【摘要】：目的 通過(guò)兔骨髓間質(zhì)干細(xì)胞在PRF膜上增殖和分化的實(shí)驗(yàn)，研究PRF膜在誘導(dǎo)骨髓間質(zhì)干細(xì)胞向成骨細(xì)胞轉(zhuǎn)化的作用。為PRF膜應(yīng)用于口腔種植學(xué)領(lǐng)域提供分子生物學(xué)的依據(jù)。 方法 取7天生新西蘭大白兔股骨和脛骨，分離骨髓間質(zhì)干細(xì)胞進(jìn)行體外培養(yǎng)，在各自的培養(yǎng)條件下分為三組：空白對(duì)照組（MSCs）、實(shí)驗(yàn)組（PRF膜+MSCS)和陽(yáng)性對(duì)照組（BMP-2+MSCs）。其中，實(shí)驗(yàn)組根據(jù)PRF膜劑量的不同，分為五個(gè)實(shí)驗(yàn)亞組，分別是PRF膜1ml組，2ml組,3ml組,4ml組和5ml組。三組細(xì)胞分別在24h，48h，72h采用MTT法測(cè)定細(xì)胞增殖率、在72h用PNPP法測(cè)定堿性磷酸酶(ALP)的表達(dá)、免疫組化檢測(cè)Ⅰ型膠原蛋白，骨鈣素的表達(dá)。通過(guò)以上方法檢測(cè)細(xì)胞的成骨特性。 結(jié)果 細(xì)胞形態(tài)學(xué)觀察，MSCS分化后，細(xì)胞形態(tài)從長(zhǎng)梭形變成三角形，多角形，立方形；MTT測(cè)定細(xì)胞增殖顯示，實(shí)驗(yàn)組五個(gè)亞組呈劑量反應(yīng)關(guān)系（即隨著PRF膜劑量的增加，細(xì)胞增殖數(shù)量增加）與空白對(duì)照組比較，各實(shí)驗(yàn)亞組具有統(tǒng)計(jì)學(xué)意義（F=89.548，P=0.000）；與陽(yáng)性對(duì)照組比較，PRF膜劑量為1ml，2ml和3ml時(shí)，細(xì)胞增殖數(shù)量隨著PRF膜劑量的增大而增加，但均小于陽(yáng)性對(duì)照組；PRF膜劑量為4ml和5ml時(shí)，，與陽(yáng)性對(duì)照組差別不大，無(wú)統(tǒng)計(jì)學(xué)意義（F=3.364，P=0.142）；ALP檢測(cè)結(jié)果顯示，MSCS分化后，實(shí)驗(yàn)組五個(gè)亞組呈劑量反應(yīng)關(guān)系（即隨著PRF膜劑量的增加，ALP活性增強(qiáng)）與空白對(duì)照組比較，各實(shí)驗(yàn)亞組具有統(tǒng)計(jì)學(xué)意義。與陽(yáng)性對(duì)照組比較，PRF膜劑量為1ml，2ml和3ml時(shí)，ALP活性隨著PRF膜劑量的增大而增強(qiáng)，但均小于陽(yáng)性對(duì)照組。PRF膜劑量為4ml和5ml時(shí)，與陽(yáng)性對(duì)照組差別不大，無(wú)統(tǒng)計(jì)學(xué)意義。免疫組化標(biāo)記空白對(duì)照組，PRF膜4ml組，陽(yáng)性對(duì)照組的Ⅰ型膠原蛋白和骨鈣素，結(jié)果均顯示陽(yáng)性。即棕褐色顆粒沉著，陽(yáng)性顆粒出現(xiàn)在細(xì)胞漿，細(xì)胞膜和細(xì)胞質(zhì)基質(zhì)。提示，細(xì)胞處于Ⅰ型膠原；骨鈣素合成分泌的不同時(shí)期。推測(cè)，PRF膜5ml組細(xì)胞也會(huì)具有相同的陽(yáng)性反應(yīng)。 結(jié)論 PRF膜可以誘導(dǎo)骨髓間質(zhì)干細(xì)胞分化為成骨細(xì)胞，分化的細(xì)胞具有成骨細(xì)胞的特性，可以作為自體材料應(yīng)用于口腔種植學(xué)領(lǐng)域里骨缺損的修復(fù)。
[Abstract]:Objective to investigate the effect of PRF membrane on the proliferation and differentiation of rabbit bone marrow mesenchymal stem cells (BMSCs) into osteoblasts. It provides molecular biology basis for the application of PRF membrane in the field of stomatology. Methods Bone marrow mesenchymal stem cells (BMSCs) were isolated from femur and tibia of 7 born New Zealand white rabbits and cultured in vitro. They were divided into three groups: blank control group (MSCs),). Experimental group (PRF membrane MSCS) and positive control group (BMP-2 MSCs).) According to the dosage of PRF membrane, the experimental group was divided into five subgroups: PRF membrane 1ml group, 2ml group, 3ml group, 4ml group and 5ml group. The proliferation rate of the three groups was measured by MTT assay at 24 h and 48 h at 72 h, the expression of alkaline phosphatase (ALP) by PNPP method at 72 h, and the expression of type I collagen and osteocalcin by immunohistochemistry. The osteogenic characteristics of the cells were detected by the above methods. Results after MSCS differentiation, cell morphology changed from long spindle to triangle, polygonal and square. MTT assay showed that there was a dose-response relationship among the five subgroups in the experimental group (that is, with the increase of the PRF membrane dose, the number of cell proliferation increased), which was significantly different from that of the control group (F _ (89.548) P _ (0.000). Compared with the positive control group, the number of cell proliferation increased with the increase of PRF membrane dose when the PRF membrane dose was 1ml and 3ml, but both were smaller than those of the positive control group. When the dosage of PRF membrane was 4ml and 5ml, there was no significant difference between the positive control group and the 4ml membrane dose (FN 3.364 P 0.142). The results of ALP showed that after MSCS differentiation, the five subgroups of the experimental group showed dose-response relationship (that is, with the increase of PRF membrane dose, the activity of ALP increased), compared with the control group, the experimental subgroups had statistical significance. Compared with the positive control group, the activity of ALP increased with the increase of PRF membrane dose when the PRF membrane dose was 1ml / L and 3ml, but they were smaller than those of the positive control group. When the PRF membrane dosage was 4ml and 5ml, the difference was not significant between the positive control group and the positive control group, and there was no significant difference between the two groups. Immunohistochemical staining showed positive expression of type I collagen and osteocalcin in the blank control group, PRF membrane 4ml group and positive control group. That is, brown granules, positive granules appear in the cytoplasm, cell membrane and cytoplasmic matrix. These results suggest that the cells are at different stages of osteocalcin synthesis and secretion. It is speculated that the cells in PRF membrane 5ml group also have the same positive reaction. Conclusion PRF membrane can induce bone marrow mesenchymal stem cells to differentiate into osteoblasts. The differentiated cells have the characteristics of osteoblasts and can be used as autogenous materials to repair bone defects in the field of dental implants.
【學(xué)位授予單位】：遼寧醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R329

【相似文獻(xiàn)】

相關(guān)期刊論文 前10條

1 張麗蓉,夏文杰,項(xiàng)鵬,陳振光,張秀明,李艷,李樹(shù)濃;體外定向誘導(dǎo)人骨髓間質(zhì)干細(xì)胞分化為成骨細(xì)胞的研究[J];中國(guó)病理生理雜志;2002年07期

2 毛磊;;骨氃干細(xì)胞可修復(fù)心臟損傷[J];科學(xué)咨詢(決策管理);2003年11期

3 孫曉楠,劉國(guó)樹(shù),石蕊,龐偉,李玉明;骨髓間質(zhì)干細(xì)胞移植對(duì)鈣化主動(dòng)脈功能及骨橋蛋白表達(dá)水平的影響[J];中國(guó)藥物應(yīng)用與監(jiān)測(cè);2005年04期

4 田瑩;鄧宇斌;那曉東;;骨髓間質(zhì)干細(xì)胞在造血干細(xì)胞移植中的作用研究進(jìn)展[J];國(guó)際輸血及血液學(xué)雜志;2005年06期

5 朱志勇;周慶國(guó);魏毅東;魏經(jīng)漢;;骨髓間質(zhì)干細(xì)胞靜脈移植治療大鼠急性心肌梗死的實(shí)驗(yàn)研究[J];臨床心血管病雜志;2006年03期

6 張培全;曾和平;;5-[2-(8-羥基喹啉-2-基)乙烯基]-2-甲基-8-羥基喹啉的合成、抗氧化活性及促進(jìn)鼠骨髓間質(zhì)干細(xì)胞增殖的研究[J];有機(jī)化學(xué);2008年06期

7 張培全;曾和平;;乙烯基橋聯(lián)的8-羥基喹啉衍生物的合成、抗氧化活性及促進(jìn)鼠骨髓間質(zhì)干細(xì)胞增殖的研究[J];高等學(xué)校化學(xué)學(xué)報(bào);2010年01期

8 劉凱;吳建軍;;鎖陽(yáng)含藥血清對(duì)骨髓間質(zhì)干細(xì)胞向神經(jīng)細(xì)胞分化的影響[J];中醫(yī)研究;2010年05期

9 黃鈾新;呂富榮;呂發(fā)金;馬秀華;肖云華;肖智博;;CT腦灌注劑量下X射線對(duì)體外培養(yǎng)的骨髓間質(zhì)干細(xì)胞的影響[J];生命科學(xué)研究;2010年06期

10 李正;吳文;王tR;王小娜;;雌激素在骨髓間質(zhì)干細(xì)胞轉(zhuǎn)化為成骨細(xì)胞中的作用[J];廣東醫(yī)學(xué);2011年11期

相關(guān)會(huì)議論文 前10條

1 張建保;M Bratbach;BK Fleischmann;;骨髓間質(zhì)干細(xì)胞電壓依賴性離子通道[A];第七屆全國(guó)生物力學(xué)學(xué)術(shù)會(huì)議論文集[C];2003年

2 周小瑩;暨明;王先酉;田晶;王峰;趙菲;譚孟群;;rAAV2介導(dǎo)TPO基因修飾的骨髓間質(zhì)干細(xì)胞對(duì)巨核系細(xì)胞生成的促進(jìn)作用[A];湖南省生理科學(xué)會(huì)2006年度學(xué)術(shù)年會(huì)論文摘要匯編[C];2007年

3 陳少?gòu)?qiáng);吳碧蓮;賈小力;;移植骨髓間質(zhì)干細(xì)胞在損傷脊髓內(nèi)向神經(jīng)元的定向分化[A];中國(guó)解剖學(xué)會(huì)2011年年會(huì)論文文摘匯編[C];2011年

4 景黎君;賈永林;魯晶晶;韓瑞;王舒陽(yáng);李盡義;彭濤;賈延R

本文編號(hào)：2333046