發(fā)布時(shí)間：2018-01-26 22:54

  本文關(guān)鍵詞： 支架材料 雞蛋殼 鋅涂層 臨界性骨缺損 原位組織工程　出處：《復(fù)旦大學(xué)》2012年碩士論文　論文類型：學(xué)位論文

【摘要】：目的 本實(shí)驗(yàn)旨在通過觀察雞蛋殼復(fù)合抗菌支架材料對(duì)兔顱骨臨界性骨缺損的修復(fù)情況,初步研究其修復(fù)骨缺損的能力,探討雞蛋殼復(fù)合抗菌支架材料應(yīng)用于骨原位組織工程中的可行性。 方法 1、雞蛋殼經(jīng)清潔處理后,在研缽內(nèi)研磨成粉,篩子過濾300-500μm。使用微波爐法對(duì)處理過的雞蛋殼進(jìn)行表面改良和鋅涂層。對(duì)表面改良和鋅涂層的雞蛋殼分別進(jìn)行掃描電鏡(SEM)、能量散色光譜儀(EDXA)、X射線衍射儀(XRD)及傅里葉紅外(FT-IR)等理化性能的檢測(cè)。 2、動(dòng)物缺損模型的建立：選用30只雄性新西蘭大白兔,全麻下每只兔顱骨制備5個(gè)6mm直徑硬腦膜全厚層缺損區(qū)。分為A組(空白對(duì)照組,Control組)；B組(雞蛋殼粉末組,ES組)；C組(微波爐法60min表面改良的雞蛋殼粉末組,MES組)；D組(鋅涂層雞蛋殼粉末組,ZnES組);E組(Bio-oss組)。將造模后的家兔隨機(jī)分成三組,術(shù)后4、8、12周分批處死一組實(shí)驗(yàn)動(dòng)物,處死動(dòng)物前用鈣黃綠素和四環(huán)素進(jìn)行熒光標(biāo)記。 3、骨計(jì)量學(xué)標(biāo)本的制作：用低速鋸分離取下整個(gè)顱骨,沿缺損邊緣外圍處切下帶有材料的骨組織。經(jīng)過固定、脫水、滲透、聚合后使用硬組織切片機(jī)沿顱骨缺損模型縱向切成8μm和20μm切片,8μm薄片Goldner-Trichrome三色法染色、20μm厚片在熒光顯微鏡下直接觀察骨組織的細(xì)微病理改變,用圖像分析軟件進(jìn)行參數(shù)測(cè)量、計(jì)算和分析。 4、所得數(shù)據(jù)采用“均值±標(biāo)準(zhǔn)差”(x±S)表示,應(yīng)用SPSS16.0軟件進(jìn)行統(tǒng)計(jì)分析。 結(jié)果 1、蛋殼本身具有類似骨組織的孔隙結(jié)構(gòu),微波加熱技術(shù)可以合成碳磷灰石(CHA)以及復(fù)合了鋅離子的雞蛋殼復(fù)合抗菌支架材料。 2、實(shí)驗(yàn)動(dòng)物術(shù)后生長(zhǎng)恢復(fù)良好,無感染和繼發(fā)性骨折。大體標(biāo)本觀察：空白對(duì)照組缺損腔內(nèi)充滿肉芽組織,ES組、MES組、ZnES組、Bio-oss組材料穩(wěn)定無移位,無脫落,表面被纖維軟骨樣組織所包裹,與自體骨組織緊密牢固連接。 3、熒光雙標(biāo)觀察：空白對(duì)照組僅在缺損邊緣區(qū)域有黃、綠色熒光標(biāo)記帶,ES、MES、ZnES、Bio-oss四組材料不僅缺損邊緣,缺損區(qū)域內(nèi)同樣存在熒光標(biāo)記。對(duì)五組材料缺損邊緣自體骨礦化沉積率進(jìn)行測(cè)定比較,術(shù)后4周、8周、12周各組間差異均無統(tǒng)計(jì)學(xué)意義。說明是否使用骨移植材料以及使用何種骨移植材料對(duì)骨缺損邊緣自體骨的生長(zhǎng)不會(huì)產(chǎn)生影響。對(duì)ES、MES、ZnES、Bio-oss四組材料缺損區(qū)域內(nèi)骨礦化沉積率測(cè)定比較,4周時(shí)各組礦化沉積率差異無統(tǒng)計(jì)學(xué)意義,8周(P0.05)和12周(P0.05)各組差異有統(tǒng)計(jì)學(xué)意義。進(jìn)一步對(duì)8周和12周組間進(jìn)行兩兩比較并得出結(jié)論：ES組礦化沉積率低于MES、ZnES、Bio-oss組,而MES、ZnES、Bio-oss組礦化沉積率組間無統(tǒng)計(jì)學(xué)差異。 4、Goldner-Trichrome染色法：測(cè)量計(jì)算缺損區(qū)域內(nèi)新生骨相對(duì)面積、剩余材料相對(duì)面積并進(jìn)行統(tǒng)計(jì)學(xué)分析。對(duì)同一種材料不同時(shí)間段的缺損區(qū)域內(nèi)新生骨相對(duì)面積進(jìn)行比較：空白對(duì)照組無統(tǒng)計(jì)學(xué)意義,另四組均有統(tǒng)計(jì)學(xué)意義(P0.05)。表明隨著時(shí)間的延長(zhǎng),ES、MES、ZnES、Bio-oss四組材料在缺損區(qū)域內(nèi)的新生骨呈增長(zhǎng)趨勢(shì)。對(duì)同一時(shí)間不同材料缺損區(qū)域內(nèi)新生骨相對(duì)面積進(jìn)行比較：4周、8周、12周不同材料缺損區(qū)新生骨相對(duì)面積差異均有統(tǒng)計(jì)學(xué)意義(P0.05)。進(jìn)一步對(duì)各周組間進(jìn)行兩兩比較并得出結(jié)論：空白對(duì)照組缺損區(qū)域內(nèi)新生骨相對(duì)面積低于其他四組。對(duì)同一材料按不同時(shí)間段的剩余材料相對(duì)面積進(jìn)行比較：ES、MES、ZnES組有統(tǒng)計(jì)學(xué)意義(P0.05),Bio-oss組無統(tǒng)計(jì)學(xué)意義。說明ES、MES、ZnES組材料隨著時(shí)間的延長(zhǎng)呈降解趨勢(shì),而Bio-oss組在實(shí)驗(yàn)期內(nèi)降解不明顯。對(duì)同一時(shí)間不同材料剩余材料相對(duì)面積進(jìn)行比較：4周差異無統(tǒng)計(jì)學(xué)意義,8周和12周差異有統(tǒng)計(jì)學(xué)意義(P0.05)。進(jìn)一步對(duì)8周和12周各組材料剩余材料相對(duì)面積進(jìn)行兩兩比較并得出結(jié)論：ES組降解率優(yōu)于Bio-oss組,但差于MES組和ZnES組。 結(jié)論 雞蛋殼本身具有良好的生物相容性,并可以促進(jìn)新骨再生；表面改良后雞蛋殼的降解性發(fā)生了變化,骨引導(dǎo)效果更好。雞蛋殼復(fù)合抗菌支架材料應(yīng)用于骨原位組織工程修復(fù)骨缺損具有一定的可行性。
[Abstract]:Purpose The purpose of this experiment is to investigate the repair of the critical bone defect of rabbit skull by observing the composite antibacterial scaffold material of egg shell and to investigate the feasibility of applying the composite antibacterial scaffold material in bone in - situ tissue engineering . method 1 . After the egg shell was cleaned , the egg shell was ground into powder , the sieve was filtered 300 - 500 渭m . The treated egg shell was surface modified and zinc coated by microwave oven . Scanning electron microscope ( SEM ) , energy dispersive spectrometer ( EDXA ) , X - ray diffraction ( XRD ) and Fourier infrared ( FT - IR ) were used to detect the surface modification and zinc coating respectively . 2 . Establishment of animal defect model : 30 male New Zealand white rabbits were selected and 5 6mm diameter hard - meninges full thickness defect areas were prepared from each rabbit skull . Group A ( control group ) , group B ( egg shell powder group , MES group ) , group D ( zinc coated eggshell powder group , ZnES group ) , group E ( Bio - oss group ) . 3 . Preparation of bone - metrology specimens : The whole skull was removed by low - speed saw , and the bone tissue with material was cut along the periphery of the defect . After fixation , dehydration , infiltration and polymerization , the bone tissue was cut into 8 渭m and 20 渭m slice along the skull defect model . 4 . The obtained data is expressed by means of " mean 鹵 standard deviation " ( x 鹵 S ) , and SPSS 16.0 software is applied for statistical analysis . Results 1 . The eggshell itself has a pore structure similar to that of bone tissue , and the microwave heating technique can synthesize the carbon apatite ( CHA ) and the composite antibacterial scaffold material of the eggshell of the composite zinc ion . 2 . The growth of experimental animals recovered well , no infection and secondary fracture . Gross specimen observation : The blank control group was filled with granulation tissue , ES group , MES group , ZnES group and Bio - oss group . No significant difference was found between 8 weeks ( P0.05 ) and 12 weeks ( P0.05 ) . The results showed that the rate of mineralization and deposition of ES , MES , ZnES and Bio - oss was lower than that in MES , ZnES and Bio - oss groups , but there was no statistical difference between the groups of MES , ZnES and Bio - oss . 4 銆，

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