concentrations compared with chitosan(CS). Investigating the mechanisms underlying the use of HTCC is necessary. Methods Primary cells were isolated from human periodontal ligament. HPDLCs were cultured and characterized by immunocytochemistry. The effect of varying concentrations(2 000, 1 000, 100, 50, 10, 3, 1.5, 1, 0.5, and 0.2 mg•L-1) of HTCC and CS on HPDLCs were evaluated by methyl thiazolyl tetrazolium(MTT) assay. Flow cytometry (FCM) was used to determine the cell cycle states of HPDLCs with HTCC(3 g•L-1, 1 g•L-1, and 3 mg•L-1) and CS(3 g•L-1 and 3 mg•L-1). The SPSS 13.0 software package was used for statistical analysis. Results Immunocytochemistry results show that the cells stained positively to antibodies against vimentin and stained negatively to antibodies against cytokeratin, indicating that the cells were external embryo mesenchymal cells. The MTT assay showed decreased optical density(OD) values of HTCC at concentrations of 2 000, 1 000, 100, and 50 mg•L-1 and increased OD values at concentrations of 10, 3, 1.5, 1, 0.5, and 0.2 mg•L-1. CS accelerated the proliferation of HPDLC at all tested concentrations. Compared with the control, a significant difference was observed at 5 d(P

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