转化生长因子-β3联合牙髓干细胞在种植体骨结合中的作用

发布时间：2018-02-06 05:35

  本文关键词： 牙髓干细胞 转化生长因子-β3 种植体　出处：《新疆医科大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的探讨外源性TGF-β3联合体外培养的DPSCs在种植体骨结合中的作用。方法(1)采用酶解组织块法从新西兰幼兔牙髓组织中分离DPSCs,体外培养,光镜下行细胞形态学观察,将DPSCs传至P3,液氮储存待用;(2)将36只2月龄新西兰大白兔随机分为PBS组;DPSCs组;TGF-β3+DPSCs组,每组12只。在兔下颌前牙及磨牙区随机拔除两颗牙齿,在拔牙窝即刻植入种植体,PBS组植入Bio-Oss粗颗粒骨粉0.30g+PBS 20ul;DPSCs组植入Bio-Oss粗颗粒骨粉0.30g+1×108/L DPSCs 20ul;TGF-β3+DPSCs组植入Bio-Oss粗颗粒0.30g+80ng/L TGF-β3 20ul+1×108/L DPSCs20ul。DPSCs+TGF-β3组则植入1×108/L DPSCs 20ul+80ng/L TGF-β3 20ul。术后第四周、第八周分别处死新西兰大白兔各18只,行HE染色、茜素红染色检测成骨质量,行免疫组织化学法检测骨涎蛋白(bone sialoprotein,BSP)、I型胶原蛋白(collagenase-Ⅰ,COL-Ⅰ)及骨钙素(osteocalcin,OC)的表达,行骨形态计量学分析检测种植体周围骨板宽度(combined bone lamella width,CBLW)、骨结合率(implant bone contact rate,IBCR)、骨小梁宽度(trabecular width,TW)及骨小梁视野面积百分比(trabecular area,TA)。结果原代培养的兔DPSCs呈集落生长,多呈梭形或纺锤形,少数呈多角形;HE染色:术后4周,TGF-β3+DPSCs组在骨小梁数量、成骨细胞密度、新骨形成方面均明显强于其他两组,术后8周,新骨形成进一步增多;茜素红染色:术后4周,TGF-β3+DPSCs组较其他两组出现更多的红色钙化结节,术后8周,钙化结节进一步增多;免疫组织化学图像灰度分析显示:术后4周,TGF-β3+DPSCs组BSP、OC及COL-I指标为(0.35±0.04)、(0.36±0.03)及(0.39±0.01),DPSCs组BSP、OC及COL-I指标为(0.27±0.02)、(0.24±0.01)及(0.28±0.03),PBS组BSP、OC及COL-I指标为(0.13±0.03)、(0.15±0.02)及(0.16±0.02),术后8周,TGF-β3+DPSCs组BSP、OC及COL-I指标为(0.51±0.02)、(0.49±0.03)及(0.53±0.02),DPSCs组BSP、OC及COL-I指标为(0.35±0.02)、(0.37±0.01)及(0.38±0.01),PBS组BSP、OC及COL-I指标为(0.21±0.03)、(0.19±0.02)及(0.22±0.02),术后4周及8周,TGF-β3+DPSCs组BSP、OC及COL-I的表达均显著高于DPSCs组及PBS组(P0.05),DPSCs组及PBS组差异无统计学意义(P0.05)。;术后8周,TGF-β3+DPSCs组CBLW、IBCR、TW、TA均显著高于DPSC组及PBS组(P0.05),DPSCs组及PBS组差异无统计学意义(P0.05)。结论DPSCs具有高度增殖的生物学特性,并具有成骨向分化潜能;TGF-β3对DPSCs成骨向分化具有一定促进作用;TGF-β3联合DPSCs可有效促进种植体的骨结合。
[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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