本文選題：多孔鉭 + RGD多肽��； 參考：《南方醫(yī)科大學(xué)》2016年博士論文

【摘要】：第一章RGD多肽修飾國產(chǎn)多孔鉭支架材料的制備目的：利用環(huán)RGD多肽表面修飾國產(chǎn)多孔鉭支架材料,探討RGD多肽修飾前、后多孔鉭支架材料的形貌特征及親水性變化。方法：1 RGD多肽修飾多孔鉭支架材料制備方法：多孔鉭片經(jīng)消毒后浸泡于濃度為100μmol/L的環(huán)RGD多肽磷酸鹽緩沖液中,室溫下不停振動反應(yīng)24h,PBS沖洗3次,在層流分流室自然干燥,后經(jīng)紫外線照射消毒,備用。2多孔鉭形貌特征觀察：大體及掃描電鏡觀察經(jīng)RGD多肽修飾前、后多孔鉭。3支架材料親水性測定：取RGD多肽修飾前、后的各6枚鉭片分別稱重(W1),于室溫下浸泡于去離子水中24h,濾紙除去多余水分,電子天平準確記錄其重量(W2)。利用支架材料吸水率計算公式(吸水率=[(W2-W1)/W1]×100%),測定兩組樣品吸水率,并行統(tǒng)計學(xué)分析。結(jié)果：1多孔鉭經(jīng)RGD多肽修飾前、后形貌特征變化：大體觀察兩組支架材料外觀顏色均為灰黑色,表面光潔。表面及斷面可見蜂窩狀孔隙,分布均勻。掃面電鏡觀察兩組支架材料表面可見孔徑約400-600μm的微孔結(jié)構(gòu),孔隙內(nèi)部有直徑為50～200μm的連通小孔。其中RGD多肽修飾后支架材料表面可見斑點狀涂層,分布薄厚均勻。2支架材料親水性測定：測得RGD多肽修飾前、后多孔鉭吸水率分別為1.98±0.37%,4.83±0.30%。RGD多肽修飾后多孔鉭支架材料吸水率明顯高于修飾前支架,經(jīng)統(tǒng)計學(xué)分析兩者比較有顯著性差異(P0.01)。結(jié)論：RGD多肽修飾國產(chǎn)多孔鉭支架形貌特征未發(fā)生明顯變化,而其親水性明顯提高。第二章RGD多肽修飾國產(chǎn)多孔鉭支架材料對成骨細胞增殖、黏附影響的體外實驗?zāi)康模和ㄟ^體外實驗研究探討RGD多肽修飾多孔鉭支架材料對成骨細胞增殖、早期黏附等生物學(xué)行為的影響,為國產(chǎn)多孔鉭作為骨組織工程支架材料修復(fù)骨缺損進行體內(nèi)實驗及臨床應(yīng)用提供實驗依據(jù)。方法：1成骨細胞分離培養(yǎng)：取新生24 h內(nèi)新西蘭乳兔4只,無菌條件下取顱蓋骨,剪成1mm×1mm的小塊,置入5ml 0.25%胰蛋白酶離心管中37℃消化30min,加入5ml 0.1％II型膠原酶37℃振蕩消化60min,離心后所得沉淀物加入完全培養(yǎng)基,吹打成細胞懸液,使細胞濃度達105/ml,接種于25cm2培養(yǎng)瓶中。待細胞生長至鋪滿培養(yǎng)瓶底80-90%時,進行傳代,取生長狀態(tài)良好的第2代細胞用于實驗。2細胞形態(tài)學(xué)觀察：倒置相差顯微鏡下觀察細胞生長情況并記錄下各個時期的細胞形態(tài)變化。3成骨細胞鑒定：取生長狀態(tài)良好的第2代細胞,接種于預(yù)置無菌蓋玻片的12孔板內(nèi),當細胞長滿80～90%時,進行ALP染色,鑒定成骨細胞。4實驗分組及成骨細胞在多孔鉭支架材料培養(yǎng),形態(tài)學(xué)及生長情況觀察：實驗分組A組：環(huán)RGD修飾多孔鉭組,B組：單純多孔鉭組。空白對照組：將成骨細胞接種于不放支架的24孔板。將第2代生長良好的成骨細胞與多孔鉭復(fù)合培養(yǎng),經(jīng)倒置相差顯微鏡觀察成骨細胞形態(tài)變化及生長情況。5 RGD多肽修飾多孔鉭對成骨細胞黏附的影響：取生長狀態(tài)良好的第2代成骨細胞,將其調(diào)整成濃度為1×106/ml單細胞懸液,用微量移液器吸取100μ1(1×105個細胞)接種到各組支架表面,于2h、4h分別取出各組支架樣本,利用沉淀法檢測各組支架成骨細胞黏附率,并行統(tǒng)計學(xué)分析。通過掃面電鏡觀察4h時兩組鉭支架表面成骨細胞黏附形態(tài)、數(shù)量情況。6 RGD多肽修飾多孔鉭支架材料對成骨細胞增殖的影響：取濃度為2.5×105/ml的單細胞懸液200μl(5×104個細胞)接種到各組支架表面,培養(yǎng)1d、3d、5d、7d,在每個時間點培養(yǎng)基內(nèi)加入MTT溶液,使用BIO-RAD酶聯(lián)免疫檢測儀(測定波長為490nm,參比波長為650nm)測定各組光吸收值(OD值),繪制各組成骨細胞增殖曲線圖,并對不同時間點各組成骨細胞增殖率進行統(tǒng)計學(xué)分析。通過掃面電鏡觀察3d時兩組鉭支架表面成骨細胞形態(tài)及生長增殖情況。結(jié)果：1成骨細胞分離培養(yǎng)及鑒定：剛接種的原代細胞懸浮于培養(yǎng)基中,倒置相差顯微鏡下可見細胞呈透亮的圓球形,且大小一致；接種6h時,細胞有部分貼壁,24h左右細胞完全貼壁鋪伸。第3d時,細胞數(shù)量增多,體積增大,細胞借偽足樣突起相互連接。第6-7d時細胞數(shù)量繼續(xù)增多,相互融合幾乎鋪滿培養(yǎng)瓶底部,成單層或不規(guī)則樣改變。第2代細胞形態(tài)趨于單一化,呈長梭形或多角形。經(jīng)ALP染色可見細胞漿內(nèi)可見藍染顆粒,細胞核為紅色,證實為成骨細胞。2倒置相差顯微鏡下觀察成骨細胞與多孔鉭支架材料復(fù)合培養(yǎng)形態(tài)特征及生長情況：兩組多孔鉭材料邊緣黏附的細胞逐漸數(shù)量增多,排列密集,形態(tài)良好,但A組細胞數(shù)量、密集程度均高于B組。3成骨細胞黏附率的測定：在2h、4h兩個時間點上,A組黏附率均高于B組及空白對照組,比較差異有統(tǒng)計學(xué)意義(P0.05)；而B組與空白對照組黏附率相近,二者無統(tǒng)計學(xué)差異(P0.05)。各組細胞黏附率在2h、4h兩時間點之間比較均有統(tǒng)計學(xué)差異(P0.05)。4成骨細胞增殖率的測定：第1、3、5、7d四個時間點各組間比較,A組OD值均高于B組及空白對照組,并且差異具有統(tǒng)計學(xué)意義(P0.05);而B組與空白對照組OD值相近,差別無統(tǒng)計學(xué)意義(P0.05)。5掃描電鏡下觀察成骨細胞在支架上的形態(tài)特征及生長情況：在4h時,A組支架材料上成骨細胞伸展變形較好,而B組支架材料上成骨細胞多數(shù)為類圓形,伸展變形較慢,同時A組支架材料表面黏附細胞較密集,數(shù)量明顯高于B組。在第3d時,兩組支架材料上成骨細胞生長狀態(tài)均良好,細胞之間結(jié)合緊密,其中A組多孔鉭表面的細胞密度及細胞分泌胞外基質(zhì)的情況均優(yōu)于B組。結(jié)論：1國產(chǎn)多孔鉭支架材料具有良好的生物相容性,無細胞毒性,對成骨細胞的黏附、增殖沒有影響。2 RGD多肽修飾多孔鉭支架材料對成骨細胞的增殖、黏附有促進作用,有望成為骨組織工程的理想支架。第三章RGD多肽修飾國產(chǎn)多孔鉭支架修復(fù)兔橈骨節(jié)段性骨缺損的實驗研究目的：建立兔橈骨節(jié)段性骨缺損模型,通過影像學(xué)、組織學(xué)、生物力學(xué)等多種檢測方法評價RGD多肽修飾多孔鉭支架體內(nèi)生物相容性及骨缺損的修復(fù)能力,從而為國產(chǎn)多孔鉭臨床應(yīng)用提供體內(nèi)實驗依據(jù)。方法：1材料準備及分組：將多孔鉭材料制成直徑為3.5mm,長15mm的圓柱體,經(jīng)濃度為100μmol/L的RGD多肽溶液修飾后(操作步驟見第一章),紫外線照射消毒,備用。選取105只6-8月齡新西蘭大白兔,隨機抽取分為5組,A組：多孔鉭+RGD多肽組24只,B組：多孔鉭+筋膜包裹組24只,C組：單純多孔鉭組24只,D組：異種骨組24只,E組：空白組9只。2手術(shù)步驟：麻醉成功后右前肢切口周圍備皮,消毒后取右前臂橈側(cè)中段縱形切口,長約3.5cm,以橈骨弧頂為中心,用卡尺測量出15mm長度,用微型擺鋸低速截骨,造成一1.5cm節(jié)段性骨缺損,生理鹽水沖洗傷口后,以嵌插方式分別植入A、B、C、D組植入物。其中B組筋膜瓣制作：沿橈骨中段切開皮膚,游離并切取約30mmx25mm富含毛細血管網(wǎng)的帶蒂筋膜瓣,包裹嵌插至橈骨骨缺損處的植入物。E組為空白組,只做截骨,未加植入物。逐層閉合傷口。術(shù)后給予預(yù)防感染治療。3術(shù)后實驗兔一般情況：觀察動物飲食、日常活動及傷口愈合情況。4X線檢查：分別于術(shù)后當日、4、8、16周行實驗兔右橈骨正位X線檢查,觀察界面骨愈合情況。5大體觀察：術(shù)后4、8、16周三個時間點,切除各組橈骨標本表面軟組織,肉眼觀察骨缺損部位的大體修復(fù)情況。6組織學(xué)檢查：術(shù)后4、8、16周三個時間點,各組標本經(jīng)脫鈣、石蠟切片HE染色及不脫鈣硬組織切片甲苯胺藍染色觀察植入物材料與宿主骨界面及材料內(nèi)部新生骨生長情況。7植入物與骨界面掃描電鏡觀察：上述三個時間點觀察A、B、C、D組界面、材料表面及孔隙內(nèi)部類骨樣組織生長情況。9生物力學(xué)檢測：取術(shù)后16周A、B、C、D組完整橈骨及對側(cè)正常橈骨標本行三點彎曲試驗,檢測修復(fù)后橈骨生物力學(xué)性能。9 Micro-CT掃描評估：取術(shù)后16周A、B、C、D組以植入物為中心,截取長度為2.5cm帶尺、橈骨標本行Micro-CT掃描、三維重建及新生骨計量學(xué)分析。結(jié)果：1術(shù)后動物一般情況：術(shù)后3-7天后動物飲食、精神逐漸恢復(fù)正常,切口部位無紅腫、滲液及化膿等,均為I期愈合；2周后肢體活動基本恢復(fù)正常,跛行消失,體重恢復(fù)到術(shù)前水平。2 X線觀察：術(shù)后當日X線可見各組植入物位置良好,無明顯位移。術(shù)后4、8、16周三個時間點可見隨時間延長各組植入物與宿主骨界面結(jié)合越來越牢固,骨折線逐漸消失,其中D組骨痂塑形良好,骨髓腔部分再通；在A、B、C三組中,A組界面骨痂生成最多,塑形良好；E組術(shù)后16周仍可見骨缺損,兩斷端骨質(zhì)逐漸硬化,髓腔封閉。3大體標本觀察：術(shù)后4、8、16周三個時間點A組材料表面逐漸被骨樣組織包裹,界面與宿主骨結(jié)合牢固,塑形良好。B、C組界面結(jié)合牢固,表面材料孔隙大部分可見骨樣組織填充,靠近界面部分表面被骨組織覆蓋。D組植入材料逐漸降解,材料降解部分被新生骨樣組織替代,有明顯的毛細血管懌長入,材料與宿主骨界面融合良好、牢固。術(shù)后4周空白組斷端可見少量骨痂生長,16周時斷端骨質(zhì)硬化,光滑,髓腔已經(jīng)封閉,缺損處無骨性連接形成。4組織學(xué)檢查：術(shù)后4、8、16周各組標本經(jīng)脫鈣、石蠟切片HE染色及不脫鈣硬組織切片甲苯胺藍染色,光鏡觀察可見隨著時間推移,A-C三組鉭-宿主骨界面新生骨組織數(shù)量逐漸增多并沿多孔鉭孔隙長入材料內(nèi)部,骨組織由幼稚到成熟；D組異種骨逐漸降解并被新生骨組織包裹替代；而E組(空白對照組)骨缺損始終存在,洞壁被薄層纖維膜內(nèi)襯。5植入物與骨界面掃描電鏡觀察：術(shù)后4、8、16周三個時間點觀察可見A、B、C三組鉭-骨界面、材料表面及孔隙內(nèi)部骨膠原及成骨細胞逐漸增多,新生骨逐漸成熟并過度為板層骨,鉭-骨界面縫隙逐漸結(jié)合緊密；D組新生骨組織逐漸增多,并分化為成熟狀小梁骨。6生物力學(xué)檢測：術(shù)后16周對A、B、C、D組及對照組正常完整橈骨行三點彎曲試驗測定最大載荷力及抗彎曲強度,結(jié)果顯示A、B、C、D四組無論是最大載荷力還是抗彎曲強度均低于正常橈骨組,并且有統(tǒng)計學(xué)差異(P0.01)。A、B、C、D四組間比較,A組力學(xué)性能最高,B、C組次之,D組最低,經(jīng)單因素方差分析各組間差異均有統(tǒng)計學(xué)意義(P0.01)。7 Micro-CT掃描觀察：術(shù)后16周,A、B、C、D四組植入物界面、表面均有大量骨組織覆蓋,內(nèi)部不同程度新生骨組織填充。新生骨體積分數(shù)定量分析結(jié)果顯示D組新生骨組織百分比高于A、B、C三組,其中A組與D組無統(tǒng)計學(xué)差異(P0.05),B、C組與D組有統(tǒng)計學(xué)差異(P0.01); A, B, C三組間有統(tǒng)計學(xué)差異(P0.01)。結(jié)論：國產(chǎn)多孔鉭材料具有良好的生物相容性,RGD多肽修飾多孔鉭支架材料骨傳導(dǎo)能力更強,修復(fù)兔橈骨節(jié)段性骨缺損效果肯定。
[Abstract]:In the first chapter, the preparation of domestic porous tantalum scaffold materials with RGD polypeptide modified homemade porous tantalum scaffold materials: the surface modification of domestic porous tantalum scaffold materials on the surface of RGD polypeptide was used to explore the morphologies and hydrophilicity of the porous tantalum scaffold material before the modification of RGD polypeptide. Method: the preparation method of porous tantalum scaffold materials with 1 RGD polypeptide modified porous tantalum scaffold: porous tantalum slices were dipped after disinfection In the ring RGD polypeptide phosphate buffer solution with a concentration of 100 mol/L, the vibrational reaction of 24h, PBS flush for 3 times at room temperature, then naturally dried in the laminar flow division chamber, and then sterilized by ultraviolet radiation, and observed by the.2 porous tantalum morphology: the hydrophilicity of the porous tantalum.3 scaffold was measured by the general and scanning electron microscope before the RGD polypeptide was modified. Before the modification of RGD polypeptide, the 6 tantalum slices were weighed (W1) respectively. At room temperature, they were soaked in the deionized water 24h, the filter paper removed the excess water, and the weight was recorded accurately by the electronic balance (W2). The water absorption rate of the scaffold was calculated by using the water absorption rate = [(W2-W1) /W1] x 100%), and the water absorption rate of the two groups was measured, and the results were statistically analyzed. The results were 1 porous. The morphology of tantalum was changed after RGD peptide modification. The appearance of two groups of scaffold materials were all gray and black, and the surface and surface were smooth. The surface and cross section showed the honeycomb pore and the distribution was uniform. The surface and the surface of the two groups of scaffolds were observed to have a pore structure of about 400-600 mu m, with a diameter of 50~200 mu m in the pores. On the surface of RGD polypeptide, the surface of the scaffold material can be seen on the surface of the scaffold. The hydrophilicity of the thin and uniform.2 scaffold material is measured. The water absorption rate of the porous tantalum was 1.98 + 0.37% before the RGD polypeptide modification, and the water absorption of the porous tantalum scaffold was significantly higher than that of the pre modified scaffold after the 4.83 + 0.30%.RGD polypeptides modified. Two There was a significant difference (P0.01). Conclusion: the morphology of the domestic porous tantalum scaffold modified by RGD polypeptide did not change obviously, but its hydrophilicity was obviously improved. The effect of the second chapter RGD polypeptide modified domestic porous tantalum scaffold material on the proliferation and adhesion of osteoblasts was in vitro: the modification of RGD polypeptide through the experiment in vitro The effects of porous tantalum scaffold on the biological behavior of osteoblast proliferation and early adhesion are provided for the experiment and clinical application of domestic porous tantalum as scaffold material for bone tissue engineering to repair bone defects. Methods: 1 osteoblasts were isolated and cultured: 4 New Zealand New Zealand milk rabbits were taken from 24 h, and the skull was taken under aseptic conditions. The small pieces of 1mm x 1mm were cut into the 5ml 0.25% trypsin centrifuge tube to digest 30min at 37 degrees C, and 5ml 0.1%II collagenase was added to the 60min. The precipitates were added to the complete medium after centrifugation, and the cells were blown into cell suspension, and the cell concentration was 105/ml and inoculated into the 25cm2 culture bottle. When the cell was grown to the bottom 80-90% of the culture bottle, The second generation cells with good growth state were used to observe the morphological observation of the experimental.2 cells: the cell growth was observed under the inverted phase contrast microscope and the morphological changes of the cells at various stages were recorded and.3 osteoblasts were recorded. The second generation cells with good growth state were inoculated to the 12 foramen of the pre sterile cover glass, and the cell length was long. At the time of 80 to 90%, ALP staining was carried out to identify the.4 experimental group of osteoblasts and the culture of osteoblasts in the porous tantalum scaffold materials, morphology and growth. The experimental group A group: ring RGD modified porous tantalum group, B group: pure porous tantalum group. The blank control group was inoculated to the 24 hole plate without stent. The second generation good growth was good. Good osteoblasts were cultured with porous tantalum, and the morphology and growth of osteoblasts were observed by inverted phase contrast microscope. The effect of.5 RGD polypeptide on the adhesion of porous tantalum on osteoblast: second generation of osteoblasts with good growth status were adjusted to a concentration of 1 x 106/ml single cell suspension, and 100 micron 1 (1 x) was absorbed by a micropipette. 105 cells were inoculated on the surface of each scaffold. The scaffold samples were taken out by 2H and 4H respectively. The adhesion rate of osteoblasts was detected by precipitation method. The adhesion morphology of the two groups of tantalum scaffolds on the surface of tantalum scaffold was observed by scanning electron microscopy. The number of.6 RGD polypeptide modified the porous tantalum scaffold material to osteoblasts. The effect of proliferation: a single cell suspension of 2.5 * 105/ml (5 x 104 cells) was inoculated to the surface of each scaffold, and 1D, 3D, 5D, 7d were cultured. MTT solution was added to each time point culture, and the BIO-RAD enzyme immunodetector (wavelength 490nm, reference wavelength was 650nm) was used to determine the optical absorption values of each group (OD value), and the composition of each component was plotted. The proliferation rate of bone cells in different time points was statistically analyzed. The morphology and growth and proliferation of two groups of tantalum scaffolds on the surface of tantalum scaffold were observed by scanning electron microscopy. Results: 1 osteoblasts were isolated and cultured and identified: primary cell suspension in the culture medium and inverted phase contrast microscope in the two groups At the time of inoculation 6h, the cells were partially adhered to the cells, and the cells were partially adhered to the wall, and the cells were completely plaster and extended. At the time of 3D, the number of cells increased, the volume increased, and the cells were connected by the pseudo foot like protrusions. At the time of 6-7d, the number of cells continued to increase and the phase interfusion almost covered the bottom of the culture bottle, formed monolayer or irregularity. The morphology of the second generation of cells tended to be single, with long spindle shape or polygon. The blue dye particles were visible in the cytoplasm by ALP staining, and the nuclei were red. It was confirmed that the morphological characteristics and growth conditions of osteoblasts and porous tantalum scaffold materials were observed under the inverted phase contrast microscope of.2 osteoblasts: the marginal viscosity of the two groups of porous tantalum materials. The number of cells in the attached cells was increased and the morphology was good, but the number of cells in the A group was higher than that of the B group.3 osteoblast adhesion rate. At the two time points of 2H and 4h, the adhesion rate of group A was higher than that of the B group and the blank control group, and the difference was statistically significant (P0.05), but the adhesion rate of the B group and the blank control group was similar, and the rate of adhesion was similar in the B group and the blank control group. There was no one in the group of B and the blank control group. Statistical difference (P0.05). The cell adhesion rate of each group at 2h, 4H two time points were statistically different (P0.05) the proliferation rate of.4 osteoblast: the 1,3,5,7d four time points were compared in each group, the A group was higher than the B group and the blank control group, and the difference was statistically significant (P0.05), while the B group was similar to the blank control group. There was no statistical significance (P0.05).5 scanning electron microscope to observe the morphological characteristics and growth of osteoblasts on the scaffold: at 4h, the osteoblasts in the A stents were extended and deformed, while the majority of the osteoblasts in the B group were round, and the extensional deformation was slow, and the surface adhesion cells of the A stents were denser and more dense. It was significantly higher than group B. At 3D, the growth state of osteoblasts on the two groups of scaffolds was good and the cells were tightly bonded. The cell density on the surface of the porous tantalum on the A group and the secretion of extracellular matrix were superior to that of the B group. Conclusion: 1 domestic porous tantalum scaffold materials have good biocompatibility, no cytotoxicity, and osteoblasts. Adhesion, proliferation does not affect the proliferation of.2 RGD polypeptide modified porous tantalum scaffold materials for osteoblasts. Adhesion has a promoting effect, and it is expected to be an ideal scaffold for bone tissue engineering. Third experimental research on the repair of rabbit radial segmental bone defect with homemade porous tantalum scaffold modified by RGD polypeptide: to establish a rabbit radial segmental bone defect model, The biocompatibility and repair ability of RGD polypeptides modified porous tantalum scaffold were evaluated by various methods of imaging, histology and biomechanics, so as to provide the experimental basis for the clinical application of domestic porous tantalum. Method: 1 material preparation and grouping: the porous tantalum material was made into a cylinder with a diameter of 3.5mm and a long 15mm. After the modification of the RGD polypeptide solution with a concentration of 100 mol/L (see Chapter 1), UV irradiation and disinfection, the 105 6-8 month old New Zealand white rabbits were selected randomly and divided into 5 groups randomly, A group: 24 porous tantalum +RGD polypeptide group, 24 B group: porous tantalum + fascia package group, C group: pure porous tantalum group 24, D group, 24 bone group, E Group: 9 steps of.2 operation in the blank group: skin preparation around the right forelimb incision after anesthesia was successful. After disinfection, the right forearm radial middle section was taken longitudinally. The length of the radial incision was about 3.5cm. The length of 15mm was measured with a caliper. A 1.5cm segmental bone defect was caused by the micro sawing the low speed osteotomy. After the saline was used, the wound was inserted into the wound. Do not implant the implants of group A, B, C, and D. Among them, group B was made of fascial flap: incision of the skin along the middle of the radius, free and cut off the pedicled fascial flap with the capillary network of 30mmx25mm, and the.E group inserted into the radius bone defect in group.E as the blank group, only osteotomy, no implants and closed wounds. Postoperative.3 surgery was given to prevent infection for.3 surgery. The general condition of the rabbit after the experiment: Observation of animal diet, daily activity and wound healing.4 X - ray examination: on the day after the operation, the right radiography of the right radius of the rabbit was examined on the day of 4,8,16, and the bone healing of the interface was observed by the gross observation of.5: the surface soft tissue of the radius specimens were removed and the naked eye was observed by the naked eye after the operation of the 4,8,16 Wednesday. General repair of the site.6 histological examination: 4,8,16 Wednesday postoperatively, specimens of each group were decalcified, paraffin section HE staining, and DT staining was used to observe the interface between the implant and the host bone and the growth of the internal bone in the material. The.7 implants and the bone interface were observed by scanning electron microscope: the three time A, B, C, D group interface, material surface and internal bone like tissue growth of.9: 16 weeks after the operation: A, B, C, D group complete radius and three point bending test on the contralateral normal radius, and detect the.9 Micro-CT scanning of the radial biomechanics after repair: 16 weeks after the operation. Heart, the length of intercepted length was 2.5cm tape, Micro-CT scan of radial mark, three-dimensional reconstruction and new bone Metrology Analysis. Results: 1 animal general condition after operation: 3-7 days after operation, the animal diet, the spirit gradually resumed normal, the incision site was not red, the seepage and suppuration were all I, after 2 weeks, the extremities were basically restored to normal, limping. .2 X - ray observation of loss, weight recovery to preoperative level: on the day after operation, the X - ray showed that the implants were in good position and no obvious displacement. After 4,8,16 Wednesday, the interface of the implant and the host bone became more and more firm, and the fracture line gradually disappeared, in which the callus of group D was well shaped and the bone marrow cavity was repassed; in A, B, C three groups, A group interface callus formation most, shape good shape, E group 16 weeks after the bone defect still visible, the two broken end bone gradually hardened, the pulp cavity closed.3 gross specimen observation: 4,8,16 Wednesday time point A group material surface gradually wrapped by bone like tissue, the interface and host bone solid, good plastic.B, C group interface firm, C group interface firm, C group.B, C group interface firmly, C Most of the pores in the surface of the surface were filled with bone like tissue, and the.D group was gradually degraded on the surface near the surface of the interface, and the degradation part of the material was replaced by the new bone like tissue. There was a clear capillary vein, and the fusion of the material and the host bone was good and solid. A small amount of callus growth was seen in the 4 weeks after the operation, 1 At 6 weeks, bone sclerosis, smooth, pulp cavity had been closed, and no bone connection was formed in the defect to form a.4 histological examination. After 4,8,16 weeks, the specimens were decalcified, paraffin section HE staining, and decalcified hard tissue section toluidine blue staining. The number of new bone tissue in the tantalum host bone interface of the three groups was observed gradually as time went on, and the number of new bone tissue in the tantalum host bone interface was gradually increased as time went on. The bone tissue was increased from naive to maturity along the porous tantalum pores, and the bone tissue was degraded gradually and replaced by the new bone tissue in D group, while the bone defect existed in group E (blank control group), and the wall was scanned by.5 implants and bone interface by thin layer fibrous membrane, and the observation was observed at the time of 4,8,16 Wednesday after the operation. A, B, C three groups of tantalum bone interface, the material surface and pore bone collagen and osteoblasts increased gradually, the new bone gradually matured and overworked.

【學(xué)位授予單位】：南方醫(yī)科大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2016
【分類號】：R318.08;R687

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