發(fā)布時(shí)間：2018-04-25 12:32

  本文選題：髁突軟骨細(xì)胞 + 鼻中隔軟骨細(xì)胞　； 參考：《昆明醫(yī)科大學(xué)》2017年碩士論文

【摘要】：[目的]建立靜磁場(chǎng)作用下SD乳鼠原代下頜髁突軟骨細(xì)胞(Mandibular Condylar Chondroc-yte,MCC)和鼻中隔軟骨細(xì)胞(Nasal Septal Chondrocyte,NSC)體外培養(yǎng)模型,觀察靜磁場(chǎng)對(duì)髁突軟骨細(xì)胞和鼻中隔軟骨細(xì)胞軟骨形成的影響及差異,為進(jìn)一步深入研究靜磁場(chǎng)作用下Flrt-2對(duì)髁突軟骨和鼻中隔軟骨形成過(guò)程的影響及內(nèi)部調(diào)控機(jī)制提供指導(dǎo),為后續(xù)探討靜磁場(chǎng)對(duì)頜面部生長(zhǎng)發(fā)育的影響提供理論基礎(chǔ)。[方法]取出生1～3天的SD乳鼠髁突軟骨和鼻中隔軟骨,用機(jī)械分離和酶聯(lián)消化法獲取原代細(xì)胞,在相同培養(yǎng)條件下進(jìn)行原代培養(yǎng)和軟骨細(xì)胞鑒定。對(duì)兩種細(xì)胞分別加載280mT靜磁場(chǎng),通過(guò)阿辛藍(lán)染色檢測(cè)靜磁場(chǎng)對(duì)兩種軟骨細(xì)胞糖胺聚糖(Glycosaminoglycan,GAG)表達(dá)差異的影響;通過(guò)CCK-8(Cell Counting Kit-8,CCK-8)比色法描繪軟骨細(xì)胞生長(zhǎng)曲線,檢測(cè)靜磁場(chǎng)對(duì)兩種軟骨細(xì)胞增殖的影響差異;劃痕實(shí)驗(yàn)觀察靜磁場(chǎng)作用下兩種細(xì)胞劃痕24h后細(xì)胞遷移的能力;蛋白質(zhì)印跡法(WesternBlot,WB)檢測(cè)加載靜磁場(chǎng)后兩種軟骨細(xì)胞Flrt-2和N-cadherin蛋白的表達(dá)差異。[結(jié)果]采用相同的消化方式及培養(yǎng)條件成功培養(yǎng)出原代髁突軟骨細(xì)胞和鼻中隔軟骨細(xì)胞,Ⅱ型膠原免疫組化染色和阿辛藍(lán)染色細(xì)胞鑒定結(jié)果均呈陽(yáng)性。兩種軟骨細(xì)胞均分為空白組和磁場(chǎng)組,在大部分細(xì)胞貼壁后,磁場(chǎng)組細(xì)胞載280mT磁場(chǎng)繼續(xù)培養(yǎng)。四組軟骨細(xì)胞在第1、3、5、8天時(shí)進(jìn)行阿辛藍(lán)染色,染色結(jié)果均呈陽(yáng)性,鼻中隔軟骨細(xì)胞糖胺聚糖表達(dá)量及軟骨結(jié)節(jié)形成能力均高于髁突軟骨細(xì)胞,兩種細(xì)胞加載磁場(chǎng)組糖胺聚糖表達(dá)量及軟骨結(jié)節(jié)形成能力均較空白組細(xì)胞高;四組軟骨細(xì)胞CCK-8比色法檢測(cè)出兩種軟骨細(xì)胞的生長(zhǎng)曲線基本一致,鼻中隔軟骨細(xì)胞的增殖能力較髁突軟骨細(xì)胞強(qiáng),加載靜磁場(chǎng)后兩種軟骨細(xì)胞增殖能力均增強(qiáng);24h劃痕實(shí)驗(yàn)結(jié)果顯示鼻中隔軟骨細(xì)胞遷移能力強(qiáng)于髁突軟骨細(xì)胞,磁場(chǎng)組細(xì)胞遷移能力強(qiáng)于空白組細(xì)胞。兩種細(xì)胞在培養(yǎng)第1、3、5、8天時(shí)對(duì)Flrt-2、N-cadherin蛋白表達(dá)進(jìn)行檢測(cè),發(fā)現(xiàn)在軟骨細(xì)胞凝集期,Flrt-2蛋白表達(dá)下調(diào),N-cadherin上調(diào);凝集期過(guò)后Flrt-2蛋白表達(dá)上調(diào),N-cadherin下調(diào)。加載靜磁場(chǎng)后,兩種軟骨細(xì)胞Flrt-2蛋白表達(dá)增加,N-cadherin蛋白表達(dá)則下降。[結(jié)論]1.鼻中隔軟骨細(xì)胞糖胺聚糖表達(dá)及軟骨結(jié)節(jié)形成能力均高于髁突軟骨細(xì)胞,加載靜磁場(chǎng)后兩種軟骨細(xì)胞糖胺聚糖表達(dá)及軟骨結(jié)節(jié)形成能力均較空白組高。2.鼻中隔軟骨細(xì)胞增殖和遷移能力均強(qiáng)于髁突軟骨細(xì)胞,加載靜磁場(chǎng)后兩種軟骨細(xì)胞增殖和遷移能力均增強(qiáng)。3.軟骨細(xì)胞凝集期,Flrt-2蛋白表達(dá)下調(diào),N-cadherin上調(diào);凝集期過(guò)后Flrt-2蛋白表達(dá)上調(diào),N-cadherin下調(diào)。加載靜磁場(chǎng)后,兩種軟骨細(xì)胞Flrt-2蛋白表達(dá)增加,N-cadherin蛋白表達(dá)則下降。
[Abstract]:[objective] to establish the model of primary mandibular condylar chondroc-chondroc-chondroc (Condylar) and nasal septal chondrocyte (NSCC) cultured in neonatal SD rats under the action of magnetostatic magnetic field, and to observe the effect and difference of magnetic field on cartilage formation of condylar chondrocytes and nasal septal chondrocytes. In order to further study the effect of Flrt-2 on the formation of condylar cartilage and nasal septal cartilage and provide guidance for further study of the effect of magnetostatic magnetic field on the growth and development of maxillofacial region, the theoretical basis is provided for the further study of the effect of magnetostatic magnetic field on the growth and development of maxillofacial region. [methods] the condylar cartilage and nasal septal cartilage of SD rats were taken out for 3 days. Primary cells were obtained by mechanical separation and enzyme-linked digestion. Primary culture and chondrocyte identification were carried out under the same culture conditions. The effect of 280mT static magnetic field on the expression of glycosaminoglycan Gage in two kinds of chondrocytes was detected by acinyl blue staining, and the growth curve of chondrocytes was described by CCK-8(Cell Counting Kit-8 CCK-8 colorimetry. The effect of static magnetic field on the proliferation of two chondrocytes was examined, and the migration ability of the two cells after 24 hours of scratch was observed by scratch test. The expression of Flrt-2 and N-cadherin protein in chondrocytes was detected by Western blotblotWB after static magnetic field loading. [results] the primary condylar chondrocytes and nasal septal chondrocytes were successfully cultured in the same digestion mode and culture conditions. The two kinds of chondrocytes were divided into blank group and magnetic field group. After most of the cells were adhered to the wall, the magnetic field of the cells loaded with 280mT in the magnetic field group continued to grow. The four groups of chondrocytes were stained with acinyl blue on the 1st and 3rd day. The results showed that the expression of glycosaminoglycan and the ability of forming chondrocytes in nasal septal chondrocytes were higher than those in condylar chondrocytes. The expression of glycosaminoglycan and the ability of forming chondrocytes in the two groups were higher than those in the blank group, and the growth curves of the two chondrocytes were similar by CCK-8 colorimetry. The proliferation ability of nasal septal chondrocytes was stronger than that of condylar chondrocytes. The results of 24 h scratch test showed that the migration ability of nasal septal chondrocytes was stronger than that of condylar chondrocytes. The migration ability of the cells in the magnetic field group was stronger than that in the blank group. The expression of Flrt-2N-cadherin protein was detected on the 8th day of culture in two kinds of cells. It was found that the expression of Flrt-2 protein was down-regulated in chondrocyte agglutination phase, and the expression of Flrt-2 protein was up-regulated after the agglutination phase, and the expression of N-cadherin was down-regulated. The expression of Flrt-2 protein in the two chondrocytes increased and the expression of N-cadherin decreased after static magnetic field loading. [conclusion] 1. The expression of glycosaminoglycan and the ability of forming cartilage nodules in nasal septal chondrocytes were higher than those in condylar chondrocytes. The proliferation and migration ability of nasal septal chondrocytes was stronger than that of condylar chondrocytes. The expression of Flrt-2 protein in chondrocytes was down-regulated and the expression of Flrt-2 protein was up-regulated after the agglutination stage. The expression of Flrt-2 protein in the two chondrocytes increased and the expression of N-cadherin decreased after static magnetic field loading.
【學(xué)位授予單位】：昆明醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R782

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 馮其帥;高麗娜;崔元璐;;ATDC5:一株反映軟骨形成完整過(guò)程的細(xì)胞系[J];天津中醫(yī)藥大學(xué)學(xué)報(bào);2015年06期

2 王勝國(guó);周力;陳揚(yáng)熙;;不同強(qiáng)度靜磁場(chǎng)對(duì)成骨細(xì)胞細(xì)胞骨架改建的影響[J];中國(guó)組織工程研究與臨床康復(fù);2011年15期

3 王勝國(guó);周力;陳揚(yáng)熙;王艷民;任曉華;;不同強(qiáng)度靜磁場(chǎng)對(duì)成骨細(xì)胞細(xì)胞內(nèi)鈣離子濃度的影響[J];第三軍醫(yī)大學(xué)學(xué)報(bào);2010年23期

4 周建;陳克明;葛寶豐;李志鋒;程國(guó)政;韋哲;明磊國(guó);翟遠(yuǎn)坤;朱瑞清;韓桂秋;;不同強(qiáng)度正弦交變磁場(chǎng)對(duì)體外培養(yǎng)成骨細(xì)胞增殖與分化影響[J];中國(guó)骨質(zhì)疏松雜志;2010年07期

5 張璐;王沂;胡俊;翁新楚;;超強(qiáng)靜磁場(chǎng)對(duì)小鼠抗氧化和免疫功能的影響[J];上海大學(xué)學(xué)報(bào)(自然科學(xué)版);2009年02期

6 齊浩;艾霞;孫潤(rùn)廣;劉穎;郭偉;;AFM觀察穩(wěn)恒磁場(chǎng)對(duì)細(xì)胞表面精細(xì)結(jié)構(gòu)的影響[J];電子顯微學(xué)報(bào);2009年02期

7 宋國(guó)麗;蘇海靜;張小云;;不同強(qiáng)度的靜磁場(chǎng)對(duì)K562細(xì)胞的作用[J];中國(guó)康復(fù)醫(yī)學(xué)雜志;2009年03期

8 胡麗芳;騫愛(ài)榮;楊鵬飛;張維;謝麗;商澎;;中等強(qiáng)度靜磁場(chǎng)對(duì)白血病細(xì)胞增殖和細(xì)胞周期的影響[J];第四軍醫(yī)大學(xué)學(xué)報(bào);2009年05期

9 朱琳琳;仇麗鴻;;靜磁場(chǎng)對(duì)鼠成骨細(xì)胞增殖和胞內(nèi)鈣離子濃度的影響[J];實(shí)用口腔醫(yī)學(xué)雜志;2008年03期

10 張海軍;吳華;劉偉軍;郝�；�;趙文春;;工頻磁場(chǎng)對(duì)骨髓間充質(zhì)干細(xì)胞基因表達(dá)譜的影響[J];中華物理醫(yī)學(xué)與康復(fù)雜志;2007年04期

相關(guān)博士學(xué)位論文 前1條

1 王麗艷;靜磁場(chǎng)在口腔正畸臨床中的應(yīng)用以及對(duì)面領(lǐng)骨骼肌細(xì)胞生物安全性的研究[D];第四軍醫(yī)大學(xué);2010年

，

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