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  本文選題：機械牽張應力　切入點：根尖牙乳頭干細胞　出處：《醫(yī)學研究生學報》2017年10期

【摘要】：目的根尖牙乳頭干細胞(SCAP)被看作是根尖周組織再生的種子細胞并被用于以干細胞為基礎的根尖周組織再生工程中。文章探討不同力值的機械牽張應力對人SCAP增殖、分化潛能的影響。方法利用組織塊酶消化法及有限稀釋法分離、培養(yǎng)人SCAP并加以鑒定。根據(jù)對細胞加載機械牽張應力大小不同將實驗分為150、200、250 g組,另設空白對照組(未做加力處理)。利用MTT法檢測不同大小靜態(tài)機械牽張應力刺激對SCAP增殖活性的影響。利用Western blot檢測機械牽張應力作用下SCAP成骨/成牙分化相關蛋白(ALP、OSX、DSP)表達的變化以及不同大小靜態(tài)機械牽張應力作用下SCAP內(nèi)質(zhì)網(wǎng)應激分子伴侶GRP78的表達情況。結(jié)果 SCAP細胞經(jīng)過3周的成骨誘導后進行茜素紅染色,鏡下可見塊狀礦化結(jié)節(jié)的出現(xiàn),SCAP細胞經(jīng)過2周的成脂誘導后進行油紅O染色,鏡下可見紅染的脂滴出現(xiàn)。SCAP細胞表面抗原表型通過流式細胞儀檢測分析顯示:SCAP對CD31(2.46%,內(nèi)皮細胞標記物)和CD45(0.07%,造血干細胞標記物)呈陰性表達,對CD90(99.89%,間質(zhì)細胞標記)和STRO-1(15.21%,干細胞標記)呈陽性表達。與150 g組比較,200 g加力組的加載機械牽張應力第2天,SCAP增殖能力明顯下降(P0.05);250 g加力組的加載機械牽張應力的第2、3、4、5天,SCAP增殖能力顯著下降(P0.05)。與200 g組比較,250 g加力組的加載機械牽張應力的第2、3、4、5天,SCAP增殖能力顯著下降(P0.05)。Western blot檢測結(jié)果顯示:加載牽張應力的第5天,與對照組相比,150 g組、200 g組、250 g組成骨分化相關蛋白ALP和OSX和DSP表達均顯著升高(P0.05)。與150 g組相比,200g組ALP、OSX表達差異無統(tǒng)計學意義(P0.05),DSP表達顯著升高(P0.05);250 g組ALP、OSX、DSP表達均顯著降低(P0.05)。與200 g組相比,250 g組ALP、OSX、DSP表達均顯著降低(P0.05)。Western blot檢測顯示:加載牽張應力的第5天,與對照組GRP78表達(0.279±0.085)相比,150、200、250 g組GRP78表達(1.085±0.128、1.289±0.076、0.810±0.067)均顯著升高(P0.05)。結(jié)論機械牽張應力對人根尖牙乳頭干細胞的增殖和成骨/成牙本質(zhì)分化具有調(diào)控作用。內(nèi)質(zhì)網(wǎng)應激參與了機械牽張應力作用下的SCAP成骨/成牙分化過程,并促進了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.
【作者單位】： 西南醫(yī)科大學附屬口腔醫(yī)院正畸科;
【基金】：瀘州市政府-四川醫(yī)科大學聯(lián)合項目[2015LZCYD-S05(1/12)]
【分類號】：R783.5

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本文編號：1700971