發(fā)布時(shí)間：2018-02-06 05:35

  本文關(guān)鍵詞： 牙髓干細(xì)胞 轉(zhuǎn)化生長(zhǎng)因子-β3 種植體　出處：《新疆醫(yī)科大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：目的探討外源性TGF-β3聯(lián)合體外培養(yǎng)的DPSCs在種植體骨結(jié)合中的作用。方法(1)采用酶解組織塊法從新西蘭幼兔牙髓組織中分離DPSCs,體外培養(yǎng),光鏡下行細(xì)胞形態(tài)學(xué)觀察,將DPSCs傳至P3,液氮儲(chǔ)存待用;(2)將36只2月齡新西蘭大白兔隨機(jī)分為PBS組;DPSCs組;TGF-β3+DPSCs組,每組12只。在兔下頜前牙及磨牙區(qū)隨機(jī)拔除兩顆牙齒,在拔牙窩即刻植入種植體,PBS組植入Bio-Oss粗顆粒骨粉0.30g+PBS 20ul;DPSCs組植入Bio-Oss粗顆粒骨粉0.30g+1×108/L DPSCs 20ul;TGF-β3+DPSCs組植入Bio-Oss粗顆粒0.30g+80ng/L TGF-β3 20ul+1×108/L DPSCs20ul。DPSCs+TGF-β3組則植入1×108/L DPSCs 20ul+80ng/L TGF-β3 20ul。術(shù)后第四周、第八周分別處死新西蘭大白兔各18只,行HE染色、茜素紅染色檢測(cè)成骨質(zhì)量,行免疫組織化學(xué)法檢測(cè)骨涎蛋白(bone sialoprotein,BSP)、I型膠原蛋白(collagenase-Ⅰ,COL-Ⅰ)及骨鈣素(osteocalcin,OC)的表達(dá),行骨形態(tài)計(jì)量學(xué)分析檢測(cè)種植體周圍骨板寬度(combined bone lamella width,CBLW)、骨結(jié)合率(implant bone contact rate,IBCR)、骨小梁寬度(trabecular width,TW)及骨小梁視野面積百分比(trabecular area,TA)。結(jié)果原代培養(yǎng)的兔DPSCs呈集落生長(zhǎng),多呈梭形或紡錘形,少數(shù)呈多角形;HE染色:術(shù)后4周,TGF-β3+DPSCs組在骨小梁數(shù)量、成骨細(xì)胞密度、新骨形成方面均明顯強(qiáng)于其他兩組,術(shù)后8周,新骨形成進(jìn)一步增多;茜素紅染色:術(shù)后4周,TGF-β3+DPSCs組較其他兩組出現(xiàn)更多的紅色鈣化結(jié)節(jié),術(shù)后8周,鈣化結(jié)節(jié)進(jìn)一步增多;免疫組織化學(xué)圖像灰度分析顯示:術(shù)后4周,TGF-β3+DPSCs組BSP、OC及COL-I指標(biāo)為(0.35±0.04)、(0.36±0.03)及(0.39±0.01),DPSCs組BSP、OC及COL-I指標(biāo)為(0.27±0.02)、(0.24±0.01)及(0.28±0.03),PBS組BSP、OC及COL-I指標(biāo)為(0.13±0.03)、(0.15±0.02)及(0.16±0.02),術(shù)后8周,TGF-β3+DPSCs組BSP、OC及COL-I指標(biāo)為(0.51±0.02)、(0.49±0.03)及(0.53±0.02),DPSCs組BSP、OC及COL-I指標(biāo)為(0.35±0.02)、(0.37±0.01)及(0.38±0.01),PBS組BSP、OC及COL-I指標(biāo)為(0.21±0.03)、(0.19±0.02)及(0.22±0.02),術(shù)后4周及8周,TGF-β3+DPSCs組BSP、OC及COL-I的表達(dá)均顯著高于DPSCs組及PBS組(P0.05),DPSCs組及PBS組差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05)。;術(shù)后8周,TGF-β3+DPSCs組CBLW、IBCR、TW、TA均顯著高于DPSC組及PBS組(P0.05),DPSCs組及PBS組差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05)。結(jié)論DPSCs具有高度增殖的生物學(xué)特性,并具有成骨向分化潛能;TGF-β3對(duì)DPSCs成骨向分化具有一定促進(jìn)作用;TGF-β3聯(lián)合DPSCs可有效促進(jìn)種植體的骨結(jié)合。
[Abstract]:Objective to investigate the role of DPSCs co-cultured with exogenous TGF- 尾 3 in implant bone binding. Methods DPSCs was isolated from pulp tissue of New Zealand rabbits by enzymolysis tissue block method. In vitro culture, cell morphology was observed under light microscope, DPSCs was transferred to P3, and liquid nitrogen was stored for use. (2) Thirty-six 2-month-old New Zealand white rabbits were randomly divided into PBS group. DPSCs group; TGF- 尾 3 DPSCs group (n = 12) randomly extracted two teeth in the mandibular anterior teeth and molar region and implanted implants in the extraction fossa. In PBS group, 0.30g PBS 20ulwere implanted with Bio-Oss coarse granule bone powder. In DPSCs group, 0.30g / L DPSCs 20ulL of Bio-Oss coarse granulated bone powder was implanted into the bone powder of 0.30g / L 脳 10 ~ (8) / L; TGF- 尾 3 DPSCs group implanted Bio-Oss coarse particles 0.30g 80 ng / L TGF- 尾 3 20ul 1 脳 10 8 / L. DPSCs20ul.DPSCs TGF- 尾 3 group was implanted with 1 脳 10 8% L DPSCs20ul 80ng / L TGF- 尾 3 20ul. At 8th weeks, 18 New Zealand white rabbits were killed, respectively, for HE staining, alizarin red staining for osteogenic quality, and immunohistochemical method for bone sialoprotein sialoprotein. Expression of collagenase- 鈪，

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