人牙周膜细胞在透明质酸钠/纳米羟磷灰石支架上的黏附与生长  

Adhesion and growth of human periodontal ligament cells on sodium hyaluronate/nanohydroxyapatite scaffolds

作　　者：张彦聪[1] 顾相伶[1] 刘洪玲[1] 付春华[1] 孙汉文[1] 

机构地区：[1]德州学院医药与护理学院,山东德州253023

出　　处：《山东医药》2013年第38期15-17,共3页Shandong Medical Journal

基　　金：山东省科学技术发展计划立项课题(2011YD02080);山东省自然科学基金资助项目(ZR2010BL001)

摘　　要：目的研究人牙周膜细胞(PDLC)在透明质酸(HA)/纳米羟磷灰石(nHAP)支架上的黏附、生长情况,以初步探讨HA/nHAP架应用于牙周组织工程的可行性。方法将体外培养的人PDLC分为HA/nHAP组、HA组、对照组,分别接种于加入了HA/nHAP支架、HA支架的24孔板及普通24孔板中,分别于1、2、3、4、5 d采用MTT法对接种的人PDLC进行光密度值(OD值)检测,倒置相差显微镜及免疫荧光染色后激光扫描共聚焦显微镜下观察细胞形态。结果第1、2、3天HA/nHAP组、HA组的OD值均高于对照组(P<0.05);HA/nHAP组高于HA组(P<0.05)。倒置相差显微镜下可见到各组孔板底部生长良好的梭形纤维细胞,核仁清楚可见,胞质均匀;HA/nHAP组支架材料完整,HA组支架材料大部分溶解。免疫荧光染色后激光扫描共聚焦显微镜下可见各组细胞核呈椭圆形,核完整,染色质均匀,未见核固缩和碎裂及异常核分裂。结论相对于HA支架,透明质酸钠支架有利于人牙周膜细胞的黏附生长,相对于透明质酸钠支架,HA/nHAP支架更有利于人牙周膜细胞的黏附生长,该材料有可能成为牙周组织工程的理想支架材料。Objective The adhesion and growth of human periodontal ligament cells （PDLCs） on the scaffolds of sodium hyaluronate （HA） and HA/nano-hydroxyapatite （nHAP） were studied to evaluate the feasibility of HA/nHAP scaffolds in periodontal tissue engineering.Methods Human PDLCs cultured in vitro were divided into the HA/nHAP group,HA group and the control group,which were respectively inoculated into the 24-pore plates of HA and HA/nHAP scaffolds and the normal 24-pore plate.The optical densities （OD） of inoculated PDLCs were detected by MTT assay on day 1,2,3,4 and 5.The cellular morphologies of PDLCs were observed through inverted phase contrast microscope and laser scanning confocal microscope after immunofluorescent staining.Results On day 1,2 and 3,the OD of HA and HA/nHAP groups was higher than that of the control group （P ＜ 0.05）,and HA/nHAP group was higher than HA group （P ＜ 0.05）.From the inverted phase contrast microscope,we saw the well-grown spindle cells with visible nucleoli and homogeneous cytoplasm at the bottom of each plate; HA/nHAP scaffold materials were complete,most of the HA scaffold materials dissolved.From laser scanning confocal microscope after immunofluorescent staining,we saw cells in different groups were oval and with complete nuclear and uniform chromatin,without karyopyknosis,fragmentation and abnormal mitosis.Conclusions HA/nHAP scaffold is more suitable for human PDLC adhesion compared with HA scaffold.This indicates that HA/nHAP scaffold might be a potential ideal candidate for periodontal tissue engineering.

关 键 词：牙周膜细胞 透明质酸 纳米羟磷灰石 支架材料 

分 类 号：R781.4[医药卫生—口腔医学]