本文關(guān)鍵詞： 骨折愈合 成骨細(xì)胞 持續(xù)與間歇低載荷振動(dòng)　出處：《吉林大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：研究背景和目的:隨著全球經(jīng)濟(jì)的飛速發(fā)展,交通事故傷與高處墜落傷等高能量損傷病人逐年增多,此類損傷已成為和平時(shí)期創(chuàng)傷發(fā)生的最常見(jiàn)原因。在以上損傷中,四肢損傷,特別是四肢骨折的發(fā)生率極高,約有5%-10%患者會(huì)發(fā)生骨折延遲愈合或骨不連,嚴(yán)重影響了骨折的康復(fù)進(jìn)程。因此,怎樣加快骨折愈合,有效防治骨折延遲愈合或不愈合已經(jīng)成為熱門課題。近十年來(lái)的研究表明,低載荷(頻率32-37Hz,加速度0.2-0.3g)振動(dòng)(low-magnitude high-frequency vibration LMHFV)能夠促進(jìn)骨的生長(zhǎng)、對(duì)抗骨質(zhì)疏松、促進(jìn)骨折愈合。然而絕大多數(shù)文獻(xiàn)報(bào)導(dǎo)的振動(dòng)對(duì)骨及骨細(xì)胞的影響均是采用持續(xù)的振動(dòng)加載方式鮮有文獻(xiàn)報(bào)導(dǎo)研究持續(xù)與間歇振動(dòng)對(duì)骨及骨細(xì)胞是否會(huì)有差別。因此本實(shí)驗(yàn)擬通過(guò)對(duì)體外培養(yǎng)的MC3T3-E1細(xì)胞施加持續(xù)低載荷振動(dòng)(sostenuto s LMHFV)與間歇低載荷振動(dòng)(intermittent LMHFV i LMHFV)兩種不同的低載荷振動(dòng)加載模式,通過(guò)細(xì)胞層面的研究來(lái)探索持續(xù)與間歇低載荷振動(dòng)對(duì)骨折愈合是否會(huì)產(chǎn)生不同的影響及其作用機(jī)理。研究方法:使用振動(dòng)加載設(shè)備,對(duì)MC3T3-E1細(xì)胞施加振幅35Hz,強(qiáng)度0.25g的低載荷機(jī)械振動(dòng)。分成空白對(duì)照組,持續(xù)振動(dòng)0.5h組,振動(dòng)15min間歇0.5h、1h、3h、6h后再振動(dòng)15min組。選取振動(dòng)加載的第1、3、5、7天進(jìn)行MTT檢測(cè),來(lái)測(cè)定MC3T3-E1細(xì)胞的增殖活性。連續(xù)三天低載荷振動(dòng)加載結(jié)束后運(yùn)用ALP試劑盒檢測(cè)胞內(nèi)ALP活性,運(yùn)用Western-blot、RT-q PCR技術(shù)來(lái)檢測(cè)低載荷振動(dòng)刺激對(duì)成骨細(xì)胞ALP、OCN、COLⅠ、OPG、RANKL等因子的表達(dá)水平,運(yùn)用ELISA試劑盒檢測(cè)MC3T3-E1細(xì)胞成骨細(xì)胞分泌RANKL及M-CSF兩種破骨細(xì)胞刺激因子的情況。實(shí)驗(yàn)結(jié)果:不論在哪一天檢測(cè),持續(xù)與間歇低載荷振動(dòng)組的細(xì)胞增殖活性與對(duì)照組無(wú)明顯差別,持續(xù)低載荷振動(dòng)組的細(xì)胞增殖活性與間歇低載荷振動(dòng)組的細(xì)胞增殖活性也沒(méi)有明顯差別。所有振動(dòng)組的MC3T3-E1細(xì)胞的ALP、OCN、COLⅠ的表達(dá)量及OPG/RANKL比值均明顯優(yōu)于對(duì)照組,并且間歇0.5h振動(dòng)組較持續(xù)振動(dòng)組效果更好。低載荷振動(dòng)抑制了MC3T3-E1細(xì)胞分泌RANKL及M-CSF因子,并且間歇0.5h振動(dòng)組、間歇1h振動(dòng)組兩組的RANKL及M-CSF的分泌量較持續(xù)振動(dòng)組受到的更加明顯的抑制。結(jié)論:低載荷振動(dòng)(35Hz,0.25g)可以促進(jìn)成骨細(xì)胞的分化,并且可抑制成骨細(xì)胞分泌破骨細(xì)胞刺激因子,進(jìn)而抑制破骨細(xì)胞活性;間隔0.5h的低載荷振動(dòng)促進(jìn)成骨細(xì)胞分化的效果優(yōu)于持續(xù)振動(dòng)組;低載荷振動(dòng)對(duì)成骨細(xì)胞的增殖無(wú)顯著影響。
[Abstract]:Background and objective: with the rapid development of the global economy, the number of patients with high-energy injuries, such as traffic accidents and falling injuries, has increased year by year, and such injuries have become the most common cause of peacetime trauma. The incidence of limb injuries, especially limb fractures, is extremely high. About 5- 10 percent of the patients have delayed union or nonunion, which seriously affects the recovery process of fractures. Therefore, how to accelerate fracture healing, The effective prevention and treatment of delayed healing or nonunion of fracture has become a hot topic. Recent studies have shown that low-load vibration (frequency 32-37Hz, acceleration 0.2-0.3 g) can promote bone growth and combat osteoporosis. However, the effect of vibration on bone and bone cells is mostly reported in the literature. It is rarely reported that there is a difference between continuous vibration and intermittent vibration on bone and bone cells. In this study, two different modes of low load vibration loading were proposed for MC3T3-E1 cells: continuous low load vibration (Sostenuto s LMHFV) and intermittent low load vibration LMHFV I (LMHFV). Through cell-level studies to explore whether sustained and intermittent low-load vibration has different effects on fracture healing and its mechanism. The MC3T3-E1 cells were subjected to low load mechanical vibration with amplitude of 35 Hz and intensity of 0.25 g. The cells were divided into blank control group, continuous vibration group for 0.5 h, vibration interval for 15 minutes for 0.5 h and 1 h for 3 h and then 15 min group for 15 min. The MTT was detected on the 5th day of vibration loading. After three days of low load vibration loading, ALP kit was used to detect intracellular ALP activity, Western-blottRT-q PCR technique was used to detect the expression level of factors such as ALP OCNCOL 鈪，

本文編號(hào)：1532085