本文選題：成骨樣細胞MG63 + 壓應(yīng)力��； 參考：《吉林大學(xué)》2014年碩士論文

【摘要】：目的與背景: 牙槽骨是骨組織中可塑性最強、代謝最活躍者。生理情況下，有牙牙合牙槽骨保持破骨與成骨的動態(tài)平衡，使骨組織處于相對穩(wěn)定的狀態(tài)。長期缺牙的患者，牙槽骨缺乏牙齒的功能性刺激，而發(fā)生進行性、累積性、不可逆轉(zhuǎn)的骨吸收。牙槽骨的不斷吸收是引起臨床很多修復(fù)體失敗的主要原因。應(yīng)力因素是影響剩余牙槽嵴吸收的主要因素，也是我們?nèi)藶榈哪軌虿扇「深A(yù)措施加以改變的。王景云等學(xué)者發(fā)現(xiàn)，應(yīng)力與牙槽骨的吸收密切相關(guān)；咀嚼壓力過大會加速剩余牙槽嵴的吸收；適宜的咀嚼壓力能夠延緩剩余牙槽嵴的吸收。 Sato等學(xué)者從人胚胎cDNA文庫中發(fā)現(xiàn)了一條新基因，即：膜型基質(zhì)金屬蛋白酶-1（MT1-MMP），其表達于細胞膜表面，能夠編碼含582個氨基酸的蛋白質(zhì)。近年來研究發(fā)現(xiàn)，作為MMPs家族中被發(fā)現(xiàn)的第一個膜型MMPs，MT1-MMP在正常骨骼發(fā)育及骨改建過程中具有重要的作用。MT1-MMP表達于成骨細胞和破骨細胞，不僅其自身與正常骨骼組織發(fā)育及骨改建有著密切的聯(lián)系，尚可通過激活其他MMPs及細胞因子而影響骨改建的過程；MT1-MMP能夠調(diào)控細胞的機械敏感性；在成骨細胞分化為骨細胞的過程中，MT1-MMP作為一種骨細胞生成初期的必須物質(zhì)具有決定性作用。研究MT1-MMP在骨組織中的作用形式及作用途徑，對了解及掌握骨組織相關(guān)的疾患及研究防治方法具有重要的指導(dǎo)意義。 本實驗運用四點彎曲力學(xué)加載儀對體外培養(yǎng)的MG-63細胞施加不同強度及不同作用時間的壓應(yīng)力，利用Western Blot及逆轉(zhuǎn)錄PCR檢測MT1-MMP的表達水平，為研究MT1-MMP在成骨細胞中的作用機制提供了基礎(chǔ)實驗依據(jù)，進一步證實應(yīng)力作用下的細胞將發(fā)生的細胞生物學(xué)的改變以及探索咀嚼壓力與剩余牙槽嵴吸收的關(guān)系奠定理論基礎(chǔ)。 實驗方法： 采用四點彎曲力學(xué)加載儀對培養(yǎng)在細胞加力板上的MG63細胞施加壓應(yīng)力，頻率為0.5Hz，設(shè)置梯度變化及單一水平的壓應(yīng)力加載方式，研究壓應(yīng)力在不同力值（0、1000μstrain、2000μstrain、4000μstrain、6000μstrain）及不同時間（0、1h、3h、6h、9h、12h）對MT1-MMP蛋白及基因表達的影響，每組實驗重復(fù)3次，空白對照組不加力；除應(yīng)力因素外，其他影響因素基本控制一致。 結(jié)果： 1、當(dāng)應(yīng)變值恒定為2000μstrain時，力學(xué)刺激呈時間依賴性的上調(diào)成骨樣細胞MG63細胞中MT1-MMP蛋白及基因的表達； 2、當(dāng)應(yīng)變值為2000μstrain時，力學(xué)刺激對成骨樣細胞MG63中MT1-MMP蛋白及基因表達作用最強，當(dāng)應(yīng)變值為6000μstrain時，MT1-MMP表達最低。 結(jié)論： 1、在生理范圍內(nèi)，，當(dāng)對MG63細胞施加恒定的壓應(yīng)力時，MT1-MMP的表達呈時間依賴性的上調(diào)； 2、當(dāng)應(yīng)力加載時間相同時，MT1-MMP的表達隨著力值的增大而增加，但應(yīng)力超出生理范圍時，其表達則降低。 3、適宜的應(yīng)力在調(diào)節(jié)骨改建過程中有著重要的作用。
[Abstract]:Purpose and background: Alveolar bone is the most plastic and metabolized bone tissue. Under physiological conditions, the alveolar bone of edentulous teeth keeps the dynamic balance between osteoclast and osteogenesis, which makes the bone tissue relatively stable. In patients with chronic dental deficiency, alveolar bone lacks functional stimulation and develops progressive, cumulative, irreversible bone resorption. The incessant absorption of alveolar bone is the main cause of the failure of many clinical prostheses. Stress factor is the main factor affecting the absorption of residual alveolar ridge. Wang Jingyun and other scholars found that stress is closely related to alveolar bone absorption; masticatory pressure is excessive to accelerate the absorption of residual alveolar ridge; appropriate masticatory pressure can delay the absorption of residual alveolar ridge. Sato and other researchers have found a new gene from the human embryo cDNA library, namely, the membrane matrix metalloproteinase-1 (MMP-1) MT1-MMPN, which is expressed on the surface of the cell membrane and encodes a protein containing 582 amino acids. In recent years, it has been found that MT1-MMP, the first membrane type of MMPsMMP found in MMPs family, plays an important role in normal bone development and bone remodeling. MT1-MMP is expressed in osteoblasts and osteoclasts. Not only is it closely related to normal bone tissue development and bone remodeling, but it can also affect the process of bone remodeling by activating other MMPs and cytokines. MT1-MMP can regulate the mechanical sensitivity of cells. MT1-MMP plays a decisive role in the process of osteoblast differentiation into osteocytes. It is of great significance to understand and master the diseases related to bone tissue and to study the methods of prevention and treatment by studying the forms and ways of action of MT1-MMP in bone tissue. In this study, four point bending mechanical loading apparatus was used to apply compressive stress of different intensity and time to MG-63 cells in vitro. The expression of MT1-MMP was detected by Western Blot and reverse transcription PCR. It provides a basic experimental basis for studying the mechanism of MT1-MMP acting on osteoblasts, further confirms the changes of cell biology under stress and explores the relationship between masticatory pressure and residual alveolar ridge absorption. Experimental methods: A four-point bending mechanical loading instrument was used to apply compressive stress to the MG63 cells cultured on the cell afterloading plate at a frequency of 0.5 Hz, and a gradient change and a single level of compressive stress loading mode were set. The effects of compressive stress on the expression of MT1-MMP protein and gene were studied at different stress values (0 1 000 渭 strain (2000 渭 strain) and 4 000 渭 strain) and at different times. The results showed that the MT1-MMP protein and gene expression were repeated 3 times in each group, but not in the blank control group. Except stress factors, the other influencing factors were basically controlled. Results: 1. When the strain value was 2000 渭 strain, mechanical stimulation up-regulated the expression of MT1-MMP protein and gene in MG63 cells in a time-dependent manner. 2. When the strain value was 2000 渭 strain, the expression of MT1-MMP protein and gene in osteoblast like cell MG63 was the strongest by mechanical stimulation, and the expression of MT1-MMP was the lowest when the strain value was 6000 渭 strain. Conclusion: 1. In physiological range, the expression of MT1-MMP was up-regulated in a time-dependent manner when MG63 cells were subjected to constant compressive stress. 2. When the stress loading time is the same, the expression of MT1-MMP increases with the increase of stress, but when the stress exceeds the physiological range, the expression of MT1-MMP decreases. 3. Suitable stress plays an important role in regulating bone remodeling.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R783

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