本文關(guān)鍵詞： 鈦 雙輝離子滲氮 成骨細(xì)胞　出處：《山西醫(yī)科大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：目的:觀察近β鈦合金Ti-3Zr-2Sn-3Mo-25Nb(TLM)雙層輝光離子滲氮改性層表面成骨細(xì)胞早期黏附、增殖、堿性磷酸酶活性、成骨相關(guān)基因的表達(dá)情況,研究TLM滲氮改性層對(duì)成骨細(xì)胞早期功能的影響。方法:近β鈦合金TLM圓片試樣經(jīng)拋光、清洗后平均分成兩組,實(shí)驗(yàn)組試樣采用雙層輝光離子滲氮處理,TLM拋光組作為對(duì)照。將小鼠顱頂前骨細(xì)胞亞克隆14接種于兩組試樣表面,掃描電鏡觀察接種4h后細(xì)胞的黏附形貌;MTT法測(cè)3d、5d、7d細(xì)胞增殖情況;檢測(cè)7d、14d細(xì)胞堿性磷酸酶(alkaline phosphatase,ALP)活性;實(shí)時(shí)熒光定量聚合酶鏈反應(yīng)(quantitative real time,qRT-PCR)檢測(cè)細(xì)胞Runt相關(guān)轉(zhuǎn)錄因子2(Runt-related transcription factor-2,RUNX2)、Ⅰ型膠原α1鏈(typeⅠcollagen alpha 1 chain,COLⅠα1)、骨保護(hù)素(osteoprotegerin,OPG)和核因子-κB受體活化因子配體(receptor activator of nuclear factor-kappaB ligand,RANKL)mRNA的表達(dá)情況。結(jié)果:接種4h,TLM滲氮改性組細(xì)胞黏附形態(tài)良好,有細(xì)絲狀偽足伸出,細(xì)胞間連接緊密;培養(yǎng)3d,實(shí)驗(yàn)組細(xì)胞量(0.277±0.007)明顯高于對(duì)照組(0.249±0.004)(P=0.004)而培養(yǎng)5d和7d兩組細(xì)胞量均未見明顯差異;培養(yǎng)14d,實(shí)驗(yàn)組細(xì)胞ALP活性(173.606±1.884)明顯高于對(duì)照組(162.582±2.426)(P=0.003);滲氮組RUNX2、Ⅰ型膠原α1鏈、骨保護(hù)素mRNA相對(duì)表達(dá)量均顯著高于對(duì)照組(P0.05),RANKLmRNA相對(duì)表達(dá)量顯著低于對(duì)照組(P=0.016)。結(jié)論:近β鈦合金TLM表面采用雙層輝光離子滲氮改性后,改性層促進(jìn)成骨細(xì)胞早期黏附,利于成骨細(xì)胞分化,提高細(xì)胞骨保護(hù)素OPGmRNA表達(dá)量,同時(shí)抑制了RANKLmRNA的表達(dá)。推測(cè)在改性層表面可能更快速形成骨結(jié)合。
[Abstract]:Aim: to observe the early adhesion, proliferation, alkaline phosphatase activity and osteoblast-related gene expression of the surface osteoblasts of Ti-3Zr-2Sn-3Mo-25NbTLM layer modified by double glow plasma nitriding. The effects of TLM nitriding modified layer on the early function of osteoblasts were studied. Methods: after polishing and cleaning, the samples of near 尾 -Ti alloy TLM wafer were divided into two groups. The experimental group was treated with double glow ion nitriding and treated with TLM as control group. The subclone 14 of mouse precranial bone cells was inoculated on the surface of the two groups. The cell adhesion morphology was observed by scanning electron microscope (SEM) and MTT method was used to measure the proliferation of the cells for 3 days and 5 days after inoculation. Alkaline phosphatase (ALP) activity was detected on day 14 after 7 days. Real-time quantitative real quantitative real qRT-PCR detection of the expression of Runt related transcription factor 2G Runt-related transcription factor-2, type 鈪，

本文編號(hào)：1541948