本文選題：骨髓基質(zhì)細(xì)胞　切入點(diǎn)：頜骨　出處：《南京醫(yī)科大學(xué)》2008年碩士論文

【摘要】： 因創(chuàng)傷、感染、腫瘤及先天性疾病等造成的頜骨缺損十分常見(jiàn),目前針對(duì)頜骨缺損的主要修復(fù)方法是自體骨移植,其中以髂骨作為骨供體最為常見(jiàn)。胚胎學(xué)研究表明大部分顱頜面骨發(fā)生于顱神經(jīng)嵴來(lái)源外胚間充質(zhì),而軀干四肢骨來(lái)源于中胚層;從成骨方式而言頜骨為膜內(nèi)成骨,髂骨則為軟骨內(nèi)成骨。大量臨床與動(dòng)物實(shí)驗(yàn)表明截?cái)嘈灶M骨缺損采用頜骨供體比其他部位骨供體成功率更高,主要表現(xiàn)為移植供體存活率更高,遠(yuǎn)期吸收率更低。 這些已有的臨床、實(shí)驗(yàn)室、生物學(xué)研究提示我們?cè)诓煌课还枪撬鑳?nèi)的骨祖細(xì)胞可能具有各自生物學(xué)特性;而關(guān)于骨組織修復(fù)的細(xì)胞學(xué)模型主要來(lái)自長(zhǎng)骨,對(duì)頜骨來(lái)源骨髓細(xì)胞的針對(duì)性研究甚少。為此,我們采用大鼠頜骨與髂骨來(lái)源骨髓基質(zhì)干細(xì)胞(Bone Marrow StromalCells,BMSCs)進(jìn)行體外擴(kuò)增培養(yǎng),通過(guò)相應(yīng)的誘導(dǎo)條件,觀(guān)察這兩種不同來(lái)源的BMSCs各自在細(xì)胞增殖,集落形成,分化成骨,礦化形成及分化成脂方面是否存在差異,希望借此為進(jìn)一步探索頜骨在發(fā)生、修復(fù)上是否具有其獨(dú)特生物學(xué)機(jī)制提供一定的依據(jù)和參考。 第一部分大鼠頜骨與髂骨來(lái)源骨髓基質(zhì)干細(xì)胞的體外培養(yǎng) 目的:從大鼠的頜骨與髂骨骨髓中運(yùn)用全骨髓細(xì)胞貼壁培養(yǎng)法分別培養(yǎng)出兩位點(diǎn)BMSCs,為探討頜骨與髂骨來(lái)源BMSCs的生物學(xué)特性是否具有位點(diǎn)性差異提供細(xì)胞模型。 方法:取4～6周齡150克±10克雄性SD大鼠,首先解剖分離出下頜骨及髂骨,無(wú)菌條件下培養(yǎng)基沖出骨髓細(xì)胞制成細(xì)胞懸液,采用全骨髓貼壁法培養(yǎng)細(xì)胞,相差顯微鏡下觀(guān)察細(xì)胞生長(zhǎng)情況及特點(diǎn)。 結(jié)果:原代細(xì)胞接種24h后鏡下可見(jiàn)瓶底有細(xì)胞貼壁,首次換液去除大部分外周血細(xì)胞后可見(jiàn)散在的梭形細(xì)胞聚集;隨反復(fù)換液,瓶?jī)?nèi)未貼壁細(xì)胞逐步被清除,細(xì)胞生長(zhǎng)形成典型的成纖維細(xì)胞樣-集落形成單位(colony-forming unit-fibroblast,CFU-F)。細(xì)胞首次傳代約需2周左右,傳代細(xì)胞于第6代(髂骨)-第9代(頜骨)前細(xì)胞增殖能力比較旺盛,以后細(xì)胞活力逐漸減弱,出現(xiàn)不同的分化傾向,部分細(xì)胞胞漿中出現(xiàn)空泡及顆粒樣物質(zhì),最終死亡。 結(jié)論:全骨髓細(xì)胞貼壁培養(yǎng)法能夠培養(yǎng)出滿(mǎn)足實(shí)驗(yàn)要求的BMSCs。 第二部分大鼠頜骨與髂骨來(lái)源骨髓基質(zhì)干細(xì)胞體外生物學(xué)特性的比較研究 目的:探討體外環(huán)境下大鼠頜骨與髂骨來(lái)源BMSCs體外生物學(xué)性狀是否存在差異,為進(jìn)一步深入體內(nèi)研究提供依據(jù)。 方法:取第二代正常生長(zhǎng)的頜骨與髂骨BMSCs,分別從細(xì)胞增殖活性,誘導(dǎo)成骨,礦化及成脂能力等多方面比較兩者的體外生物學(xué)特性的差異。 結(jié)果:頜骨來(lái)源的BMSCs與髂骨相比,在低濃度接種條件下具有更強(qiáng)的增殖能力,誘導(dǎo)成骨后更高的堿性磷酸酶陽(yáng)性表現(xiàn),礦化誘導(dǎo)后形成更多的鈣結(jié)節(jié);而髂骨來(lái)源BMSCs在成脂誘導(dǎo)條件下分化形成更多脂質(zhì)。 結(jié)論:頜骨與髂骨來(lái)源BMSCs在體外具有不同的生物學(xué)特點(diǎn),提示頜骨來(lái)源的BMSCs可能具有其獨(dú)特生物學(xué)特性,為進(jìn)一步研究頜骨病變與再生修復(fù)機(jī)制提供實(shí)驗(yàn)依據(jù)和參考。
[Abstract]:The main repair method for jaw defects is autogenous bone grafting , which is the most common bone donor in the bone graft . The study of embryology shows that most of the cranio - maxillofacial bones are derived from the ectodermal mesenchyme of the cranial nerve ridge , and the bone of the trunk is derived from the endoderm . The large number of clinical and animal experiments show that the bone donor has higher success rate than other parts of bone . The main expression is higher survival rate and lower long - term absorption rate of the graft donor .


These existing clinical , laboratory and biological studies suggest that bone progenitor cells in bone marrow of different parts may have their own biological characteristics ; the cytologic model for bone tissue repair is mainly derived from long bones . Bone marrow stromal cells derived from bone marrow derived from bone marrow are cultured in vitro .


In vitro culture of bone marrow stromal cells derived from bone marrow stromal cells from the first part of the rat ' s jaw and ilium


Objective : To study the biological characteristics of bone marrow cells from the bone marrow of the bone and ilium of the rat , and to provide a cell model .


Methods : Male Sprague - Dawley rats aged from 4 to 6 weeks old were dissected and isolated from bone marrow cells .


Results : After 24 hours of primary cell inoculation , there was a cell adherent on the bottom of the cell , and most of the peripheral blood cells were removed for the first time to remove most of the spindle cells which were scattered in the peripheral blood cells . The cells were removed gradually with the repeated liquid exchange , and the cells were gradually removed and the cells were grown into a typical fibroblast - colony forming unit ( CFU - F ) . After the first passage of the cells about 2 weeks , the cell proliferation ability of the passaged cells in the 6th generation ( ilium ) - 9th generation ( jaw ) was relatively strong , the cell viability was gradually weakened , and the cells appeared different differentiation tendency , and vacuoles and granular substances appeared in the cytoplasm of some cells , and finally died .


Conclusion : The whole bone marrow cell adherent culture method can be used to cultivate the bone marrow cells which meet the experimental requirements .


Comparative study on the in vitro biological characteristics of bone marrow stromal cells derived from bone marrow stromal cells from the second part of the rat ' s jaw and ilium


Objective : To investigate the difference of biological characters of bone and iliac bone in vitro in vitro and to provide a basis for further in vivo study .


Methods : To compare the biological characteristics of bone , bone , mineralization and lipid - forming ability of the second - generation normal - growing jaw and ilium , respectively .


Results : Compared with ilium , bone marrow derived from bone marrow had stronger proliferative ability under low concentration inoculation condition , higher alkaline phosphatase positive expression after induction of bone formation , and formed more calcium nodules after mineralization induction .


Conclusion : The bone and ilium sources have different biological characteristics in vitro , suggesting that they may possess their unique biological characteristics and provide experimental basis and reference for further study of the mechanism of mandibular lesions and regenerative repair .

【學(xué)位授予單位】：南京醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2008
【分類(lèi)號(hào)】：R329

【引證文獻(xiàn)】

相關(guān)期刊論文 前1條

1 鄒薇;劉慧通;李騰;王民;;種子細(xì)胞在頜骨缺損修復(fù)中的研究與應(yīng)用[J];中國(guó)組織工程研究;2012年41期

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本文編號(hào)：1700572