本文選題：堿熱處理 + 鈦片表面�。� 參考：《昆明醫(yī)科大學》2014年碩士論文

【摘要】：[目的]研究堿熱處理的鈦片表面對成骨細胞整合素α2β1,整合素α5β1的影響。 [材料和方法]本實驗用SD胎鼠顱頂骨組織塊培養(yǎng)法進行成骨細胞原代培養(yǎng)。用倒置顯微鏡觀察原代成骨細胞的生長情況,并利用HE染色、堿性磷酸酶染色及鈣化結節(jié)茜素紅染色進行成骨細胞鑒定。將直徑為15mm,厚lmm的醫(yī)用純鈦鈦片分組進行表面處理：①以240目、360目、600目砂紙逐級打磨得到機械打磨組(grooved,G組)；②采用356-411μmTiO2噴砂處理,得到噴砂組(sandblasted,SB組)；③以63.6%的H2S04和10.6%的HCL混合液在60。C時酸蝕SB組的鈦片10分鐘得到酸蝕組(sandblasted and acid-etched,SLA組)；④由G組、SB組、SLA組,經5moL/L氫氧化鈉60。C水浴14小時,600。C處理1小時,得到光滑堿熱組(grooved alkali-heated,AH1組)、噴砂堿熱組(sandblasted alkali-heated,AH2組)、噴砂酸蝕堿熱組(acid-etched and alkali-heated,AH3組)。用掃描電鏡、輪廓測量儀檢測6組鈦片表面理化性質。將成骨細胞接種于六組鈦片上,在1、3、5天收集所培養(yǎng)的成骨細胞。利用RT-PCR定量免疫熒光法檢測成骨細胞整合素亞基α2、整合素亞基a5、整合素亞基β1mRNA的表達水平。數(shù)據(jù)采用單因素方差分析及配對t檢驗(a=0.05)。 [結果]本實驗采用的體外組織塊法操作簡便,骨量消耗小,細胞產量大、純度高,能滿足實驗需求。培養(yǎng)的成骨細胞經鑒定符合成骨細胞的生物學特性,并具有良好的活性。體外培養(yǎng)的大鼠成骨細胞膜上均能持續(xù)表達整合素亞基α2、整合素亞基α5、整合素亞基β1。整合素亞基p1表達水平最高,其次為整合素亞基α2,整合素亞基α5相對最低。本實驗中,SB組鈦片表面處理后成骨細胞整合素表達水平最高,AH1組較G組、AH3組較SLA組的表達水平要高,AH1組、AH1組和AH3組表達水平接近。在本實驗觀察時段中,隨著時間的延長,整合素亞基α2、整合素亞基α5、整合素亞基β1mRNA的表達水平有一定程度的增長。 [結論]1.實驗采用組織塊法培養(yǎng)成骨細胞,操作簡便,骨量消耗小,細胞產量大、純度高,能滿足實驗需求。獲得的成骨細胞經鑒定符合成骨細胞的生物學特性,并具有良好的活性。2.實驗引物特異性佳,擴增效率高,成功檢測到3種目標整合素亞基,擴增出相應目的片段,合乎實驗要求。3.SB組鈦片成骨細胞整合素亞基a2、整合素亞基a5、整合素亞基p1表達量最大,SB組有利于成骨細胞粘附。4.堿熱處理表面對成骨細胞整合素亞基a2、整合素亞基α5、整合素亞基β1表達有一定促進作用,有利于成骨細胞粘附。
[Abstract]:[objective] to study the effect of alkaline heat treated titanium sheet on integrin 偽 2 尾 1 and integrin 偽 5 尾 1 in osteoblasts. Materials and methods the primary culture of osteoblasts was carried out by the method of SD fetal rat skullcap tissue mass culture. The growth of primary osteoblasts was observed by inverted microscope. Osteoblasts were identified by HE staining, alkaline phosphatase staining and alizarin red staining of calcified nodules. The titanium sheet with a diameter of 15 mm and thickness of lmm was divided into two groups. The surface treatment was carried out with 240 mesh / 360 mesh / 600 mesh sandpaper, and the mechanical grinding group was treated with 356-411 渭 mTiO2 sandblasting, and the mechanical grinding group was treated with 356-411 渭 mTiO2. The sand-blast group and blasted SB group were treated with 63.6% H2S04 and 10.6% HCL mixture solution for 10 minutes at 60.C, and then treated with 5moL/L sodium hydroxide 60.C for 14 hours and 600.C, respectively, and then treated with 5moL/L sodium hydroxide 60.C for 14 hours or 600.C, then treated with 5moL/L sodium hydroxide for 14 hours or 600.C. The smooth alkali-hested AH1 group, sandblasted alkali-hested AH2 group and acid-etched and alkali-heaved AH3 group were obtained in the smooth alkali fever group, sandblasted alkali-hested AH _ 2 group and sandblasted alkali-heated AH _ 2 group. The surface physicochemical properties of 6 groups of titanium sheets were examined by scanning electron microscope (SEM) and profilometer. Osteoblasts were inoculated on six groups of titanium slices and cultured osteoblasts were collected for 5 days. The expression levels of integrin subunit 偽 2, integrin subunit a5 and integrin subunit 尾 1mRNA in osteoblasts were detected by RT-PCR quantitative immunofluorescence assay. The data were analyzed by single factor ANOVA and paired t test. [results] the method of tissue mass in vitro was simple, the bone mass consumption was small, the cell yield was large, the purity was high, and it could meet the needs of the experiment. The cultured osteoblasts were confirmed to be in accordance with the biological characteristics of osteoblasts and had good activity. In vitro cultured rat osteoblasts, integrin subunit 偽 2, integrin subunit 偽 5 and integrin subunit 尾 1 were continuously expressed. The expression level of integrin subunit p1 was the highest, followed by integrin subunit 偽 2, and integrin subunit 偽 5 was the lowest. In this experiment, the expression level of integrin in osteoblasts of group B was higher than that of group G, AH3 and SLA. The expression level of integrin in osteoblasts of group B was higher than that of group SLA. The expression level of integrin in group AH1 was higher than that in group SLA. The expression level of integrin in group AH1 was similar to that in group AH3. During the observation period, the expression of integrin subunit 偽 2, integrin subunit 偽 5 and integrin subunit 尾 1mRNA increased to some extent. [conclusion] 1. Osteoblasts were cultured by tissue mass method, which was easy to operate, low bone consumption, high cell yield and high purity. The obtained osteoblasts were confirmed to be in accordance with the biological characteristics of osteoblasts and had good activity. 2. 2. Three target integrin subunits were successfully detected and the corresponding target fragments were amplified. In accordance with the experimental requirements, the osteoblasts in group SB had the highest expression of integrin subunit a2, integrin subunit a5 and integrin subunit p1, and the SB group was in favor of osteoblast adhesion of .4. The surface of alkali heat treatment can promote the expression of integrin subunit a2, integrin subunit 偽 5 and integrin subunit 尾 1 in osteoblasts, which is beneficial to osteoblast adhesion.
【學位授予單位】：昆明醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R783.6

【參考文獻】

相關期刊論文 前5條

1 高長有,李安,封麟先;促進組織生長的聚合物骨架工程材料[J];功能高分子學報;1998年03期

2 牛濤;丁仲鵑;董菲;;不同表面處理鈦片對成骨細胞骨架影響的研究[J];華西口腔醫(yī)學雜志;2007年06期

3 鄧煒,王貽寧,蔣滔,陳群,周彬,程祥榮;仿生溶液法誘導鈦表面形成鈣磷涂層的研究[J];生物醫(yī)學工程學雜志;2002年03期

4 楊志明,余希杰,解慧琪,陳旭,屈藝;不同來源成骨細胞生物學特性的比較研究[J];中華創(chuàng)傷雜志;2001年01期

5 王前，，鐘世鎮(zhèn)，龔文匯，歐陽鈞;鼠顱蓋骨成骨細胞體外培養(yǎng)及ALP免疫組化法鑒別純化[J];中華骨科雜志;1995年06期

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