夏茜,吴镭,毛久凤,郭艺,曾筱,李治,王碧超,董强
XIA Qian,WU Lei,MAO Jiufeng,GUO Yi,ZENG Xiao,LI Zhi,WANG Bichao,DONG Qiang

• 1:贵州医科大学附属口腔医院口腔内科
• 2:贵州医科大学口腔医学院口腔修复学教研室
• 3:贵州医科大学附属省人民医院口腔修复科
• 4:贵州医科大学口腔医学院口腔正畸学教研室

摘要(Abstract)：

目的:研究不同表面钛片上成骨细胞膜片的生长增殖情况。方法:制备光滑表面钛片(对照组)和喷砂酸蚀表面钛片(实验组),把鉴定后的体外培养原代成骨细胞置两种钛片上制备成骨细胞膜片;通过电镜扫描(SEM)观察两组钛片上的成骨细胞形态及附着情况,采用CCK-8法评估两组钛片表面对成骨细胞膜片生长增殖的影响。结果:SEM可见对照组钛片表面较光滑,成骨细胞附着后多单层均匀生长,基质连接处有断裂;实验组钛片表面粗糙,形成大小不一,凹凸不平的"蜂窝状"多级微孔,细胞黏附后多层生长,基质连接紧密;CCK-8结果显示,两组膜片吸光度(OD)值随培养时间延长而增高,实验组各时间点OD值均高于对照组,除第1、3天外,其它时间点OD值比较,差异有统计学意义(P<0.05)。结论:喷砂酸蚀钛表面对成骨细胞膜片增殖促进作用明显高于光滑钛表面。
Objective: To study the growth and proliferation of osteoblast cell sheets on different titanium surfaces. Method: Cultivating smooth surface titanium sheets(control group) and sand acid etched titanium sheets(experiment group); planting the cultivated in vitro osteoblast cell on the two titanium surfaces to be osteoblast cell sheets. Observing the form of different titanium surfaces and the condition of cell attachment through Scanning Electron Microscope(SEM). The effect of growth and proliferation of osteoblast cell sheets on both surfaces were assessed by using Cell Counting Kit 8(CCK-8). Result: Observing by SEM,the surface of control group was smooth and the osteoblast cell sheets growth were monolayer after cell attachment,and there are some fractures of cellular matrix.However,the surface of experimental group was rough and formed alveolate multistage microporous,the osteoblast cell sheets growth were multilayer after cell attachment,cellular matrix connection was tight.CCK-8 indicated that OD value of osteoblast cell sheets of each group increased with culturing time extension. At each time point,the OD value of experimental group is higher than that of control group,except the first day and the third day,other time point has statistical significance(P < 0. 05). Con-clusion: Compared with the surface of smooth titanium,the surface of sand acid etched titanium has a more significant role on the osteoblast cell sheets proliferation.

关键词(KeyWords)： 成骨细胞膜片;光滑钛表面;喷砂酸蚀钛表面;CCK-8
osteoblast cell sheets;smooth titanium surface;sand acid etched titanium surface;CCK-8

Abstract:

Keywords:

基金项目(Foundation): 贵州省科技厅省校合作协议项目[黔科合LH字(2014)7115号];; 贵阳市科技计划项目[筑科合同(2014)100139号]

作者(Author): 夏茜,吴镭,毛久凤,郭艺,曾筱,李治,王碧超,董强
XIA Qian,WU Lei,MAO Jiufeng,GUO Yi,ZENG Xiao,LI Zhi,WANG Bichao,DONG Qiang

DOI: 10.19367/j.cnki.1000-2707.2017.05.009

参考文献(References)：

• [1]Gusic'N,Ivkovic'A,Va Faye J,et al.Nanobiotechnology and bone regeneration:a mini-review[J].Int Orthop,2014(9):1877-1884.
• [2]Ran JB,Hu JX,Sun GL,et al.Comparisons between gelatin-tussah silk fibroin/hydroxyapatite and gelatinBombyxmori silkfibroin/hydroxyapatite nano-composites for bonetissue engineering[J].RSC Advances,2015(93):76526-76537.
• [3]Okano T,Yamada N,Sakai H,et al.A novel recovery system for cultured cells using plasma-treated polystyrene dishes grafted with poly(N-isopropylacrylamide)[J].Biomed Mater Res,1993(10):1243-1251.
• [4]Yang J,Yamato M,Shimizu T,et a1.Reconstruction of functional tissues with cell sheet engineering[J].Biomaterials,2007(34):5033-5043.
• [5]Sakaguchi K,Shimizu T,Okano T,et al.Construction of three-dimensional vascularized cardiac tissue with cell sheet engineering[J].Journal of Controlled Release,2015(1):83-88.
• [6]Amagai Y,Karasawa K,Kyungsook J,et a1.Development of a novel carrier optimized for cell sheet transplantation[J].Biomatter,2015(1):1027-1036.
• [7]Ohashi K,Okano T.Functional tissue engineering of the liver and islets[J].The Anatomical Record,2014(1):73-82.
• [8]Ohki T,Yamato M,Ota M,et al.Application of regenerative medical technology using tissue-engineered cell sheets for endoscopic submucosal dissection of esophageal neoplasms[J].Digestive Endoscopy,2015(2):182-188.
• [9]周成菊,周建奖,杨红,等.抗坏血酸对大鼠成骨细胞膜片BMP-2 mRNA表达的影响[J].实用口腔医学杂志,2015(3):305-308.
• [10]Uchihara Y,Akahane M,Shimizu T,et al.Osteogenic matrix cell sheets facilitate osteogenesis in irradiated rat bone[J].Biomed Res Int,2015(1):1-7.
• [11]Liu X,Zhao X,Li B,et a1.Ultra-violet iradiation generate bioactive nano-Ti O2 surface[J].Applied Physics Letters,2008(4):544-552.
• [12]周屹立,丁仲娟,唐玲.钛片表面粗糙度和氧化膜对成骨细胞增殖和分化的影响[J].上海口腔医学,2013(3):282-286.
• [13]毛久凤,夏茜,吴镭,等.不同表面处理方法对钛表面成骨细胞膜片ALP活性的影响[J].贵州医科大学学报,2016(41):410-413.

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