脱细胞真皮基质对年轻恒牙牙乳头干细胞分化的作用

发布时间：2018-06-17 02:11

本文选题：牙乳头 + 干细胞　；参考：《北京大学学报(医学版)》2014年01期

【摘要】：目的:获取年轻恒牙牙乳头组织,进行细胞培养以及与脱细胞真皮基质(acellular dermal matrix,ADM)的复合培养,了解ADM在干细胞成骨、成脂转化中的作用。方法:利用口腔外科手段获取年轻恒牙牙乳头组织,分离培养细胞,对其进行流式细胞分析,细胞分为两组,并通过染色进行观察,利用实时荧光定量聚合酶链反应(realtime polymerase chain reaction,real-time PCR)对特定蛋白的RNA水平进行分析。将牙乳头细胞复合ADM培养,实验组对培养细胞进行成骨与成脂诱导分化,对照组不进行诱导,同样进行染色与real-time PCR分析。结果:牙乳头细胞生长活跃,利用流式细胞分析发现70.3%细胞的间充质干细胞表面标志物Stro-1表达呈阳性,有96%细胞CD105表达呈阳性,经过诱导后,实验组细胞与成骨相关的骨钙素(osteocalcin,OCN)和骨涎蛋白(bone sialoprotein,BSP)的RNA水平,与成脂相关的肝X-受体α(liver X-recepterα,LXRα)、脂蛋白脂酶(lipoprotein lipase,LPL)、过氧化物酶体增殖物激活受体γ(peroxisome proliferator activated receptorγ,PPAR-γ)和B类清道夫受体1(scavenger receptor class B type 1,SR-B1)的RNA水平均高于未经诱导的对照组细胞。牙乳头细胞复合ADM培养后细胞仍然生长活跃,两者紧密贴附,经诱导分化,实验组OCN与BSP的RNA水平高于对照组(P0.05),实验组LPL的RNA水平高于对照组(P0.05)。结论:通过口腔外科获得的年轻恒牙牙乳头组织含有大量具有成骨能力的干细胞,将其与具有胶原支架结构的ADM复合培养,发现ADM可以作为干细胞生长的支架结构,并对干细胞的成骨诱导起到一定作用。
[Abstract]:Objective: to investigate the role of dermal in osteogenesis and adipogenic transformation of adult permanent teeth by cell culture and co-culture with acellular dermal matrix adm. Methods: the papilla tissue of young permanent teeth was obtained by oral surgery and cultured cells were isolated and analyzed by flow cytometry. The cells were divided into two groups and observed by staining. The RNA level of specific protein was analyzed by real time polymerase chain real-time PCR. Dental papilla cells were cultured with ADM. The cultured cells were induced by osteogenesis and adipogenic differentiation in the experimental group, but not in the control group. The same staining and real-time analysis were carried out in the control group. Results: dental papilla cells were active in growth. The expression of Stro-1 on mesenchymal stem cell surface was positive in 70.3% of the cells, and CD105 was positive in 96% of the cells. The RNA levels of osteocalcin (OCN) and sialoprotein (BSPP) of osteoblast-associated osteocalcin in experimental group were observed. The levels of LXR 伪, lipoprotein lipase, peroxisome proliferator activated receptor 纬 -PPAR- 纬 and 1(scavenger receptor class B type 1 SR-B1) were significantly higher in lipopolysaccharide (LXR), lipoprotein lipase, peroxisome proliferator activated receptor 纬 (PPAR- 纬) and class B scavenger receptor (1(scavenger receptor class B type 1r-B1) than in uninduced control cells. After the dental papilla cells were cultured with ADM, the cells were still active. The mRNA levels of OCN and BSP in the experimental group were higher than those in the control group (P 0.05), and the levels of LPL RNA in the experimental group were higher than those in the control group (P 0.05). Conclusion: the dental papilla of young permanent teeth obtained by oral surgery contains a large number of osteogenic stem cells. It is found that ADM can be used as a scaffold for stem cell growth by co-culture with ADM with collagen scaffold. It also plays a certain role in osteogenesis induction of stem cells.
【作者单位】：北京大学口腔医学院·口腔医院口腔颌面外科;
【分类号】：R781

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