机械牵张应力对人根尖牙乳头干细胞增殖分化的影响

发布时间：2018-04-02 15:17

本文选题：机械牵张应力　切入点：根尖牙乳头干细胞　出处：《医学研究生学报》2017年10期

【摘要】：目的根尖牙乳头干细胞(SCAP)被看作是根尖周组织再生的种子细胞并被用于以干细胞为基础的根尖周组织再生工程中。文章探讨不同力值的机械牵张应力对人SCAP增殖、分化潜能的影响。方法利用组织块酶消化法及有限稀释法分离、培养人SCAP并加以鉴定。根据对细胞加载机械牵张应力大小不同将实验分为150、200、250 g组,另设空白对照组(未做加力处理)。利用MTT法检测不同大小静态机械牵张应力刺激对SCAP增殖活性的影响。利用Western blot检测机械牵张应力作用下SCAP成骨/成牙分化相关蛋白(ALP、OSX、DSP)表达的变化以及不同大小静态机械牵张应力作用下SCAP内质网应激分子伴侣GRP78的表达情况。结果 SCAP细胞经过3周的成骨诱导后进行茜素红染色,镜下可见块状矿化结节的出现,SCAP细胞经过2周的成脂诱导后进行油红O染色,镜下可见红染的脂滴出现。SCAP细胞表面抗原表型通过流式细胞仪检测分析显示:SCAP对CD31(2.46%,内皮细胞标记物)和CD45(0.07%,造血干细胞标记物)呈阴性表达,对CD90(99.89%,间质细胞标记)和STRO-1(15.21%,干细胞标记)呈阳性表达。与150 g组比较,200 g加力组的加载机械牵张应力第2天,SCAP增殖能力明显下降(P0.05);250 g加力组的加载机械牵张应力的第2、3、4、5天,SCAP增殖能力显著下降(P0.05)。与200 g组比较,250 g加力组的加载机械牵张应力的第2、3、4、5天,SCAP增殖能力显著下降(P0.05)。Western blot检测结果显示:加载牵张应力的第5天,与对照组相比,150 g组、200 g组、250 g组成骨分化相关蛋白ALP和OSX和DSP表达均显著升高(P0.05)。与150 g组相比,200g组ALP、OSX表达差异无统计学意义(P0.05),DSP表达显著升高(P0.05);250 g组ALP、OSX、DSP表达均显著降低(P0.05)。与200 g组相比,250 g组ALP、OSX、DSP表达均显著降低(P0.05)。Western blot检测显示:加载牵张应力的第5天,与对照组GRP78表达(0.279±0.085)相比,150、200、250 g组GRP78表达(1.085±0.128、1.289±0.076、0.810±0.067)均显著升高(P0.05)。结论机械牵张应力对人根尖牙乳头干细胞的增殖和成骨/成牙本质分化具有调控作用。内质网应激参与了机械牵张应力作用下的SCAP成骨/成牙分化过程,并促进了SCAP成骨/成牙分化。
[Abstract]:Objective SCAPs are regarded as seed cells of periapical tissue regeneration and used in stem cell-based periapical tissue regeneration engineering.The effect of mechanical stretch stress on the proliferation and differentiation potential of human SCAP was studied.Methods Human SCAP was isolated and identified by tissue enzyme digestion and limited dilution method.The experiment was divided into 150200250 g group and blank control group according to the different mechanical tensile stress of cell loading.The effect of static mechanical stretch stress on the proliferation of SCAP was detected by MTT method.Western blot was used to detect the expression of SCAP osteoblast / odontogenic differentiation related protein (ALP) and the expression of SCAP endoplasmic reticulum stress molecular companion (GRP78) under different static mechanical stretch stress.Results after 3 weeks of osteogenesis, SCAP cells were stained with alizarin red, and the presence of massive mineralized nodules was detected by oil red O staining after 2 weeks of adipogenic induction.The positive expression of CD90 ~ (99.89) and STRO-1N 15.21 ~ (21) in interstitial cells were observed.Compared with 200g group, the proliferative ability of SCAP in 250g group was significantly decreased on the 5th day of loading mechanical tensile stress. The results of Western blot showed that the tensile stress was 5 days after loading.Compared with the control group, the expression of osteogenic differentiation related protein ALP, OSX and DSP in the 150g group and 200g group were significantly higher than those in the control group.Compared with the 150 g group, there was no significant difference in the expression of ALP OSX in the 200g group. There was no significant increase in the DSP expression of P0.05T in the 200g group. The expression of DSP in the P0.05G group was significantly lower than that in the 250g group. The expression of DSP in the P0.05G group was significantly lower than that in the P0.05G group.Conclusion Mechanical stretch stress can regulate the proliferation and osteogenesis / dentin differentiation of human root canine papillary stem cells.Endoplasmic reticulum stress was involved in the process of SCAP osteogenesis / tooth differentiation under mechanical distraction stress and promoted SCAP osteogenesis / tooth differentiation.
【作者单位】：西南医科大学附属口腔医院正畸科;
【基金】：泸州市政府-四川医科大学联合项目[2015LZCYD-S05(1/12)]
【分类号】：R783.5

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