摘要
目的探讨不同管径二氧化钛纳米管对人牙龈成纤维细胞(human gingival fibroblast,HGF)生物学行为的影响,以筛选具有良好软组织封闭效果的二氧化钛纳米管。方法在10、30和60 V电压下,利用阳极氧化法在钛表面制备管径分别为30、100和200 nm的二氧化钛纳米管,分别计为30、100和200 nm纳米管组,以未经处理的光滑纯钛片作为对照组。在各组试件表面培养HGF,免疫荧光染色观察2 d后各组细胞黏附形态,甲基噻唑基四唑法检测2 h后细胞黏附情况及培养1、3和7 d时细胞增殖情况,酶联免疫吸附测定检测各组培养7 d后的Ⅰ型胶原分泌量(每组每种实验各3个试件)。结果对照组HGF蜷缩成圆形,30和100 nm纳米管组HGF均呈现明显的同向伸展,200 nm纳米管组HGF分布稀疏且没有伸展。30和100 nm纳米管组细胞黏附A值(分别为0.603±0.021和0.773±0.045)及Ⅰ型胶原分泌量[分别为(36.5±9.5)和(47.7±4.5)μg/ml]均显著大于对照组[细胞黏附A值:0.427±0.057;Ⅰ型胶原分泌量:(22.2±5.9)μg/ml](P<0.05),且100 nm纳米管组细胞黏附A值显著大于30 nm纳米管组(P<0.05);200 nm纳米管组细胞黏附A值(0.250±0.046)及Ⅰ型胶原分泌量[(10.1±3.7)μg/ml]均显著小于对照组(P<0.05)。各时间点100 nm纳米管组细胞增殖A值均为最大,均显著大于相同时间点200 nm纳米管组和对照组(P<0.05),且培养3 d时亦显著大于30 nm纳米管组(P<0.05);各时间点200 nm纳米管组细胞增殖A值均为最小,除培养1 d时与对照组差异无统计学意义(P>0.05)外,均显著小于相同时间点其他组(P<0.05)。结论钛表面管径为100 nm的二氧化钛纳米管较适合HGF的黏附、增殖和Ⅰ型胶原分泌。
Objective To study the effects of titania nanotubes with three different diameters on human gingival fibroblast(HGF).Methods Three groups of specimens were prepared.Titania nanotubes with diameters of 30,100,and 200 nm were synthesized on titanium surfaces through electrochemical anodization at 10,30,and 60 V,respectively.Specimens were assigned into the three groups according to the diameter of the titania nanotubes.Pure smooth titanium without any treatment was set as the control group.HGF were seeded on the surface of the samples.The cell morphology on the specimens was observed with immunofluorescence staining after 2 h,the cell adhesion after 2 d and cell proliferation after 1,3,and 7 d were detected using methyl thiazolyl tetrazolium assay,and the secretion of typeⅠcollagen after 7 d was determined using enzyme-linked immunosorbent assay(each group has three samples for each experiment).Results HGF on the control group exhibited an oval shape without noticeable extensions.HGF on titania nanotubes with a diameter of 30 nm and titania nanotubes with a diameter of 100 nm elongated further and were arranged orderly.HGF on titania nanotubes with a diameter of 200 nm were sparsely distributed without noticeable extensions.Titania nanotubes with a diameter of 30 nm and titania nanotubes with a diameter of 100 nm could enhance the cell attachment(0.603±0.021 and 0.773±0.045),and secretion of typeⅠcollagen[(36.5±9.5)and(47.7±4.5)μg/ml,respectively]compared with the control group whose cell attactment was 0.427±0.057,and secretion of typeⅠcollagen was(22.2±5.9)μg/ml(P<0.05).Furthermore,titania nanotubes with a diameter of 100 nm showed more cell attchment than titania nanotubes with a diameter of 30 nm did(P<0.05).Ttania nanotubes with a diameter of 200 nm clearly impaired the cell adhesion(0.250±0.046)and secretion of typeⅠcollagen[(10.1±3.7)μg/ml]compared with the control group(P<0.05).At each time point,titania nanotubes with a diameter of 100 nm showed the highest cell proliferation,and the amount of cell proliferation was significantly higher than that on the titania nanotubes with a diameter of 200 nm and the control group at each time point(P<0.05),and was also significantly higher than that on the titania nanotubes with a diameter of 30 nm at day three(P<0.05).At each time point,titania nanotubes with a diameter of 200 nm showed the lowest cell proliferation,which was significantly lower than that on the control group at each time point(P<0.05),except that there was no significant difference in the amount of cell proliferation between titania nanotubes with a diameter of 200 nm and the control group at day one(P>0.05).Conclusions Titania nanotubes with a diameter of 100 nm can improve the HGF attachment,proliferation,and secretion of typeⅠcollagen.
作者
许志强
曾秀峰
赖颖真
陈江
Xu Zhiqiang;Zeng Xiufeng;Lai Yingzhen;Chen Jiang(Department of Stomatology,Affiliated Hospital of Putian University,Putian 351100,China;Hanjiang Zeng Xiufeng Dental Clinic,Putian 351117,China;Department of Stomatology,Xiamen Medical College&Engineering Research Center of Fujian University of Stomatological Biomaterials,Xiamen 361023,China;Department of Oral Implantology,Affiliated Stomatological Hospital,Fujian Medical University,Fuzhou 350004,China)
出处
《中华口腔医学杂志》
CAS
CSCD
北大核心
2020年第4期253-258,共6页
Chinese Journal of Stomatology
基金
福建省自然科学基金(2017J01227)
福建省卫生计生中青年骨干人才培养项目(2017-ZQN-79)
莆田市科技计划(2018S3F007)。
关键词
钛
纳米管
成纤维细胞
细胞黏附
细胞增殖
Titanium
Nanotubes
Fibroblasts
Cell adhesion
Cell proliferation
作者简介
通信作者:陈江,Email:dentistjiangchen@sina.com,电话:0594-3793509。
